Neuropsychiatric disorders represent the second largest cause of morbidity worldwide. These disorders have complex etiology and patho-physiology. The major lacunae in the biology of the psychiatric disorders include genomics, biomarkers and drug discovery, for the early detection of the disease, and have great application in the clinical management of disease. Indian psychiatrists and scientists played a significant role in filling the gaps. The present annotation provides in depth information related to research contributions on the molecular biology research in neuropsychiatric disorders in India. There is a great need for further research in this direction as to understand the genetic association of the neuropsychiatric disorders; molecular biology has a tremendous role to play. The alterations in gene expression are implicated in the pathogenesis of several neuropsychiatric disorders, including drug addiction and depression. The development of transgenic neuropsychiatric animal models is of great thrust areas. No studies from India in this direction. Biomarkers in neuropsychiatric disorders are of great help to the clinicians for the early diagnosis of the disorders. The studies related to gene-environment interactions, DNA instability, oxidative stress are less studied in neuropsychiatric disorders and making efforts in this direction will lead to pioneers in these areas of research in India. In conclusion, we provided an insight for future research direction in molecular understanding of neuropsychiatry disorders. Neuropsychiatric disorders represent the second largest cause of morbidity worldwide. These disorders have complex etiology. The genetic linkage is only 10% while remaining 90% are sporadic in nature. The World Health Organization has estimated that neuropsychiatric disease burden comprises 13% of all reported diseases. The psychiatric disorders include major depression, anxiety, schizophrenia, bipolar disorder, obsessive-compulsive disorder, alcohol and substance abuse, and attention-deficit hyperactivity disorder. Approximately one in five Americans experience an episode of a psychiatric illness such as schizophrenia, mood disorder (depression and bipolar disorder) or anxiety and a similar situation is predicted to be prevailing in developing countries too. The Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) defines the criteria for a wide array of mental illnesses. Most Psychiatrists use it as a basis for diagnosis. But diagnosis is still a challenge as many disorder episodes are overlapping.The prevalence of disorders, and their economics and societal aspects has to be understood through research programs aimed at elucidating the etiologies and pathophysiological mechanisms of these devastating disorders. The final goal will be clear diagnosis, management and drug discovery.Major depression is an affective disorder and the symptoms include feelings of profound sadness, worthlessness, despair and loss of interest in all pleasures. The individuals having depression also experience mental slowing, a loss of energy and an inability to make decisions or concentrate. The symptoms can range from mild to severe, and are often associated with anxiety and agitation. WorldwidBipolar disorder is a chronic mental illness, which begins, in adolescence or early adulthood. The typical bipolar disorders initiates between 15 and 25 years. The bipolar disorder onsets with initial episode of depression; these episodes of depression go as undiagnosed and are not treated appropriately. Bipolar Disorder includes Major Depressive Episodes and Hypomanic episodes. The significant portion of people suffering from manic depression did not have full manic episodes. The classification was divided into Bipolar I and Bipolar II. However, Bipolar II is often a first step to Bipolar I. It is seen that over five years, between 5 and 15% of those with Bipolar II change diagnosis to Bipolar I. Approximately 0.5% of people develop Bipolar II in their lifetimes. The psychobiology of therapeutic approach to anxiety disorders is still complex.). In China the prevalence was only half in India and, rural Latin America, a quarter or less of the European prevalence in rural India). The 10/66-dementia prevalence was higher than DSM-IV dementia, and more consistent across all sites varying between 5.6% (95% CI 4.2-7.0) in rural China and 11.7% (10.3- 13.1) in the Dominican Republic. The validity of the 847 of 1345 cases of 10/66 dementia not confirmed by DSM-IV was supported by high levels of associated disability (mean WHO Disability Assessment Schedule II score 33.7 [SD 28.6]). This indicates that the DSM-IV dementia criterion might underestimate dementia prevalence, especially in regions with low awareness of this emerging public-health problem.[60 in 11 et al.[There are a few reports on the clinical profiles of young-onset dementia from India. Nandi et al. conducteet al.[et al.[There is an international study linking a relationship between diet and dementia. Albanese et al. found thl.[et al. has contet al.[et al.[The amnestic and multiple domain types dementia are two types of dementia among nondemented and nondepressed elderly subjects aged 50 and older. This is a cross sectional community screening study. The subjects are selected by systematic random sampling for the assessment of cognitive function with the help of a validated cognitive questionnaire battery. The results indicated that prevalence of MCI detected based on neuropsychological testing is 14.89% (95% CI: 12.19 to 17.95). The prevalence of the amnestic type is 6.04% (95% CI: 4.40 to 8.1). The multiple domain type is 8.85% (95% CI: 6.81 to 11.32). The data insights that the amnestic type is more common among men and the multiple domain type among women with advancement of age. This is the first study on the prevalence of amnestic type in developing countries. Also, Das et al. has undeet al.[et al.[Rao et al. reviewedl.[et al. reportedet al.[Autism spectrum disorder (ASD) is a of complex neurodevelopment disorders, characterized by social impairments, communication difficulties. It is also characterized by abnormal behavior such as restricted, repetitive, and stereotyped behavioral patterns. The genetic imprinting of certain genes is reported in the autism. The paternal imprinting of HTR2A with expression from only one allele is reported. There are no reports on HTR2A and its association with neuropsychiatric disorders from Indian population. The study showed an association of the above-mentioned markers of HTR2A with Autism spectrum disorder (ASD) in Indian population. The genotyping analyses are carried out for probands, parents and controls. The results indicated that HTR2A is unlikely to be a genetic marker for ASD in Indian population. Recentlyet al. showed aet al.[Sexual dysfunction is a major risk factor for anxiety and depression. The reports of the prevalence of female sexual dysfunction (FSD) are scant. Recently, Singh et al. conducteet al.[et al.[Also, Kumar et al. examinedl.[et al. conducteErectile dysfunction (ED) is a complex condition where in men with minimal organic ED may develop a variable degree of psychogenic component sufficient to reduce the efficacy of medical management. Taneja studied To understand the genetic association of the neuropsychiatric disorders, molecular biology has a tremendous role to play. For example, to understand the inheritance pattern of the particular gene associated with the particular disorder, research in of molecular biology is essential. For example, the role of chromatin modulation in depression is new concept. The alterations in gene expression are implicated in the pathogenesis of several neuropsychiatric disorders, including drug addiction and depression. The development of transgenic neuropsychiatric animal models is a great thrust area. No studies from India in this direction. Biomarkers in neuropsychiatric disorders are of great help to the clinicians for the early diagnosis of the disorders. Gene-environment interactions, DNA instability, oxidative stress are less studied in neuropsychiatric disorders and making efforts in this direction will lead to pioneers in these areas of research in India. The molecular biology of the above mentioned areas would help to understand the neuropsychiatric disorders.In recent years, it has become widely recognized that a comprehensive understanding of chromatin biology is necessary to better appreciate its role in a wide range of diseases. The epigThere are fundamental contributions on genetics and gene-environment interactions in the field of neuropsychiatry from India. However, there are major lacunae in genomics, proteomics, biomarkers, drug discovery and development of animal models for neuropsychiatry disorders is a major thirst all over the word. The following are the major contributions from India.et al.[et al.[Genetics plays an important role in neuropsychiatric research and practice. It is important to understand the exact gene borne influences, mode of inheritance and establishing relationship between gene product and syndromes. Genetic markers like mini satellites, tandem repeat sequences to establish linkages are of great advantage. Rao et al. first col.[et al. studied et al.[LRRK2 gene among Indian PD patients, using PCR-RFLP method. They analyzed G2019S, R1441C, R1441G, and R1441H mutations in 800 PD and 212 controls, I2012T and I2020T mutations in 748 PD patients. LRRK2 gene encodes Leucine-rich repeat kinase 2, associates with the mitochondrial outer membrane. The authors did not find any of the above mutations, except in one female young onset PD patient who has a heterozygous G2019S mutation. They concluded that LRRK2 mutations may be a rare cause of PD among Indians. Dopaminergic pathway has been widely implicated in the pathophysiology of PD. Juyal et al.[Parkin gene is implicated in the PD, associated with mitochondrial dysfunction. Parkin gene encodes for Parkin protein (subunit of E3 ubiquitin ligase) involved in the proteosomal degradation. Chaudhary et al.[SNCA gene. Nagar et al.[et al.[et al.[P=0.028) marker, nominally significant genotypic associations for tyrosine hydroxylase rs6356 A/G (P=0.04) and dopamine beta-hydroxylase rs1108580 A/G (P=0.025) following the case-control approach. In dopamine beta-hydroxylase, catechol-O-methyltransferase, and dopamine receptor D (2) genes, several significant haplotypic associations were present. Gupta et al.[P-value = 0.0001).There are limited studies on the role of genes in neurological disorders. Punia et al. studied al et al. investigry et al. analyzedar et al. investigl.[et al. comparedl.[et al.–33 Srival.[et al. performeet al.[et al.[et al.[et al. reported a positive linkage and association for psychosis to the chromosomal position 18p11.2.[The study indicated that the interacting effects within the COMT gene polymorphisms may influence the disease status and response to risperidone in schizophrenia patients. Kumar et al. analyzedl.[et al. observedl.[et al. studied  18p11.2.et al.[et al.[Verma et al. identifil.[et al. also fouThere are several key issues that need to be resolved before we consider the clinical use of additional HDAC inhibitors to treat neuropsychiatric disorders such as schizophrenia or unipolar depression.. Recent et al.[The studies on candidate gene polymorphisms in a popula tion are useful for a variety of gene-disease association. A number of candidate genes from the monoaminergic pathway in the brain, have been associated with schizophrenia. In this study, diallelic/multiallelic polymorphisms in some dopaminergic, serotonergic and membrane-phospholipid-related genes have been evaluated in a control population recruited from North India. Eight genes tested association with schizophrenia for only two gene polymorphisms, one in the promoter region of the serotonin 2A receptor gene and the other in the tryptophan hydroxylase gene. One new allele for the dopamine transporter gene , has not been reported in any population. Ravi Kumet al. reportedThe concentration of trytophan, quinolinic acid, kynurenic acid, serotonin and 5-hydroxyindoleacetic acid was found to be higher in the plasma of patients with all these disorders; while that of tyrosine, dopamine, epinephrine and norepinephrine was lower. These observations are highly relevant in understanding the neuropsychiatric disorders.et al.