In 2015, birth testing for certain asymptomatic HIV-exposed infants was included in the South African National Consolidated Guidelines for the first time.in utero and intrapartum HIV-infected infants failed to recognise:The notion that a single HIV polymerase chain reaction (PCR) test performed at 6 weeks of age would detect virtually all 4the HIV-related mortality that occurs prior to testing at 6 weeks of agethe reduced sensitivity of HIV PCR tests as a consequence of the increase in the number and duration of drugs used for prevention of mother-to-child transmission of HIV (PMTCT) prophylaxis.There is increasing evidence that both fixed dose combination (FDC) maternal PMTCT prophylaxis and daily dose nevirapine (NVP) infant prophylaxis (Option B or B+) contribute towards reduced detection of perinatal HIV infection at 6 weeks of age. The literature demonstrates that:in utero infected infants at 5 days and 2 weeks of age respectively.6 No HIV PCR sensitivity data for 6-week-old HIV-exposed infants, tested at discontinuation of 6 weeks of daily dose NVP, are available.a single perinatal dose of NVP reduced viral load to below the limit of detection in 38% and 17% of 7the probability of a positive HIV PCR at age 6 weeks in perinatally HIV-infected infants is decreased with multi-drug maternal and/or infant PMTCT prophylaxis8in non-breastfed infants, HIV DNA and RNA PCR sensitivity at 1 month of age for perinatally infected infants was 89%9in formula-fed infants who received 6 weeks of postpartum zidovudine (AZT), with or without other antiretrovirals, 32% of intrapartum-infected infants tested HIV DNA PCR negative at 6 weeks of age but tested positive at 3 months of age10prophylaxis reduces HIV DNA concentrations at birth complicating early identification of infected infants for initiation of early treatment12there are case studies illustrating the challenges of ‘false negative’ and ‘indeterminate’ HIV PCR results in early infant diagnosis in the context of current PMTCT prophylaxis and calling for revised diagnostic guidelines.in utero and intrapartum HIV infections in the context of current PMTCT interventions. It is too late to diagnose infants who die prior to 6 weeks of age and to achieve early combination antiretroviral therapy (cART) initiation by 7.4 weeks of age as was done on the Children with HIV Early Antiretroviral Therapy (CHER) trial to reduce early morbidity and mortality.13 It is too early to diagnose in utero and intrapartum infections suppressed by daily dose NVP and/or maternal prophylaxis via the placenta and/or breastmilk.The 6-week test, conveniently scheduled at the same time as the 6-week expanded programme for immunisation (EPI) visit, is therefore too late and too early to detect all in utero and intrapartum HIV infections) can never approach 100% because it detects in utero infection only and cannot detect intrapartum infection, namely an infection transmitted at the time of labour and delivery and essentially in the window period at birth. Prior to the implementation of PMTCT and the use of standardised HIV assays, Dunn et al.’s meta-analysis demonstrated that 38% of all perinatal infections were detectable at birth.14 With World Health Organization (WHO) Option A prophylaxis and a single dose of NVP at birth for infants, Lilian et al. demonstrated that 76% of all early HIV infections were detectable at birth.15 This increase was attributed to more sensitive viral detection assays and a proportional increase in in utero to intrapartum infections as a result of PMTCT prophylaxis targeting intrapartum infections during late pregnancy and delivery. As the majority of women deliver in health facilities in South Africa, identifying all HIV-infected women at delivery coupled with birth HIV PCR testing would yield three-quarters of all perinatal HIV infections with close to 90% coverage.The HIV PCR sensitivity at birth for detecting perinatal HIV infections , low birth weight (LBW < 2500 g) or symptomatic HIV-exposed neonates and those born to women who were unbooked or received a late diagnosis of HIV (e.g. at delivery) or received < 4 weeks of antiretroviral PMTCT prophylaxis or had viral loads > 1000 copies per millilitre or were co-infected with tuberculosis during pregnancy. The Western Cape PMTCT guidelines also recommend targeted birth testing that include additional and slightly modified high-risk factors.17 These high-risk factors predict vertical transmission but not necessarily in utero transmission, and therefore the birth HIV PCR test may be negative.The 2014 South African National Consolidated Guidelines recommend targeted birth testing, namely HIV PCR testing of only those HIV-exposed neonates identified as being at high risk of HIV transmission.in utero and intrapartum infections, on every HIV-exposed infant can be offset by following 2010 WHO guidelines to use HIV rapid tests (HRT) from 9 months of age.18 As the majority of HIV-exposed, uninfected infants demonstrate seroreversion by 9 months of age by testing HRT negative, only those with positive HRT would require HIV PCR tests.19 This approach reduces the number of HIV PCR tests required for symptomatic infants or those requiring testing post-cessation of breastfeeding.Universal birth testing of all HIV-exposed infants may be simpler to implement than targeted birth testing. The cost of performing two early HIV PCR tests, to detect To avoid morbidity and mortality, all positive HIV PCR results require urgent action to (1) confirm the HIV-infected status on a second blood sample and (2) initiate cART. Healthcare facilities require good communication with the laboratory to access positive results within 2–7 days and systems for patient follow-up to see all HIV PCR-positive patients as soon as possible. As neonates usually return to their primary healthcare clinic and not the maternity unit for follow-up after birth, ensuring that birth HIV PCR test results reach patients will be pivotal to successful implementation of birth testing. Point of care (POC) HIV diagnosis would facilitate same-day identification of HIV-infected neonates.The results of the second, confirmatory viral detection assay should not delay cART initiation but should be obtained as early as possible because, once cART is initiated, it becomes progressively more difficult to detect HIV either by HIV PCR or viral load testing. The availability of two different POC assays for detection of HIV would facilitate same-day identification and confirmation of HIV infection in neonates.1 fewer cases of intrapartum infection may be identified owing to the viral load lowering effect of the daily dose NVP infant prophylaxis. The 2014 National Consolidated Guidelines cater for this in high-risk infants only by recommending a third HIV PCR at 16 weeks of age where prolonged infant prophylaxis (e.g. daily dose NVP for 12 weeks) has been used.1 The probability that the sensitivity of diagnostic virological assays is affected by antiretroviral prophylaxis has prompted American guidelines to recommend an additional HIV PCR test be performed 2–4 weeks after combination antiretroviral infant prophylaxis has been discontinued if negative HIV PCR results were obtained during prophylaxis.20 As for all HIV-exposed and uninfected infants, HIV PCR testing is recommended whenever clinical features suggestive of HIV infection are present and 6 weeks after cessation of breastfeeding (if < 18 months old). If, at 6 weeks after weaning, the child is ≥ 18 months old, a HRT or HIV enzyme-linked immunosorbent assay (ELISA) test should be performed instead of an HIV PCR test.If the birth HIV PCR test is negative, an HIV PCR test at 10 weeks of age is recommended to detect as many cases of intrapartum infection as possible. If the HIV PCR test is repeated at 6 weeks of age, as per national guidelines,In an evolving PMTCT environment, ongoing monitoring is necessary to assess the impact of early diagnosis of HIV infection in neonates and to ensure that an evidence-based, effective diagnostic algorithm is deployed.