[Fragile X syndrome (FRAXA), caused by the expansion of CGG repeats in the 5’ untranslated region of the fragile X mental retardation 1 (FMR1) gene is one of the most common forms of mental retardation. Due to the CCG repeats FMR1 gene becomes transcriptionally inactive by the hypermethylation. Guruju et al. performeet al.[et al.[et al.[Thelma et al. analyzedl.[et al. analyzedl.[et al. analyzedNeuronal cell death is observed in neurological and neuropsychiatric disorders. Several mechanisms and pathways were proposed for the neuronal cell death in these disorders. Gene-environment interactions play an important role in the regulation of neuronal cell death, here we focusing on the genes involved in neurological disorders. DJ-1 gene associated with early onset of neurological disorder Parkinson’s disease (PD) which coupled with dementia. DJ-1 belongs to a family protein which includes transcriptional regulators, proteases and chaperones.et al.[et al.[Saeed et al. showed tet al. A genderet al. Karunakal.[et al. showed cet al.[12 after removing the Al using a chelating drug desferoximine.Glutaredoxin helps in the recovery of complex 1 by regenerating the protein thiols. In the ret al. showed tet al.[P<0.02) in bipolar II hypomania, while in bipolar II depression, Na, K, Cu and Al are increased (P<0.001). Na, Mg, P, Cu, and Al are increased significantly (P<0.002) in bipolar V. In all 3 bipolar groups S (P<0.00001), Fe (P<0.002) and Zn (P<0.004) are decreased in all 3 bipolar groups.Interestingly, they found that even after the removal of Al (CCG)12 repeats remain in Z-DNA conformation. Probably the aluminum which is elevated in Fragile X syndrome people may alter FMR1 gene integrity and altered its expression levels. Mustak et al. estimateThe drugs used for the psychiatric disorders work differently among the patients. Genotype of the individuals affects the response of drugs differently. If genotype of the target molecules for the drugs is known it will be great advantage to the clinician to finalize type of the drug and dosage of the drug. Several people worked on the genotypes of patients for different psychiatric disorders in Indian populations. Cytochrome P450 (P450) enzyme plays an important role drug metabolism in the body. P450 enzymes also present brain and involved in the pharmacological modulation of drugs. Cytochrome P4502D constitutively expressed in the pyramidal neurons of CA1, CA2 and CA3 subfields of hippocampus, cerebral cortex, Purkinje and granule cell layers of cerebellum, reticular neurons of midbrain. Tissue-set al.[et al.[The above studies indicate that Cytochrome P4502D may play a role in the metabolism of psychoactive drugs directly at or near the site of action, in neurons, in human brain. Agarwal et al. (2008) sl.[et al. analyzedl.[et al. observedet al.[et al.[et al.[Tiwari et al. analyzedl.[et al. analyzedl.[et al. studied The major conclusions are:Neuropsychiatric burden is going to be in multifold in the next decadeThere is a great need to apply genetic knowledge base to predict and to manage the brain disordersWe need to discover biomarkers as early predictions of disease and as a endpoints for therapeutic interventionGene-environment interaction is necessary as an etiological feature to understand disease risk factorsMolecular genotyping for therapeutic intervention is the need of the hourLast, but the least, life style and diet as intervention events for diseases need to be exploited


Urinary tract infections (UTIs) are among the most common bacterial infections in humans, both in the community and hospital settings. It is a serious health problem affecting millions of people each year and is the leading cause of Gram-negative bacteremia. We previously conducted a study on “Urinary Bacterial Profile and Antibiotic Susceptibility Pattern of UTI among Pregnant Women in North West Ethiopia” but the study did not address risk factors associated with urinary tract infection so the aim of the study was to assess associated risk factors of UTI among pregnant women in Felege Hiwot Referral Hospital, Bahir Dar, North West Ethiopia.A total of 367 pregnant women with and without symptoms of urinary tract infection(UTI) were included as a study subject from January 2011 to April 2011. Midstream urine samples were collected and processed following standard bacteriological tests. Data concerning associated risk factors were collected using structured questionnaires and were processed and analyzed using Statistical Package for Social Science (SPSS version 16).Bivarait analysis of socio-demographic characteristics and associated risk factors of UTI showed that family income level (family monthly income level ≤ 500 birr($37.85); P = 0.006, OR = 5.581, CI = 1.658, 18.793 and 501–1000 birr ($37.93-$75.70), P = 0.039, OR = 3.429, CI = 1.065, 11.034), anaemia , sexual activity  and past history of UTI  were found to be factors significantly associated with increase prevalence of UTI. In contrast multiparity, history of catheterization, genitourinary abnormality, maternal age, gestational age and educational status were not significantly associated with UTI among pregnant women.In this study UTI was high among pregnant women in the presence of associated risk factor such as anaemia, low income level, past history of UTI and sexual activity. Urinary tract infection (UTI) is the single commonest bacterial infections of mankind, to help in alleviating the problem we previously conducted a study on UTI which addressed the bacterial profile and antibiotic susceptibility pattern of UTI ,2 but thPregnancy is one of the factors which increase the risk of UTI partly due to the pressure of gravid uterus on the ureters causing stasis of urine flow and is also attributed to the humoral and immunological changes during normal pregnancy . During Though there are few studies -15 conduThe methods were previously described in our study on “Urinary Bacterial Profile and Antibiotic Susceptibility Pattern of UTI among Pregnant Women in North West Ethiopia” except few differences .A hospital based cross sectional study was conducted at Felege Hiwot Referral Hospital (FHRH) from January 2011 to April 2011 to determine associated risk factors of urinary tract infection (UTI) among pregnant women.A pre-designed and structured questionnaire was used for the collection of data on associated risk factors. Collection of information on sign and symptoms of UTI and physical examination of pregnant women were done by Gynecologists and senior nurse.Pregnant women who were taken antibiotics within seven days at the time of recruitment and who were not willing to participate were excluded from this study.Clean catch mid-stream urine samples were collected using sterile, wide mouthed glass bottles with screw cap tops. The pregnant women were also informed to clean their hands with water and then cleanse their periurethral area with sterile cotton swab soaked in normal saline to reduce the risk of contamination. Urine specimens were processed in the laboratory within 2 hours of collection and specimens that were not processed within 2 hours were kept refrigerated at 4°C until it was processed.A calibrated sterile platinum wire loop was used for inoculation of specimens in to the culture media. It has a 4.0 mm diameter designed to deliver 0.01 ml. A loopful of well mixed urine sample was inoculated MacConkey, Manitol Salt Agar and Blood Agar .All plates were then incubated at 37°C aerobically for 24 hrs. The plates were then examined macroscopically for bacterial growth. The bacterial colonies were counted and multiplied by 100 to give an estimate of the number of bacteria present per milliliter of urine. A significant bacterial count was taken as any count equal to or in excess of 10,000 CFU/ml .Mid-stream urine specimen: - a specimen obtained from the middle part of urine flow: Clean catch urine specimen.Symptomatic UTI refers to patients whose urine is yielding positive cultures (≥ 105CFU/ml) and who have symptoms referable to the urinary tract.Asymptomatic bacteriuria (ASB) refers to the presence of two consecutive clear-voided urine specimens both yielding positive cultures (≥ 105CFU/ml) of the same uropathogen, in a patient without urinary symptoms.Maternal Anaemia is defined as haemoglobin concentration less than 11 g/dl.Parity is the number of pregnancy reaching viability or beyond stage of abortion (before 20 weeks/less than 500 g BW).Gestational Age is the age of the fetus estimated by computing from the first day of the last menstrual period (time that precedes conception) until the day of consultation.History of UTI is any history of infection pertaining to the urinary tract diagnosed by a physician.Pregnant women were informed to clean their hands with water and their genital area with swab soaked in normal saline before collection of the clean catch midstream urine samples. All specimens were transported from the hospital to regional laboratory within cold box and those specimens which were not processed with in 2hrs were kept in refrigerator and processed no longer than 18 hours after collection.5CFU/ml of urine were considered significant but specimen who produced < 105 colonies/ml of urine considered insignificant or due to contamination. Culture media were sterilized based on the manufactures instruction. Then the sterility of culture media were checked by incubating 3–5% of the batch at 35 – 37°C overnight and observed for bacterial growth. Those media which showed growth were discarded. The standard reference strains; Staphylococcus aureus (ATCC25923), Escherichia coli (ATCC25922) and P. aeruginosa (ATCC 27853) were used for testing quality of culture media.Only specimens which produced ≥ 10The study was conducted after getting a full approval by the health research unit of Jimma University, Amhara national regional state health bureau and FHRH. In addition written informed consent for the study was obtained from the study participants and confidentiality of results was kept. The results of urine tests were sent to the responsible person as soon as possible so that the pregnant women could be benefited from the study.A total of 367 pregnant women (37 symptomatic and 330 asymptomatic pregnant women) were investigated for the presence of risk factors associated with urinary tract infections.The assessment of associated risk factors of UTI showed that history of UTI , anaemia , sexual activity  and family monthly income (taking family monthly income > 2000 birr ($151.40) as reference, family monthly income level ≤ 500 ETB ($37.85); P = 0.006, OR = 5.581, CI = 1.658, 18.793 and 501–1000 ETB($37.85-75.70); P = 0.039, OR = 3.429, CI = 1.065, 11.034) showed statistical significant association with UTI.Bivariate analysis of other associated risk factors revealed that age of the pregnant women , gestational age , parity , history of catheterization , genitourinary abnormality  and educational status  were not significantly associated with UTI  was higher among pregnant women who had family monthly income of less than 500 Ethiopian birr ($37.85) and 501–1000 Ethiopian birr ($37.93-$75.70) for the overall UTI. Similarly study on the same study subject in Pakistan by Haider et al also sho al[et al on UTI a.9% was h al[et al.et al[et al[et al[et al[et al[Maternal anaemia was also significantly associated with UTI. Similar result were reported by different investigators such as study on UTI by Haider et al in Pakis al[et al among as al[et al among as al[et al among as al[et al in Thailet al[et al[et al[et al[et al[et al[The finding of this study also revealed that past history