Escherichia coli O103 strains shed in the feces of cattle. We report here the genome sequences of 43 strains of enterohemorrhagic E. coli (EHEC) O103:H2 isolated from feedlot cattle feces. The genomic analysis will provide information on the genetic diversity and virulence potential of bovine EHEC O103.The enterohemorrhagic pathotype represents a minor proportion of the  Escherichia coli (EHEC). The organism is shed in the feces of cattle, which can contaminate the hide and then carcass at slaughter, creating the potential for foodborne illness in humans  and intimin (eae) genes, the combination of these and presence of other major virulence genes, including enterohemolysin (ehxA), vary among EHEC O103 strains isolated from bovine feces , genome size, and number of contigs per genome are summarized in We report here the draft whole-genome sequences of 43 strains of EHEC O103:H2 isolated from feces of feedlot cattle in the United States . BacteriThe 43 strains deposited here will add to the limited publicly available genetic information on EHEC O103. These genomes will be a valuable resource for investigations into the genetic features of this major foodborne pathogen, including research into the genome evolution of EHEC O103 compared to other EHEC serogroups, and insights into specific factors allowing for adaptation to a bovine host. This work will also provide an opportunity to study the potential virulence risk of these EHEC O103 strains to humans.The whole-genome shotgun sequence has been deposited at DDBJ/ENA/GenBank under the accession numbers listed in