of UTI had strong association with UTI . Similar findings were reported by Haider  al[et al in Qatar al[et al also ide al[et al in Sudan al[et al in Thailet al[et al[et al[Sexual activity was also the other associated risk factor that was found to be significantly associated with UTI. Pregnant women who had recent sexual intercourse of three or more per week were more likely to have UTI than women who had less than three intercourses per week. This may be due to sexual activity increases the chances of bacterial contamination of female urethra. Having intercourse may also cause UTIs in women because bacteria can be pushed into the urethra . The anaet al on UTI i al[et al on UTI i al[et al on UTI aet al[et al[et al[et al[et al[et al[et al[Multiparity was associated with significant bacteriuria in pregnancy. This had been repeatedly recognized to cause a two-fold increase in the rate of ABU in pregnant women ,26. The et al on UTI i al[et al on UTI i al[et al among as al[et al among as al[et al on UTI i al[et al on asympet al[et al[et al[et al[et al[et al[et al[et al[There was no significant difference in the prevalence of UTI with respect to trimester. This is similar with earlier studies by Hamdan et al on UTI i al[et al on UTI i al[et al on UTI i al[et al in Iran, al[et al on asymp al[et al in Pakis al[et al on UTI i al[et al among as al[et al on UTI i al[et al on asympIn this study the chance of UTI was higher among pregnant women in the presence of associated risk factors such as anaemia, low income level, past history of UTI and sexual activity but there was no significant association between prevalence of UTI and risk factors such as multiparity, history of catheterization, genitourinary abnormality, maternal age, gestational age and educational status of pregnant women. Therefore pregnant women should be assessed for associated risk factors during their regular follow up.There is no additional supporting data; the supporting data is included as Additional file We verify that we all the authors have agreed to share the outcome of this manuscript equally and there is no conflict of interest among us.DT was responsible for selection of the topic and write up of the proposal and also responsible for the final write up. BG was responsible for designing the methodology of the study and also involved in the selection of the topic whereas TW was responsible for the analysis and interpretation of the data. MS was responsible for collection of samples and data. All authors read and approved the final manuscript.Questionnaire for socio-demographic, clinical data of symptomatic UTI and assessment of associated risk factors of UTI among pregnant women in FHRH.Click here for file


Molenberghs et al.  contribuMolenberghs et al.’s study is timely and much needed after the recent publication of new evidence concerning this debate (see, e.g., Saj et al., As the authors acknowledge in the general discussion, while their method based on “the peak coordinates of the critical lesion site” is certainly appropriate for gray matter lesions, it seems problematic for long-range white matter bundles. At variance with gray matter lesions, where one can look for maximum overlap (Vallar and Perani, At present, the only way to explore the possibility that a deficit results from disconnection of a particular fascicle is to track the relevant fascicle, draw the lesions, and see whether or not they are located along the fascicle (see, e.g., Bourgeois et al., Despite these caveats, Molenberghs et al.’s results did reveal that “the largest region involved in the development of spatial neglect was a white matter lesion corresponding to the superior longitudinal fasciculus”, consistent with the meta-analysis of Bartolomeo et al. . HoweverTo conclude, Molenberghs et al.  made an


Inversion of the Williams syndrome (WS) region on chromosome 7q11.23 has previously been shown to occur at a higher frequency in the transmitting parents of children with WS than in the general population, suggesting that it predisposes to the WS deletion. Frohnauer et al. recently reported that the frequency of this inversion is not elevated in the parents of children with WS in Germany relative to the German general population. We have compared Frohnauer et al.'s data to those from three previously published studies , all of which reported a significantly higher rate of 7q11.23 inversion in transmitting parents than in the general population. Results indicated that Frohnauer et al.'s data are consistent with previously reported frequencies of 7q11.23 inversion in North America and Spain in both transmitting parents and the general population. Dear Editor:agree with previously published reports of the frequency of inversion in Williams syndrome (WS) progenitors [not having a child with WS had one spouse with an inversion [We read with interest the article, "No significantly increased frequency of the inversion polymorphism at the WBS-critical region 7q11.23 in German parents of patients with Williams-Beuren syndrome as compared to a population control" -6. Frohngenitors -6 and ingenitors -6. The mFrohnauer et al. further report that in 5/24 couples who had a child with WS, one (4/24 couples) or both (1/24 couples) parents had an inversion [1 and Osborne et al. [Thus, in contrast to Frohnauer et al.'s  claim, te et al.  for a Noe et al.  and Norte et al. ,6 in the1Frohnauer et al. [r et al.  reportedWS: Williams syndromeThe authors declare that they have no competing interests.CAM, CBM and LRO conceived the study. CBM performed the statistical analysis. CAM, CBM and LRO drafted the manuscript. All authors read and approved the final manuscript.


Am J Trop Med Hyg 84:653–661 by Marzouki et al, there was an error in the published version of In Am J Trop Med Hyg 84:753–756 by Higazi et al, the fourth author's name is listed incorrectly. The correct name should be Wigdan A. Elmubark, not Wigdan A. Mohamed.In Am J Trop Med Hyg 84:851–857 by Inglis et al, the last author's middle initial was omitted. The author's name should be Christopher H. Heath.In Am J Trop Med Hyg 84:862–869 by Reyburn et al, an incorrect unit of measurement appears in the print version of the article. It should say “… an increase of 200 mm” in the sixth line of the abstract and on page 865 in the second column, line six.In


Monophasic fibrous synovial sarcoma (SS) is the most common variant of SS. Only a few cytological studies are available on this entity. Bcl-2 protein expression has been described as a characteristic marker of SS and is useful for its differentiation from other sarcomas. Cytokeratin and CD99 are also used in detecting SS.To evaluate synovial sarcoma and its variants cytomorphologically.During a period of 10 years 7 months, i.e. from January 1998 to July 2008, 12 cytologic specimens diagnosed as synovial sarcoma were reviewed. Ten cases were diagnosed as SS on aspiration alone but two cases required ancillary technique i.e., immunocytochemistry staining with bcl-2 and cytokeratin. The smears were stained with Papanicolaou and May-Grünwald-Giemsa stains.All cytologic specimens in our study had similar appearance. Most smears were highly cellular and were made up of densely packed tri-dimensional groups and singly scattered round to oval cells. Cellular monomorphism and vascular channels within the cell groups were the remarkable findings. Only one case showed cytologic evidence of epithelial differentiation. Bcl-2, cytokeratin, CD99 positivity was seen on immunohistochemistry staining. Results were categorized according to age, sex and morphologic variants.Although cytomorphologic features of synovial sarcomas are characteristic enough to permit its recognition, clinical correlation is necessary for accurate diagnosis. Monophasic variant is the most common entity observed in the present study. Synovial sarcoma (SS) is a mesenchymal spindle cell tumor which displays variable epithelial differentiation including glandular formation and has a specific chromosomal translocation t ×:18) (p11:q11)[8  using 23–25 gauge needle. Wet and air dried smears were prepared and stained subsequently with Papanicolaou and May-Grünwald-Giemsa stains, respectively. Ten cases were diagnosed on FNAC alone. Two cases required immunocytochemistry in view of the poor cellularity and staining for bcl-2 and cytokeratin were done. Of the total twelve cases, histopathologic correlation was available only for five cases. Histopathology ascertained the cytologic diagnosis in these five cases.Clinicopathologic features are summarized in Tables The monophasic sub type showed spindle cell sarcoma with hemangiopericytoma like foci and the biphasic type contained epithelial-like glands and nests. Presence of mast cells is also an important differential diagnostic feature of synovial sarcoma. Our results showed the same in three out of five cases. Depending on the availability of the markers, cytokeratin, S100 protein and CD34 were done. The S100 protein was done to exclude nerve sheath tumor. Cytokeratin and CD34 were positive and S100 protein was negative. The immunophenotype tends toward schwannian differentiation when S100 positivity is demonstrated (only in 50% of the cases), but is more commonly positive for LEU-7, collagen type 4 and myelin basic protein.All smears showed similar morphology. The smears were highly cellular with clusters and dispersed cell population. There were abundant single cells with naked nuclei. Cells were elongated, spindle to oval in shape and had uni and bipolar cytoplasmic processes. Nuclei were pleomorphic with hyperchromasia. There was presence of mast cells with infrequent mitoses and occasional calcifications. Myxoid matrix was seen stained purple to magenta color on May-Grünwald-Giemsa. Cell clusters were showing whorls and pericytic patterns in some and micro acini in a few cases . Clusteret al.[et al.[et al.[et al. showed female preponderance in the eleven cytologic specimens studied. The primary site is thigh in majority of the cases, which is similar with the present study [et al., but our experience shows that metastatic tumor were common after primary tumor. All cytologic specimens were obtained from fine needle aspiration by Viguer et al., except for two cases obtained from tumoral fluid. Present study does not show any evidence of cystic areas even in a single case, and shows monophasic fibrous variant as the most common on cytology and on histopathologic correlation as well [et al. stating that monophasic variant is the most common type of all morphologic variants of SS.FNAC plays a major role in diagnosing pre-operative mesenchymal lesions and distinguishes both benign and malignant soft tissue lesions. There are no major studies from India on synovial sarcoma alone emphasizing the role of FNAC in diagnosing the same except for a few case reports. Kumar et al. and Dey l.[et al. publishel.[et al.–5 Biphasl.[et al. A specifl.[et al.8 Many bil.[et al. Hence, il.[et al. In additl.[et al. Present l.[et al. Infrequel.[et al. in his snt study . Recurre as well . This stSS has to be differentiated from two broad based categories of lesions:Mesenchymal lesions with uniform spindle to round cell morphologyTumors with epithelioid cell morphologyBut the diagnostic difficulties are commonly encountered with hemangiopericytoma and fibrosarcoma.[et al., it cannot be considered critical in the cytologic diagnosis of synovial sarcoma. Bcl-2 protein shows intense cytoplasmic positivity but has to be differentiated from bcl-2 positive lymphomas.[osarcoma. Hemangioymphomas. Thirty pymphomas. Sarcomatymphomas. Cytogeneymphomas.et al.[Mohite et al. in theirIn the hands of experienced cytopathologists FNAC in conjunction with ancillary techniques has a diagnostic accuracy approaching 95% for the diagnosis of soft tissue sarcomas.Domanski stated tAlthough cytomorphologic features of synovial sarcoma are characteristic enough to permit its recognition, clinical correlation is necessary for its correct identification.