Androgen deprivation therapy (ADT) has been the standard of care for advanced prostate cancer (PCa) for decades, yet men with this condition inexorably develop resistance to ADT and progress to metastatic castration-resistant PCa (mCRPC) . Second It was shown previously that AR transcriptionally upregulates multiple DNA damage response (DDR) genes -5, and tin vitro [in vitro. We used multiple subcutaneous and orthotopic in vivo CRPC models, including an ENZ-responsive PDX model to validate the superiority of lead-in combination ENZ+OLA treatment compared to concomitant administration [We initially identified a subset of 10 HR genes which were upregulated in CRPC in 2 independent public data sets, and showed that 7 of these upregulated genes were contained within a 15 HR gene subset which was downregulated by ENZ in microarray analysis [The causative mechanisms for susceptibility of cancers with BRCA mutations to PARP inhibitor — mediated synthetic lethality are complex, and recent studies have pointed to the need to expand the concept of BRCAness in tumors beyond germline BRCA1 or BRCA2 mutations to include mutations in genes that modulate HR and confer PARP sensitivity . Our rec


Persons who experience symptoms of psychosis following a brain injury live with a complex dual diagnosis that is often accompanied by substantial distress and disability. However, a comprehensive examination of the clinical presentation of PFTBI using standardised clinical measures has not been reported in the literature. This information is vital for accurate diagnosis and effective treatment.Patients with PFTBI (n =10) and schizophrenia (n =98) participated in a comprehensive clinical assessment that included the Structured Clinical Interview for DSM-IV-TR Axis I Disorders (SCID-I/P), the Positive and Negative Syndrome Scale (PANSS), the Scale for the Assessment of Positive Symptoms (SAPS), the Scale for the Assessment of Negative Symptoms (SANS), and the Thought Language and Communication Index (TLC).The clinical profiles of the PFTBI group met symptom/course criteria for: schizophrenia (n = 6), schizoaffective (n = 2), schizophreniform (n = 1), and paranoid psychosis (n = 1). PFTBI patients had a significantly reduced PANSS negative score relative to schizophrenia patients. No other significant differences between PFTBI and schizophrenia clinical profiles were found.The clinical profile of PFTBI appears to be comparable to schizophrenia/ schizoaffective disorder except with respect to negative psychotic symptoms. Reduced negative symptoms in PFTBI have previously been reported in a small number of case studies, and thus warrant further investigation as a diagnostic distinction in this patient group.


The carotid surgical experience of Cheltenham General Hospital over a 13 year period (1968-81) is presented. This includes 42 operations for stenosis, and 12 further operations for carotid body tumour, carotid aneurysm, subclavian steal syndrome and trauma to the internal carotid artery. The operative techniques and complications are briefly discussed and reasons advanced for a more agressive approach to the problems of extra-cerebral carotid disease in this country.


The early differentiation of Alzheimer’s disease (AD) from frontotemporal dementia (FTD) may be difficult. The Tower of London (ToL), thought to assess executive functions such as planning and visuo-spatial working memory, could help in this purpose.Twentytwo Dementia Centers consecutively recruited patients with early FTD or AD. ToL performances of these groups were analyzed using both the conventional statistical approaches and the Artificial Neural Networks (ANNs) modelling.p < 0.05) The use of hidden information contained in the different items of ToL and the non linear processing of the data through ANNs allows a high discrimination between FTD and AD in individual patients. However, the discriminant validity of AS checked by ROC curve analysis, yielded no significant results in terms of sensitivity and specificity (AUC 0.63). The performances of the 12 Success Subscores (SS) together with age, gender and schooling years were entered into advanced ANNs developed by Semeion Institute. The best ANNs were selected and submitted to ROC curves. The nonlinear model was able to discriminate FTD from AD with an average AUC for 7 independent trials of 0.82.Ninety-four non aphasic FTD and 160 AD patients were recruited. ToL Accuracy Score (AS) significantly (The use of hidden information contained in the different items of ToL and the non linear processing of the data through ANNs allows a high discrimination between FTD and AD in individual patients.


It  Specifications TableValue of the data•ilr transformation for other food compositions and interested researchers.The data include the sequential binary partition of fatty acid profiles in CoDa and could be useful for calculating balances and the •The correlation among fatty acids expressed in percentages may help other researchers for finding spurious relationships.•The information may facilitate the comparison of conventional multivariate techniques and compositional, regardless of the field, and promote international collaborations in data analysis.•Presentation in the Simplex can be an appropriated way of graphing compositional data and treatments’ effects.1ilr transformations based on it  Fatty acid profiles were obtained through analysis of their FAMEs by GC according to the procedures recommended in the Commission Regulation (EU) No 2015/1833 ilr transformed values (or coordinates)  were subjected to similar multivariate Cluster Analysis (based on the Euclidean distance and the Ward method) (The data matrix consisted of 10 rows (five treatments in duplicate) and 19 columns (fatty acids). Values were first tested for outliers and normality. The data were plotted in the Simplex , analysedinates)  [10], [1 method)  [12].TabCoDaPack v. 2.01.14 , XLSTAT 2014  were used for data processing and graph drawing.


Objective: To characterize various dysgraphic symptoms in parietal agraphia. Method: We examined the writing impairments of four dysgraphia patients from parietal lobe lesions using a special writing test with 100 character kanji (Japanese morphograms) and their kana (Japanese phonetic writing) transcriptions, and related the test performance to a lesion site.Results: Patients 1 and 2 had postcentral gyrus lesions and showed character distortion and tactile agnosia, with patient 1 also having limb apraxia. Patients 3 and 4 had superior parietal lobule lesions and features characteristic of apraxic agraphia (grapheme deformity and a writing stroke sequence disorder) and character imagery deficits . Agraphia with impaired character recall and abnormal grapheme formation were more pronounced in patient 4, in whom the lesion extended to the inferior parietal, superior occipital and precuneus gyri.Conclusion: The present findings and a review of the literature suggest that: (i) a postcentral gyrus lesion can yield graphemic distortion (somesthetic dysgraphia), (ii) abnormal grapheme formation and impaired character recall are associated with lesions surrounding the intraparietal sulcus, the symptom being more severe with the involvement of the inferior parietal, superior occipital and precuneus gyri, (iii) disordered writing stroke sequences are caused by a damaged anterior intraparietal area.


Intermittent catheterisation is the preferred method of managing the neurogenic bladder in patients with spinal cord injury. However, spinal cord physicians experienced problems when trying to implement an intermittent catheterisation regime in some spinal cord injury patients in the northwest of England. We present illustrative cases to describe practical difficulties encountered by patients while trying to adopt an intermittent catheterisation regime. Barriers to intermittent catheterisation are (1) caregivers or nurses are not available to carry out five or six catheterisations a day; (2) lack of time to perform intermittent catheterisations; (3) unavailability of suitable toilet facilities in public places, including restaurants and offices; (4) redundant prepuce in a male patient, which prevents ready access to urethral meatus; (5) urethral false passage; (6) urethral sphincter spasm requiring the use of flexible-tip catheters and α-drenoceptor–blocking drugs; (7) reluctance to perform intermittent catheterisation in patients >60 years by some health professionals; and (8) difficulty in accessing the urethral meatus for catheterisation while the patient is sitting up, especially in female patients. These cases demonstrate the urgent need for provision of trained caregivers who can perform intermittent catheterisation, and improvement in public facilities that are suitable for performing catheterisation in spinal cord injury patients. Further, vigilance should be exercised during each catheterisation in order to prevent complications, such as urethral trauma and consequent false passages. Health professionals should make additional efforts to implement intermittent catheterisation in female spinal cord injury patients and in those >60 years.