This review highlights our current understanding of metal-induced responses in plants, with focus on the production and detoxification of mitochondrial ROS. In addition, the potential involvement of retrograde signaling in these processes will be discussed.A general status of oxidative stress in plants caused by exposure to elevated metal concentrations in the environment coincides with a constraint on mitochondrial electron transport, which enhances ROS accumulation at the mitochondrial level. As mitochondria are suggested to be involved in redox signaling under environmental stress conditions, mitochondrial ROS can initiate a signaling cascade mediating the overall stress response,  Al. Al61]. d leaves . This su2O2 . A . A 65]. 2O2 ,71. Over2O2  and lackbidopsis . In durubidopsis ,74. As m . Fi. Fi2O2.   lacking functional complex I. This induces signaling throughout the cell to reset its antioxidative capacity completely, thereby coping with the loss of a major NADH sink and enhancing resistance to ozone and Tobacco mosaic virus [et al. [in vivo oxidation state of a redox-sensitive GFP targeted to Arabidopsis mitochondria. They demonstrated that mitochondria are highly sensitive to redox perturbation evoked by Cd, with their redox state recovering slower from an oxidative insult as compared to the cytosol or chloroplasts [Changes in mitochondrial electron transport and/or ROS production can have consequences for all other organelles in the plant cell. Indeed, plant mitochondria have a central position in the cellular carbon and nitrogen metabolism via the TCA cycle and their role in photorespiration . Dutilleet al.  have demic virus . Schwarz [et al.  studied roplasts . In addiroplasts ,33,77,90roplasts ,89. Howeet al. [Once ROS are formed in mitochondria of metal-stressed plants, they can either diffuse out of the mitochondria to mediate signaling functions or induce protein, lipid and DNA damage in the organelle itself  Figure . Compareet al.  in Al-stet al. .et al. [et al. [et al. [2O2 treatment. Mitochondrial aconitase seems particularly prone to oxidative damage [in vitro oxidation with Cu and H2O2 [Arabidopsis thaliana cells [Furthermore, plant mitochondrial proteins are susceptible to metal-catalyzed oxidation, leading to the irreversible formation of reactive carbonyl groups on amino acid side chains and hence reduced protein function . Substanet al. . Bartoli [et al.  have dem [et al. , who have damage  as it isand H2O2  or fragmand H2O2 . In addina cells .2O2 and other ROS as signals modulating plant PCD. The main event in plant PCD is the release of mitochondrial cytochrome c [et al. [Arabidopsis mesophyll protoplasts using fluorescence techniques to monitor the in vivo behavior of plant mitochondria and caspase-3-like activity. After a quick ROS burst, mitochondrial swelling and loss of the transmembrane potential occurred prior to PCD. Application of AsA prior to Al-exposure slowed down but did not prevent these processes [2°− production—rather than NADPH oxidase-derived extracellular H2O2—was shown to be a key event in Cd-induced cell death in tobacco cells [et al. [et al. [Although enhanced mitochondrial ROS levels may serve as monitors and signal the extent of environmental stress throughout plant cells, they may also lead to oxidative damage and programmed cell death (PCD) when mitochondrial and/or cellular antioxidative defense and repair systems are overwhelmed. Programmed cell death is an active and genetically controlled process essential for growth and development, as well as for adaptation to altered environmental conditions . In animchrome c . This wa [et al. . In addi [et al. . Recentl [et al.  investigrocesses . Mitochoco cells . In addi [et al.  and rece [et al. . Althouget al. [et al. [et al. [Depending on the intensity of the stressor, metals are able to induce mitochondrial damage and/or signaling outside the mitochondria. An altered organellar redox state generates signals that are transmitted to the nucleus in a process called retrograde signaling. This process occurs between mitochondria, chloroplasts and the nucleus and can be mediated by ROS or oxidative stress-induced secondary signals. Recently, Suzuki et al.  reviewed [et al.  summariz [et al. . They su [et al.  reviewed [et al. ,103.AOX transcription is increased [et al. [To date, no specific components of any mitochondrial retrograde signaling pathway have been identified . Howeverncreased ,104. The [et al.  demonstrTobacco mosaic virus in tobacco plants [et al. [Arabidopsis resulted in acute sensitivity to combined drought and light stress, confirming a role for AOX in determining the steady-state cellular redox balance [Arabidopsis protoplasts lacking the gene coding for the major isoform AOX1a showed a dramatically decreased viability during Al exposure as compared to wildtype protoplasts [AOX1a overexpression enhanced Al tolerance, confirming the protective role of AOX against Al-mediated PCD [2O2, salicylic acid and the protein phosphatase inhibitor cantharidin. It is hypothesized that the survival function of AOX is based on its ability to continuously suppress mitochondrial ROS generation. This further prevents oxidative damage that could otherwise evoke disturbed gene expression and favor PCD. In addition, the maintenance of respiration in stressful conditions by the alternative route also contributes to the hypothesis of AOX as a survival protein [et al. [The most intensively studied model for retrograde signaling between the mitochondrion and nucleus resulting in acclimation to stress conditions is AOX  ROS in metal-induced PCD has been confirmed in several studies. Functioning as both a target and regulator of stress responses in plants, AOX is of major importance in the mitochondrial metabolism. Due to its ability to reduce mitochondrial ROS production, modulate PCD and TCA cycle activity, AOX is suggested to play a key role in metal-induced responses in plant mitochondria . Further


Systemic autoimmune diseases are a broad range of related diseases characterized by dysregulation of immune system which give rise to activation of immune cells to attack autoantigens and resulted in inappropriate inflammation and multitissue damages. They are a fascinating but poorly understood group of diseases, ranging from the commonly seen rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) to the relatively rare systemic sclerosis . The mecBased on this background, we assembled this special issue for a better understanding of systemic autoimmune diseases, on aspects of mechanisms of pathogenesis, diagnosis, and treatment, including papers ranging from the basic researches to clinical researches and reviews about systemic autoimmune diseases.β2-AR) and p53 apoptosis effector related to PMP-22(Perp) in the pathogenesis of RA, respectively. It has long been demonstrated that γδ T cells play important roles in the development of autoimmune diseases; the precise role of γδ T cells in the pathogenesis of SLE was studied by Z. Lu et al. Z. Gu et al. discussed the role of p53/p21 pathway in the pathogenesis of SLE. X. Gan et al. demonstrated the role of GITR and GITRL in the primary Sjögren's syndrome. The expression of IL-6 and its clinical significance in patients with dermatomyositis was discussed by M. Yang et al. L. Estrada-Capetillo et al. found that DCs from patients with rheumatic inflammatory disease show an aberrant function that may have an important role in the pathogenesis. S. Stratakis et al. studied the mechanisms underlying this beneficial effect of rapamycin in passive and active Heymann nephritis (HN).Studies on the basic research of systemic autoimmune disease in this issue provided us new insights into the mechanism of the pathogenesis of systemic autoimmune disease. The role of HMGB1 in the T-cell DNA demethylation was discussed in the paper of Y. Li et al. SNPs with strong RA association signal in the British were analyzed in Han Chinese by H. Li et al., and the methylation status of miR-124a loci in synovial tissues of RA patients was analyzed by Q. Zhou et al. indicating the epigenetic factor in the pathogenesis of RA. Expression of microRNA-155 was studied by L. Long et al. in RA patients. IL-33 status was tested in RA patients by S. Tang et al. D. Lorton et al. and Y. Du et al. indicated the possible role of beta2-adrenergic receptors  and pulmonary function test (PFT) abnormalities capable of identifying asymptomatic, preclinical RA-ILD. L. Pan et al. made a retrospective study to compare the characteristics of connective tissue disease-associated interstitial lung diseases, undifferentiated connective tissue disease-associated interstitial lung diseases, and idiopathic pulmonary fibrosis. Relationship between Brachial-ankle pulse wave velocity (baPWV) and its associated risk factors in Chinese patients with RA was analyzed by P. Li et al. P. Žigon et al. studied the diagnostic value of antiphosphatidylserine/prothrombin antibodies in systemic autoimmune disease. The correlations of disease activity, socioeconomic status, quality of life, and depression/anxiety in Chinese SLE patients were studied by B. Shen et al. J. Li et al. made a systematic review on efficacy and safety of Iguratimod for the treatment of rheumatoid arthritis.γR-mediated trogocytosis in the physiological immune system was discussed by S. Masuda et al.Review papers also cover many aspects about systemic autoimmune disease. Advances in the knowledge of costimulatory pathways and their role in SLE were discussed by N. Y. Kow and A. Mak H. Draborg et al. summed up existing data about the relationship between epstein-barr virus and autoimmune disease. T. Marchetti et al. discussed obstetrical antiphospholipid syndrome from pathogenesis to the clinical and therapeutic implications. Y. f. Huang et al. summarized the immune factors involved in the pathogenesis, diagnosis, and treatment of Sjogren's syndrome. The role of IL-33 in rheumatic diseases was reviewed by L. Duan et al. T. Ito made a review on advances in the pathogenesis of autoimmune hair loss disease alopecia areata. A. W. J. M. Glaudemans reviewed the use of 18F-FDG-PET/CT for diagnosis and treatment monitoring of inflammatory and infectious diseases. The role of FcThis special issue covers many important aspects in the systemic autoimmune diseases ranging from novel insights into the pathogenesis of autoimmune disease and the use of newly developed diagnostic strategy in the early diagnosis of autoimmune disease to the treatment of these kinds of diseases, which will surely provide us a better understanding about systemic autoimmune disease.Guixiu  ShiJianying  ZhangZhixin  (Jason)  ZhangXuan  Zhang


The aim of microfluidic mixing is to achieve a thorough and rapid mixing of multiple samples in microscale devices. In such devices, sample mixing is essentially achieved by enhancing the diffusion effect between the different species flows. Broadly speaking, microfluidic mixing schemes can be categorized as either “active”, where an external energy force is applied to perturb the sample species, or “passive”, where the contact area and contact time of the species samples are increased through specially-designed microchannel configurations. Many mixers have been proposed to facilitate this task over the past 10 years. Accordingly, this paper commences by providing a high level overview of the field of microfluidic mixing devices before describing some of the more significant proposals for active and passive mixers. For example, the Reynolds number is of the order of 0.1 in a typical water-based microfluidic system with a channel width of 100 μm, a liquid flow rate of 1 mm/s, a fluid density of 1 g/cm3 and a viscosity of 0.001 Ns/m2. In such low Reynolds number regimes, turbulent mixing does not occur, and hence diffusive species mixing plays an important role but is an inherently slow process. Consequently, the aim of microfluidic mixing schemes is to enhance the mixing efficiency such that a thorough mixing performance can be achieved within shorter mixing channels, which can reduce the characteristic size of microfluidic devices. Furthermore, the development of efficient mixing schemes is essential for increasing the throughput of microfluidic systems and to realize the concept of micro-total-analysis systems and lab-on-a-chip systems.Microfluidic devices have had a considerable impact on the fields of biomedical diagnostics and drug development, and are extensively applied in the food and chemical industries. The diminutive scale of the flow channels in microfluidic systems increases the surface to volume ratio, and is therefore advantageous for many applications. However, the specific Reynolds number  of the multiple species, active mixing schemes improve the mixing performance by applying external forces to the sample flows to accelerate the diffusion process . TypicalThe microfluidic mixers presented above are all considered for the mixing of continuous bulk liquids. However, various discrete droplet-based mixing platforms have also been proposed. One such scheme involves the use of air pressure to form, actuate and mix two liquid droplets in a hydrophobic microcapillary valve device. Recently, many studies have presented mixing devices using liquid droplets based on electro-wetting phenomena. In these schemes, electro-wetting actuation is applied to separate liquid droplets from the bulk and to drive them to specific locations where they are repeatedly combined, mixed and separated. The microfluidic mixing of liquid droplets through the application of electro-wetting-induced droplet oscillations has also been demonstrated. These microfluidic mixing schemes take advantage of the open structure of the flow channel, allowing the mixed sample to be more easily transported to its required destination.2.Active microfluidic mixers enhance the mixing performance by stirring or agitating the fluid flow using some form of external energy supply. As shown in 2.1.et al. [et al. [et al. [et al. [et al. [Liu et al.  demonstr [et al.  employed [et al.  presente [et al.  proposed [et al.  presente2.2.et al. [When the dielectrophoretic (DEP) force is applied to mixing, non-uniform alternating electrical fields induce the motion of polarized particles/cells . The eleet al.  proposed2.3.et al. [Electrokinetic time-pulsed microfluidic mixers apply an electrokinetic driving force to transport the sample fluids while simultaneously inducing periodic perturbations  in the fet al.  proposed2.4.In pressure perturbation mixers, perturbations within the fluid streams are generated by velocity pulsing . In a ty2.5.et al. [In the micromixer presented by El Moctar et al. , two flu2.6.et al. [Tsai et al.  presenteet al. [The intrinsically oscillatory flow generated by the bubble actuated nozzle-diffuser micropump was shown to induce a wavy liquid interface, accelerating the mixing process. Xu et al.  showed t2.7.et al. [et al. [T between one of the wire electrodes and the circular side-wall electrode and then between the second wire electrode and the side-wall electrode. Particle tracing revealed that chaotic flows were induced and resulted in a satisfactory mixing result within 40 periods. Wang et al. [The magneto-hydrodynamic (MHD) flow effect has been used by various researchers to realize micromixers. For example, Bau et al.  develope [et al. . The devg et al.  develope2.8.et al. [et al. in [The use of electrokinetic instability (EKI) as a mixing technique for electrokinetically-driven microfluidic flows with conductivity gradients has received considerable attention in recent years ,47. In aet al.  developet al. in . However3.Passive micromixers contain no moving parts and require no energy input other than the pressure head used to drive the fluid flows at a constant rate. Due to the inherently laminar characteristics of micro-scaled flows, mixing in passive micromixers relies predominantly on chaotic advection effects realized by manipulating the laminar flow within the microchannels or by enhancing molecular diffusion by increasing the contact area and contact time between the different mixing species. 3.1.et al. [et al. [et al. [et al. [In miniaturized flow systems with Reynolds numbers varying from 2 to 100, flow structures can be artificially induced, assisting flow segmentation through inertia effects. In the zigzag channel considered in , segmentet al.  who fabr [et al.  presente [et al.  proposed [et al.  fabricat3.2.et al. [et al. [Micromixers with intersecting channels can be used to split, rearrange and combine component streams to enhance mixing . He et aet al.  proposed [et al.  presente3.3.et al. [s on the mixing efficiency was investigated in a series of experimental trials. The results indicated that for Re = 0.26, the mixing efficiency increased from 65% to 83.8% as the geometry ratio s/w was increased from 1 to 8. For low values of s/w, the number of angles increased, resulting in an increase in the effective width and a reduction in the effective length. For low Reynolds number flows, the most efficient zigzag configuration corresponding to s = 800 μm obtained a mixing efficiency of 83.8%. For Re = 267, the mixing efficiency increased rapidly to 98.6% as the geometry ratio was increased to 4, but reduced slightly to 88.1% as the geometry ratio was further increased, thus indicating the existence of an optimal zigzag geometry. In [et al. presented a passive micromixer with a modified Tesla structure. In the proposed design, the species streams flowed close to the angled surface due to the Coanda effect, and this effect was used to guide the fluid streams to collide with one another. Mixing cells in opposite directions were then used to repeat the transverse dispersion caused by the flow impact. In the micromixer, one of the fluids was divided into two sub-streams and one of these two sub-streams was then merged with the second fluid stream from the main channel in the micromixer. The two streams were then mixed with the second sub-stream, resulting in a strong impact around the sub-channel of the micromixer. The results showed that the micromixer attained an excellent mixing performance at higher flow rates, and was characterized by a pressure drop of less than 10 kPa for flow rates of approximately 10 μL min−1. However, at lower flow rates, the mixer was constrained to the diffusive mixing regime, and hence the mixing performance was limited.Mengeaud et al.  presente3.4.et al. [et al. [et al. [Re = 1, the mixing performance of both mixers varied inversely with the mass fraction of the sample due to the dominance of molecular diffusion. However, when the Reynolds number was increased to 10, the inverse trend was observed in the serpentine mixer. This phenomenon was attributed to an enhanced flow advection effect at large sample mass fractions. However, this effect was not observed in the herringbone mixer when the Reynolds number was increased over a similar range. Liu et al. [et al. [et al. [i.e., sense of rotation of two rotational flows, aspect ratio of channel and ratio of bypass channel to whole width) and found at proper combination of the variables, almost global chaotic mixing was observed in the Stoke flow regime. Moon et al. [Stroock et al.  proposed [et al.  presente [et al.  consideru et al.  fabricat [et al.  investig [et al.  simulaten et al.  presente3.5.et al. [et al. [Re = 1.24 × 104 with a Rhodamine diffusivity of 2.8 × 10−10 m2s−1). Laser scanning of the entrance zone of the micromixer identified a bright image only in the half-zone containing Rhodamine. Bright images were also observed at the no-barrier zone in the first half-cycle, thus confirming the cross-sectional rotating flow effect induced by the slanted grooves. When the streams entered the barrier zone in the next half-cycle, laser scanning showed that the flow had rotated yet further. The experimental results confirmed that the barrier embedded micromixer yielded excellent species mixing within a short length of channel. Recently, Singh et al. [xN, the number of parallel cross-bars per element, pN, and the angle between opposite cross-bars. An optimum series for all possible SMX(n) designs to obey the universal design rule is pN = (2/3) xN − 1, for xN = 3, 6, 9, 12…Keoschkerjan et al.  fabricat [et al. ,73 develh et al.  analyzed3.6.et al. [et al. [p and depth ratio α of the groove on the mixing performance of a staggered herringbone mixer with patterned grooves. Because the two vortices within the mixing channel are determined by the asymmetry index p, vortices with dissimilar scales were shown to provide a better mixing performance than two equally-sized vortices. Furthermore, the results showed that the intensity of the vertical fluid motions at the side edges of the grooves increased with increasing groove depth α, resulting in a significant improvement in the mixing effect.Johnson et al. ,75 prese [et al.  investig3.7.et al. [et al. [et al. [et al. [Jen et al.  presente [et al.  presente [et al.  presente [et al.  presente3.8.−). When these surfaces come into contact with a solution containing ions, the positive ions are attracted to the surface, forming an important diffuse layer [In microfluidic systems, very high pressure gradients are generally required to drive and manipulate the fluid flow. Due to the small characteristic scale of the microchannels in typical microfluidic devices, surface forces dominate, and high friction effects are generated. Conventionally, microchannels are fabricated using silicon dioxide. Silicon dioxide surfaces are typically negatively charged due to their deprotonated silanol groups (≡Si–Ose layer . If an ese layer . Mixing se layer . By selese layer . In nume4.Advances in MEMS techniques in recent decades have enabled the fabrication of sophisticated biochips for a diverse range of applications. Compared to their traditional macro-scale counterparts, micromixers have a shorter operation time, a lower cost, an improved portability and a more straightforward integration with other planar bio-medical chips. This paper has presented a systematic review of the major micro-mixers presented in the literature over the past 20 years or so. It has been shown that depending on their mode of operation, these micromixers can be broadly categorized as either active or passive. The operational principles and mixing performance of each type of micromixer have been systematically discussed, and their relative advantages and disadvantages highlighted where appropriate. Overall, the results presented in this review confirm the applicability of micromixers for a diverse range of low-cost, high-performance microfluidic applications.


In addition, a portable system for monitoring salivary α-amylase activity was launched in Japan at the end of 2005. The correlation between exercise and salivary α-amylase has since been extensively investigated. The present review summarizes relevant studies published in the English and Japanese literature after 2006. A search of the PubMed and CiNii databases identified 54 articles, from which 15 original articles were selected. The findings described in these publications indicate that exercise consistently increases mean salivary α-amylase activities and concentrations, particularly at an intensity of >70% VO2max in healthy young individuals. Thus, these studies have confirmed that salivary α-amylase levels markedly increase in response to physical stress. Salivary α-amylase levels may therefore serve as an effective indicator in the non-invasive assessment of physical stress.The secretion of salivary α-amylase is influenced by adrenergic regulation of the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis; thus, exercise affects the levels of salivary α-amylase. Granger  Salivary α-amylase secretion is influenced by adrenergic regulation of the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis . Therefoet al  and CiNii (http://ci.nii.ac.jp/) databases. The latter is a database maintained by the Japanese National Institute of Informatics , which comprises literature published by Japanese authors in academic journals or university memoirs and is listed in the database of the Japanese National Diet Library .Information was collected from the PubMed  or peak power output of the study participants and four used ergometers (Eight studies defined exercise intensity as a ratio (%) of the maximum or peak oxygen uptake . Allgrove et al . Leicht et al (et al (2max for 2 h. The findings of Rosa et al (2max for 1 h supported these results; the mean salivary α-amylase concentrations were increased but the increase was not statistically significant. Three of the four studies, with the exception of the study of wheelchair athletes, comprised small cohorts, which may account for this discrepancy.By contrast, treadmill running generated mixed results. Fortes and Whitham  observedht et al  reportedl (et al , salivarsa et al  from a set al (et al (et al (et al (Five studies demonstrated changes in salivary α-amylase in response to exercise without specifying the exercise intensity –18. In oet al  adopted et al . Salivaret al . Allgrovl (et al  examinedl (et al  observedl (et al  and relal (et al –16.et al (et al (et al (Chiodo et al  and Diazl (et al  investigl (et al . Diaz etl (et al  compared2max in healthy young individuals. Therefore, studies published following those reviewed by Granger et al (In conclusion, exercise has consistently been shown to increase mean salivary α-amylase activity and concentration in all studies examined in the present review, including those in which changes were not significant, with the exception of the 20-min forest walk . The effer et al  confirm


There has been much interest in the mechanisms by which calcium may attenuate weight gain or accelerate body fat loss. This review focuses on postprandial energy metabolism and indicates that dietary calcium increases whole body fat oxidation after single and multiple meals. There is, as yet, no conclusive evidence for a greater diet induced thermogenesis, an increased lipolysis or suppression of key lipogenic enzyme systems. There is however convincing evidence that higher calcium intakes promote a modest energy loss through increased fecal fat excretion. Overall, there is a role for dietary calcium in human energy metabolism. Future studies need to define threshold intakes for metabolic and gastrointestinal outcomes. Body weight and body energy content remains quite stable in most adults for long periods of time; despite daily fluctuations in energy intake and energy expenditure. This requires the presence of regulatory processes able to match fuel supply to energy requirements. The maintenance of a physiological set point for body weight is complex and includes aminostatic and glucostatic controls of feeding, metabolic or nutrient partitioning, input from the sympathetic nervous system, signals from adipose tissue as well as additional behavioral influences. Genetics, the environment and psychosocial factors also impact on these regulatory processes ,2,3. Thaet al. [2+) held the key to fat deposition and obesity. According to this early scheme —a key enzyme regulating lipid deposition—while stimulating adipose tissue lipolysis. An increased fat oxidation and thermogenesis through up-regulation of uncoupling proteins (UCP) was also suggested. A role for calcium in the regulation of body weight has gained much interest since the early observation that increased intakes of calcium (as yoghurt) increased the loss of body fat and favoured a redistribution of fat away from the abdomen . Based pet al.  proposedy scheme  increaseWe now know that dietary calcium may also act at the level of the gastrointestinal tract to increase energy loss, through increased fecal fat excretion . TordoffIn the main, energy balance is determined by the matching of macronutrient intake to energy expenditure and the ability to channel nutrients into oxidative versus storage pathways (nutrient partitioning). Most individuals reach a state of approximate weight maintenance in which the average composition of the fuels they oxidize matches the nutrient distribution in their diets . Acute iThere are three main contributors to total energy expenditure (TEE) of man: basal metabolic rate (BMR) has the largest energetic demand accounting for 60–70% of TEE in sedentary individuals, diet induced thermogenesis (DIT) is a variable 10% and physical activity makes up the remainder. BMR is determined by fat free mass (FFM) to a great extent (80–85%) and by fat mass (FM) if the person is obese. Smaller contributions are made by age, habitual physical activity and genetics. DIT is traditionally calculated as the change in postprandial energy expenditure from fasting and expressed as a percentage of the energy content of the meal. DIT has two components: an obligatory process of energy expenditure that is strongly related to meal size, composition and the physiological characteristics of the individual, and a regulatory component that is strongly influenced by the SNS. A higher DIT for a given meal would imply that less energy is available for storage, and hence such a meal would be less conducive to weight gain. Abdominal adipocytes are particularly sensitive to SNS mediated lipolysis; a process whereby stored fat is broken down and mobilized. This paper focuses on the potential postprandial events that would contribute to an anti-obesity effect of calcium based on the model in et al. [Some methodological considerations are pertinent to the overall discussion on calcium and postprandial events. It is accepted that the previous evening’s meal composition affects the next day’s glycaemic response and macronutrient oxidation . Since pet al.  observedet al. ,18, noneThe absorption of calcium is not straight forward and is highly variable between subjects. There are many factors that may confound its absorption . After set al. [vs. 243 mg) modified postprandial FOR, DIT, FFA and glycerol with carry over effects to a standard lunch, low in D calcium (48 mg). Overall, there was a greater DIT and FOR after the higher breakfast-lunch trial [The rise in insulin in response to mixed meal ingestion drives thermogenesis and promotes an increased carbohydrate oxidation rate (COR), with a suppression of fat oxidation rate (FOR). Circulating free fatty acids (FFA) and glycerol are also suppressed. Cummings et al.  tested wet al. ,27. The et al. ,27,28. Wet al. ,29. We och trial ,29. Thisch trial . Togetheet al. [i.e., rest, sleep and exercise. Similarly postprandial 24 h FOR was not statistically different between treatments but a clear rank order (D > ND > control) was evident with a difference of ~8 g/d between high D and control diets. The authors calculated that such a difference could account for a loss of ~3 kg body weight if extrapolated to a year. Interestingly these trends for a greater 24 h FOR were paralleled by a similar trend for a greater negative fat balance across trials. In fact we note that in this study there was a significant negative fat balance following high D  but not after the other two. The study utilized high protein diets from either dairy or non-dairy sources, and also provided more dietary fibre than habitual consumed by the participants. Both components would be expected to influence calcium absorption and retention [There are four crossover trials that manipulated calcium intake over a week and measured postprandial DIT and FOR . Boon etet al.  achievedetention ,32, but et al. [vs. 23%E) for a week, and compared them to a control diet . Whole body calorimetry (WBC) measurements at the start and end enabled the calculation of change over that week. There was a small but significant drop in weight across all 3 diets; accompanied by a significant drop in RQ. However, there were no differences in 24 h EE, energy balance or FOR. A closer examination of their results indicated that the change in 24 h COR adjusted for energy balance, favoured a greater suppression on the two high calcium diets. Reciprocally, the change in 24 h FOR showed a greater increase (~ 35%) on the high D (15%E protein) diet relative to control [Using a similar design, Jacobsen et al.  varied D control . The sma control . In anot control . On the et al. [St-Onge et al.  studied Ten obese men and women with calcium intakes <800 mg/d completed a double blind placebo controlled crossover study of 5 weeks. Calcium was provided as a milk mineral sachet twice a day to increase calcium by ~800 mg/d, over and above habitual intake. The control group received a placebo . This elegant study used tracer isotope technology, indirect calorimetry, micro-dialysis of sub-cutaneous fat and fat biopsies for gene expression. The authors reported no statistical difference in DIT or FOR over the 6h postprandial period . In this3 and change in DIT . The relationship of DIT to vitamin D status at baseline is interesting. It suggests that the ability of calcium to stimulate DIT is dependent on its absorption; a function of vitamin D status [3 is linked to thermogenesis either directly or via PTH. Overall, ND calcium decreased RQ at rest and following a meal. In a randomized parallel design, 3 groups of overweight and obese individuals were assigned to either control (~500 mg/d from 1 serve D plus placebo), high calcium  and high calcium  during 12 weeks of energy restriction . This was a sub-study of a multisite trial on the source of calcium and energy metabolism . There wD status . This maD status . An alteThere was only one long term randomized parallel study conducted over a year . In this3 status and change in FOR [et al. [et al. [Dairy sources contain other bioactive components besides calcium and may be expected to show a greater effect. Dairy naturally contains vitamin D, which may be significantly higher in those countries that have mandatory fortification of their products. While vitamin D will enhance calcium absorption, there is some suggestion of an independent effect from the direct relation between 25(OH)De in FOR  and the e in FOR . Furthere in FOR ,35 and be in FOR . A simil [et al. , however [et al.  found thet al. [et al. [et al. [In summary this analysis shows that in 6 of 9 randomized studies that measured macronutrient oxidation, an increase in FOR following calcium was demonstrated . We coulet al.  for reas [et al.  and Jaco [et al.  which shet al. [et al. [et al. [One expected effect of higher intakes of dietary calcium, is the reciprocal stimulation of lipolysis and suppression of de novo lipogenesis . Coppacket al.  and Frayet al.  have arget al. ,18,30. W [et al. ,40 are o [et al.  observed [et al. . A role  [et al. . Recent  [et al. . No diffet al. [et al. [et al. [We found only 3 human studies that have measured the mRNA expression of genes regulating fat metabolism, of which two were conducted by the same group. In the earlier study of Boon et al. , D and N [et al.  tested a [et al.  studied Fecal fat excretion stems from exogenous (dietary) and endogenous sources . Further, unabsorbed fat may be metabolised by bacteria in lower GI tract and hence not be excreted . While net al. [et al. [In et al.  found a  [et al.  favoured [et al. . et al. (44) estimate that additional calcium may account for between 2–5 g/d greater loss of fecal fat. The effect of D calcium was more consistent than ND calcium, in part due to the many formulations of ND used. A regression analysis on a small number of subjects, predicted that ~1241 mg/d of D calcium would account for ~5.2 g/d of fat excretion. The authors estimate that such effects at best, would account for a 2.2 kg difference in body weight over 1 year [The study of calcium’s effect on fecal fat excretion predates the postulation of the calcium-body weight hypothesis, and there are many older studies that document increases in fecal fat excretion following higher calcium ,42,43. Aet al.  estimater 1 year ,46 but iIt is our opinion that calcium modulates human energy metabolism. There is convincing evidence that calcium increases postprandial fat oxidation after a single meal or over a day. There is some suggestion that fasting FOR may be greater on a high calcium diet. However a higher DIT is not always observed. There is convincing evidence for calcium to modestly increase the amount of fat excreted, and thereby contribute to energy loss. The effect of dietary calcium on lipolysis and/or lipogenesis is inconsistent at present, and is a fertile area for future research. Nutrition and public health recommendations demand consistent observations with a strong mechanistic basis. Defining threshold effects for metabolic versus gastrointestinal function based on randomized controlled trials with adequate power, is one way forward.


Cell migration and invasion are processes that offer rich targets for intervention in key physiologic and pathologic phenomena such as wound healing and cancer metastasis. With the advent of high-throughput and high content imaging systems, there has been a movement towards the use of physiologically relevant cell-based assays earlier in the testing paradigm. This allows more effective identification of lead compounds and recognition of undesirable effects sooner in the drug discovery screening process. This article will review the effective use of several principle formats for studying cell motility: scratch assays, transmembrane assays, microfluidic devices and cell exclusion zone assays. It isThe migration process in wound healing is coordinated among several different cell types including keratinocytes, fibroblasts, endothelial cells and macrophages that provide a variety of growth factors. Platelet derived growth factor (PDGF-BB), basic fibroblast growth factor (bFGF) and granulocyte macrophage colony stimulating factor (GM-CSF) are used as stimulants to facilitate wound healing for patients with diabetic ulcers . These gin vitro [Similar to the complex mechanisms underlying the process of wound healing, many different factors are also involved in cancer cell metastasis. There are diverse mechanisms that cells can employ to initiate and progress invasion and each offers specific pharmacologic targets for development of anti-metastatic therapies . In ordein vitro . TherefoThe goal of HTS is to accelerate the drug discovery process by rapidly evaluating large compound libraries . To achiet al. [in vivo-like data for all segments of the drug discovery pipeline, such as target validation, screening and toxicology.” Similarly, Carragher [in vitro assays that employ 3-dimensional matrices to provide relevant microenvironments for cellular studies [in vivo mechanisms of invasion [Justice et al.  assessedarragher  points o studies ,10. Inde studies . As a coinvasion . Such moinvasion ,19.In this article, we review the different assay formats employed to study cell motility. For many years, Boyden chamber based transmembrane assays and scratch wound assays were the only widely available formats to study cell migration and invasion. However, new technologies such as microfluidic chambers and exclusion zone assays have recently emerged as alternative phenotypic screening assays that provide additional or complementary information to researchers interested in high content analysis. The advantages and optimal utility of these formats will be covered with specific examples provided for each.2.Scratch assays were first used as models of wound healing for epithelial or mesenchymal cells . In thiset al. designed a 384-well format scratch assay using a 96-head pin tool array to scratch cell monolayers [A majority of researchers employing the scratch assay method utilize multiwell plates that contain 96 or fewer wells. Yarrow nolayers . They idnolayers .et al. [Simpson et al.  screenedet al. ,27.et al. describes the inherent limitations of scratch assays as an inability to achieve reproducible and quantitative results [in vitro results distinguished different levels of aggressiveness and invasiveness for three different cell lines which corresponded to their behavior in vivo [Kam  results . To addr results . The in  in vivo .i.e., to close the wound; (3) the assay surface can be coated with an ECM of choice prior to the experiment; and (4) the movement and morphology of the cells can be visually observed in real time and images captured throughout the experiment thereby permitting velocity measurements [Scratch assays provide several distinct advantages in that: (1) the assay can be performed in any readily available plate configuration; (2) cells move in a defined direction, urements . Howeverurements  and the urements -30. Moreurements ,29 and turements  and elecurements  exist asurements . While Eurements . Additiourements .3.Boyden originally described an assay technique as depicted in 50 values 20-fold less than that needed to abolish cell proliferation [Ogasawra and co-workers  utilizedferation . Howeveret al. [In an effort to improve sensitivity and throughput of transmembrane assays, Mastyugin et al.  used a met al. . While tet al. .et al. [An alternative assay that uses a measurement of transepithelial electrical resistance (TEER) to indicate the integrity and permeability of cell monolayers was described by Mandic et al. . In thiset al. .Transmembrane assays offer the distinct advantages of being able to analyze migration in response to a chemotactic gradient and can be used with adherent as well as non-adherent cells. Filter membranes may be coated with ECM proteins in an effort to better approximate physiological conditions. However, there are several drawbacks of these assays in that they are technically challenging to set up, the gradient is non-linear and equilibrates between both compartments over time , it is d4.Microfluidic systems such as microarrays, gradient devices, valved arrays and individually addressable channel arrays have recently emerged with the potential to be physiologically relevant and improve assay content to provide cell microenvironments for drug discovery . An examA microfluidic assay was used to demonstrate tumor cell migration through 3-dimensional matrices . In thisMicrofluidic chamber based assays can offer distinct advantages in situations where reagent availability is limited. A 800 nL volume of matrix is required to fill channels and only 1,000–2,000 cells are dispensed per channel which represents a 10–100 fold reduction from other methods without sacrificing assay robustness and better enabling the use of rare primary cells . However5.et al. explain: “The scratch process destroys the removed cells, which release their intracellular content into the medium; this process is also quite traumatic for the cells on the newly formed border. Indeed these border cells may become partially permeable as a result of the brutal tearing off of the adhesive junctions they maintain with their neighbors” [Cell exclusion zone assays originated from the need to study cell migration on an uncompromised surface uncoupled from contributions of cell damage and permeabilization that can arise from scratch wounds . Poujadeighbors” . Thus, tighbors” .Cell exclusion zone assays, as illustrated in The Oris™ Cell Migration Assay was validated for HTS using 3-day plate uniformity and replication of potency protocols established by Eli Lilly and the NIH Chemical Genomics Center which tested three assay plates with interleaved high, mid, and low inhibitor concentrations per day for 3 days . Z′ fact50 values for latrunculin A of 135 nM and 132 nM, respectively on MDA-MB-231 cells [To facilitate the use of automated liquid handling equipment for all steps of the assay, the Oris™ Pro Cell Migration Assays use a self-dissolving biocompatible gel (BCG) spotted in the centers of either 96- or 384-well plates instead of the silicone cell seeding stoppers. The BCG spots act as temporary barriers to prevent cells from settling and attaching during the seeding process and, once dissolved, reveal uniform areas into which cells may migrate . Vogt ha31 cells . Gough r31 cells .The Oris™ Cell Invasion Assays incorporate an ECM overlay in order to form a 3-dimensional environment to study invasion into the cell exclusion zone along the x, y, and z-axes . This asin vivo. Cell exclusion zone assays offer robust and reproducible data since the starting dimensions of the detection zone are accurately and precisely positioned in the assay wells. However, these assays are limited for use with adherent cells and cannot be used to establish chemotactic gradients.Cell exclusion zone assays offer the distinct advantage of not damaging the cells or the ECM as occurs in scratch assays. This assay format also allows continuous visual assessment of the cells throughout the experiment with the ability to acquire multiplexed data unlike the transmembrane assays where the filter restricts observation. In this way, information can be collected regarding morphology, velocity, distance and direction of migrating or invading cells as well as additional phenotypic effects of test compounds. In the Oris™ Cell Invasion Assays, the cell monolayer is entirely surrounded by the ECM thus reflecting a more physiologically relevant environment in which to study cell invasion and the effects of potential cancer therapeutics. In contrast, Brekhman and Neufeld  point ou6.et al. [et al. [et al. [Several recent studies examined the effects of oncogene expression or signal transduction mechanisms on cell motility in parallel scratch and transmembrane assays and found a good correlation of results. Valster et al.  demonstr [et al.  found th [et al. . RNA int [et al. . Bauer e [et al.  examined [et al. .et al. [et al. [Studies also compared cell exclusion zone assays to either scratch and/or transmembrane assays. Jiang et al.  demonstr [et al.  reported [et al. .Researchers at Platypus Technologies compared MDA-MB-231 cell migration on collagen I coated surfaces using both the Oris™ Cell Migration Assay and the scratch assay. Experiments were performed in parallel on four different days to compare the performance of each assay. For each independent experiment, the average area closure achieved using the Oris™ Cell Migration Assay  ranged fet al. [in vitro transmembrane invasion assays favor cells with spindle cell morphology and might be less appropriate for cells with epithelioid morphology. They mention that epithelioid cells might be more dependent on mechanisms of motility that rely upon influences from 3-dimensional microenvironments [There are also examples in which alternate assay formats for studying cell migration or invasion can give different, yet complementary results. Attoub and co-workers  used scret al.  examinedet al. . Howeveret al. . Cells iet al.  which pret al.  point ouronments . Table 17.in vitro and their efficacy as cancer therapies in human clinical trials [There has been a disappointing lack of correlation between compounds that inhibit migration and invasion l trials ,54. Howel trials . Carraghl trials  discussein vivoOffer ECMs and 3-dimensional environments to mimic cell behavior Allow for real-time visualization of cellsi.e., high-content analysis)Permit phenotypic and multiparametric analysis of cells (Facilitate high-throughput screening (automated liquid handling and high content imaging)To accelerate discovery of therapeutics affecting cell motility, cell-based assays should be utilized that:When researchers utilize only single target HTS assays, they obtain limited information on how potential therapeutics may influence complex multifaceted events such as tumor cell migration and invasion. As drug discovery screening continues to transition from biochemical to cell-based assays and from high throughput to high content screening, 3-dimensional culture technologies and phenotypic screens will become essential for increasing the relevance of screening assays ,19. Prof


Dear Editor,We read with great interest the article by Hossain et al., in April 2013 “Is discontinuation of clopidogrel necessary for intracapsular hip fracture surgery? Analysis of 102 hemiarthroplasties”. An increasing number of patients presenting for anesthesia are taking clopidogrel and an even greater number are on combination antiplatelet therapies, potentially increasing the risk of intraoperative and perioperative bleeding.Hossain et al.  found no® Platelet Mapping™ (TEG-PM) assay for preoperative assessment of platelet ADP receptor inhibition. Collyer et al. [It is our opinion that correlation between platelet function and responsiveness to clopidogrel is of paramount importance. There is significant individuality in patient responsiveness to clopidogrel, suggesting that an individualized, evidence-based approach is needed to assess the risk of adverse outcomes in patients receiving regular clopidogrel therapy. We suggest the use of the Thrombelastograph r et al. , 3, provWe assessed the ability of TEG-PM to detect preoperative platelet function secondary to clopidogrel and/or aspirin therapy . In an eTwo important findings were: (1) a trend towards platelet recovery by day 5 off clopidogrel, (2) 60 % of the patient population were not effectively inhibited while on therapy . In a pr


The high prevalence of cardiovascular disease (CVD) is largely attributable to the contemporary lifestyle that is often sedentary and includes a diet high in saturated fats and sugars and low ingestion of polyunsaturated fatty acids (PUFAs), fruit, vegetables, and fiber. Experimental data from both animals and humans suggest an association between increased dietary fiber (DF) intakes and improved plasma lipid profiles, including reduced low density lipoprotein cholesterol (LDL-C) concentrations. These observations underline that the intake of DF may protect against heart disease and stroke. Dietary fibers (DF) are highly complex substances described as nondigestible carbohydrates and lignins resistant to digestion and absorption in the small intestine . CommonlFruit, vegetables, wholegrains, and cereals are the major sources of DF components. Total DF can be divided into two groups: viscous fiber  and non-viscous fiber .DF are indigestible substances resistant to human digestive enzymes without nutritional or energetic value . DF was an almost unknown phrase and fibers were considered only annoying intestinal wastes until the 1970s when a wide range of potential therapeutic applications were suggested .The high prevalence of cardiovascular disease (CVD) is largely attributable to the contemporary sedentary lifestyle combined with a diet high in saturated fats and sugars, and low ingestion of polyunsaturated fatty acids (PUFAs), fruit, vegetables, and fiber. Epidemiological studies have confirmed a strong association between fat intake, especially saturated- and transfatty acids, plasma cholesterol levels, and rate of coronary heart disease (CHD) mortality . In contThe term DF includes a wide range of molecular structures, a highly complex mixture of different non-starch polysaccharides (NSPs), which include cellulose, β-glucans, hemicellulose, pectins, gums, polysaccharides of algae (agar and carrageenan) and lignin  ,10. Sourd-galacturonic acid is a principal constituent. They are structural component of plant cell walls and also act as intercellular cementing substances. The backbone structure of pectin is an unbranched chain of axial-axialα-(1-4)-linked d-galacturonic acid units. Pectin is highly water soluble and is almost completely metabolized by colonic bacteria. Other NCPs include gums, mucillages, and algal polysaccharides. Lignin is a cross-linked racemic macromolecule, a three-dimensional aromatic polymer of phenylpropane derivatives (containing about 40 oxygenated phenylpropane unit), and is associated with cellulose in plant cell walls. Lignins vary in molecular weight and methoxyl content. Due to strong intramolecular bonding which includes carbon to carbon linkages, lignin is very inert [The two major classes of DF are polysaccharides and lignin. Polymers of phenylpropane, are found in most plant structures in association with cellulose, and is the most widespread organic molecule on Earth and the major component of plant cell walls. It is a linear polymer made up of 10,000 to 15,000 glucose molecules bonded in a 1→4 glycosidic linkage. Cellulose molecules contain many polar hydroxyl groups, which allow them to interact with adjacent molecules to form fibers. These fibers are structurally strong and resistant to chemical attack. The more important non-cellulosic polysaccharide (NCPs) is hemicellulose and pectic substance. Hemicellulose are cell wall polysaccharides solubilized by aqueous alkali, which contain backbones of β-1,4-linked pyranoside sugar, and differ from cellulose in size (less than 200 sugar residues). The hemicelluloses are subclassified on the basis of the principal monomeric sugar residue . Pectin ry inert .The soluble DF has strong probiotic characteristics, which can be considered a non-digestible food ingredient that can selectively stimulate the growth and metabolic activity of a limited number of microbial groups, important for the proper functioning of the body. Recently, there has been a further substantial revision of the view of what exactly constitutes DF with the emerging recognition of the contribution of resistant starch and oligosaccharides to ‘fiber’ action . It was Consumption of DF has been associated to lower risk of CVD for some time , but theSoluble fiber clearly lowers cholesterol to a small but significant degree and one would expect that this would reduce coronary heart disease (CHD) events . The spevs. those who eat them rarely [Many epidemiological studies examined the relationship between wholegrain consumption and CHD. In these studies the researchers concluded that a relationship between wholegrain intake and CHD is seen with at least a 20% and perhaps a 40% reduction in risk for those who eat wholegrain food habitually m rarely . Notwithm rarely .et al. [A study about the relationship of long-term intake of dietary fiber by 68,782 women showed a substantial lowering of relative risk (0.53) for women in the highest quintile of fiber consumption (22.9 g/day) compared with the lowest (11.5 g/day). These intakes are low compared to those recommended by health authorities 30 g/day). Only the effect of cereal fiber was significant. However, the question of the relationship of other contributors to the effects of DF  and CHD risk remains unanswered. Elevated plasma total and low-density lipoprotein cholesterol (LDL-C) concentrations are established risk factors for coronary morbidity and mortality. There are abundant human and animal data showing that diets high in soluble fiber lower plasma cholesterol . One pop g/day. Oet al.  observedet al. ,34. Any et al. . It was et al. . This woet al. ,37.An analysis of dietary factors and cardiovascular risk performed in a sample of 3452 Swiss adults demonstrate that a healthy diet characterized by high consumption of dietary fiber was associated with lower rates of serum triglycerides and higher values of high density lipoprotein cholesterol (HDL-C) . An imprThe American Heart Association (AHA) recommends a total dietary fiber intake of 25 g/day to 30 g/day from foods (not supplements) to ensure nutrient adequacy and maximize the cholesterol-lowering impact of a fat-modified diet . Finallyet al. [et al. [et al. [vs. 42.4 g/day) was 0.82 . The authors acknowledge that a greater intake of wholegrains was related to an overall healthier diet and lifestyle. They performed stratified analyses to address potential confounding by healthy participant characteristics and found no discrepant results with respect to the main finding of protection from heart disease with wholegrains in subgroups including never smokers and non-drinkers.Wholegrain consumption has been shown to be linked to improvements in body mass index (BMI) ,43 insulet al.  did not et al. –52. The  [et al.  carried  [et al. –56 which [et al.  followedvs. those who eat them rarely [A further review has also reported the results of more recent prospective studies of wholegrain consumption The reviewers concluded that a relationship between wholegrain intake and CHD is seen with at least a 20% and perhaps a 40% reduction in risk for those who eat wholegrain food habitually m rarely .Jacobs and Gallaher consider that, given the wide variability in study designs, an estimated risk reduction of 20–40% is ‘impressively robust’. Study participants included men and women; participants from the US and Norway. Findings, which were consistently positive for wholegrains, occurred using a variety of different data collection methodologies.et al. [In conclusion, Jacobs and Gallaher and other reviewers find that there is good evidence that wholegrain foods substantially reduce the risk of CHD. Whether all grains are equal in this respect cannot be concluded from these studies, nor can the effectiveness of different parts of the grain. Results from Jensen et al.  suggest et al. [vs the lowest quintile). However, the inverse relation remained  suggesting that other constituents may confer additional protection. The authors acknowledge that their findings may not be generalizable to other populations (98% of their cohort were white).Liu et al.  examinedet al. [vs the lowest quintiles). Mozaffarian et al. [Steffen et al.  did incln et al.  followedet al. [Fung et al.  examinedIn conclusion, there are few studies to date that specifically examine the relationship between wholegrain intakes and risk of stroke. The studies described above have shown mixed results when adjustment has been made for potential confounders. However, the trends are strongly suggestive of a protective effect of wholegrain on risk of stroke.et al. [A number of researchers have found that a higher intake of cereal fiber is associated with a lower risk of CHD, although results tend not to be statistically significant after adjusting for multiple confounding factors ,53,60–62et al.  pooled tet al. [et al. [i.e., wholegrain intake) . Other cereal fibers had a non significant influence , other cold cereals , cooked cereals ). The authors did not state whether these other cold or cooked cereal fibers were wholegrain or refined grain products.Anderson et al.  also per [et al. . After aet al. [In contrast, Jensen et al.  investiget al. [Bazzano et al.  examinedet al. [vs the lowest quintile of total dietary fiber intake was observed for a lower BMI in those with the highest intake. For women, only vegetable fiber displayed an association. To date, detailed clinical data and the incidence of cardiovascular events in relation to fiber intake from this study have not been published.Lairon et al.  investiget al. [et al. [et al. [et al. [et al.’s cohort, Ness and co-workers acknowledge that dietary advice stopped after 2 years and recent data were collected when around half of the original participants had died. The researchers cannot discount the possibility that the diets of those who survived are different from those who did not. Follow-up diet was assessed with a limited number of questions focused on fiber intake; questionnaire data were not collected on other aspects of current diet and it is thus possible that important differences in diet were not detected. The authors concluded that the failure of longer term data to confirm the initial but non-significant increased risk of CHD deaths in men who were given advice to eat more cereal fiber, suggested that increased fiber intake was not harmful. The results do, however, suggest that increasing cereal fiber intake by a small degree does not confer any immediate survival advantage. It must be borne in mind, however, that this was a secondary prevention trial of CHD, in which the influence of environmental factors on the pathological process is likely to be weaker than in a primary prevention trial [Ness et al.  followed [et al. . Between [et al.  allocate [et al.  followedon trial .et al. [Jacobs et al.  argued tThese results highlight the emerging ‘wholegrain story’ which argues that health benefits stem from more than just the fiber; the wholegrain is nutritionally more important because it delivers a whole package of nutrients and phyto-protective substances that may work synergistically to promote health . Thus alThere is little epidemiological data on the association between soluble fiber and CHD even though soluble fiber clearly lowers serum cholesterol concentration and also lowers glucose in diabetics. A cholesterol-lowering claim is allowed for oat-derived beta glucan  by the United States Food and Drug Administration .In a study of female health professionals, fiber was associated with protection from heart disease . The asset al. [In a secondary prevention study in men with angina, Burr et al.  found noet al. [A meta-analysis of eleven clinical intervention trials  involvinet al.  confirmeFurthermore, two other well-known cardiovascular risk factors (type 2 diabetes and obesity) seem to be influenced by intake of DF. Vegetarians who consume a high-fiber lacto-ovo vegetarian diet appear to have a lower risk of mortality from diabetes-related causes compared to nonvegetarians . ConsumpA number of recent studies give novel insights that might help establish a metabolic link between insoluble DF consumption and reduced diabetes risk. Potential candidates are improved insulin sensitivity and the modulation of inflammatory markers, as well as direct and indirect influences on the gut microbiota . A breakCurrently, the American Diabetes Association recommends that diabetic patients consume 14 g/1000 kcal/day of fiber because a high amount of fiber is necessary to improve glycemic control. This amount is between 2 and 3 times higher than that consumed by individuals in many developed countries .The association between fiber intake and obesity or CHD is confirmed by several epidemiological studies. DF can modulate body weight by various mechanisms. Fiber-rich foods usually have lower energy content, which contributes to a decrease into the energy density of the diet . Foods ret al. [2; one moderately obese group, with a BMI between 27.1 and 40 g/m2; and one severely obese group, with a BMI > 40.0 kg/m2). These authors showed that fiber intake was significantly higher in the normal weight group and was inversely associated with BMI after adjusting for several potential confounders . The CARDIA (Coronary Artery Risk Development in Young Adults) study, a multicenter population-based cohort study carried out over 10 years, examined 2909 young individuals to determine the relationship between total DF intake and plasma insulin concentrations, weight and other CVD risk factors. After adjusting for BMI and multiple dietary  and potential non-dietary confounders , the study reported an inverse association between total fiber intake, plasma insulin concentrations and body weight gain [Alfieri et al.  assessedght gain  suggestiResults from all epidemiologic studies described above underline that the intake of wholegrain foods clearly protects against CHD and stroke but the exact mechanism is not yet clear. Moreover, the intake of high carbohydrates (from both grain and non-grain sources) in large amounts is associated with an increased risk of CHD in overweight and obese women even when fiber intake is high, but this requires further confirmation in normal-weight women.