In the associated research article in vitro. We provide data demonstrating the use of nanoscale secondary ion mass spectroscopy (NanoSIMS) to identify specific ions in bulk dried NP. NanoSIMS is also used to visualize 40Ca microdomains in the z dimension of optic nerve that has been subjected to a partial optic nerve transection. The associated article 40Ca microdomains in optic nerve from animals treated with various nanoparticle preparations and provides further interpretation and discussion of the findings.Before using nanoparticles for therapeutic applications, it is necessary to comprehensively investigate nanoparticle effects, both  A sample raw NanoSIMS spectra of bulk dried NP and associated spatial resolution data demonstrate the ability to detect 56Fe within NP. NanoSIMS can be used to detect 40Ca microdomains in the z dimension of optic nerve following partial optic nerve transection.The oxidative stress indicator carboxymethyl lysine was assessed in an astrocyte-like immortalized Müller cell line (rMC1 cells). Cells were stressed with H2in vitro, assessed using ImageJ image analysis software  software .Fig. 1CeRaw NanoSIMS spectra  and specz-dimension of an optic nerve field of view (FOV) following partial optic nerve transection, collected using NanoSIMS. All procedures involving animals conformed to the National Health and Medical Research Council of Australia Guidelines on the Use of Animals in Research and were approved by the Animal Ethics Committee of The University of Western Australia   .Fig. 3Pa


Improving quality of maternal care as well as patients' safety are two important issues in health-care service. Therefore, this study aimed to assess the culture of patient safety at maternity units.This cross-sectional study was conducted among staffs working at maternity units in seven hospitals of Ilam city, Iran. The staffs included in this study were gynecologists and midwifes working in different positions including matron, supervisors, head of departments and staffs. Data were collected using the Hospital Survey on Patient Safety Culture (HSOPSC).This study indicated that 59.1% of participants reported fair level of overall perceptions of safety and 67.1% declared that no event was reported during the past 12 months. The most positively perceived dimension of safety culture was teamwork within departments in view of managers (79.41) and personnel (81.10). However, the least positively perceived dimensions of safety culture was staffing levels.The current study revealed areas of strength (teamwork within departments) and weakness  among managers and personnel. In addition, we found that staffs in Ilam's hospitals accept the patient safety culture in maternity units, but, still are far away from excellent culture of patient safety. Therefore, it is necessary to promote culture of patient's safety among professions working in the maternity units of Ilam's hospitals. Decreasing maternal mortality rate is recognized as an essential priority in health-care systems , 2. ImprThis cross-sectional study was conducted among staffs working at maternity units  in seven hospitals of Ilam University of Medical Sciences during year 2016. The staffs included in this study were gynecologists and midwifes working in several positions including managers, supervisors, head of departments and staffs. The total number of 350 staffs was invited to take part in the current study and 299 agreed to participate in the survey. Participants were informed about the aim, benefits and importance of the survey as well as their voluntary participation rights. Written informed consent was obtained prior to actual data collection. Data were collected using series of self-administrated questionnaires including socio-demographic characteristics and the Hospital Survey on Patient Safety Culture (HSOPSC). The socio-demographic and individual characteristics include age, gender, education, working hours per week, ward, organizational position, organizational tenure, work experiences in the current ward and type of employment. The Hospital Survey on Patient Safety Culture (HSOPSC) released by the Agency for Healthcare Research and Quality (AHRQ) to help hospitals assess the safety culture within their organization. The HSOPSC contains 42 items with 5 point Likert responses scale ranging from strongly disagree (1) to strongly agree (5). HSOPSC questionnaire contains both positively and negatively worded items. The average percent-positive score for each dimension was calculated and negative items were reverse scored prior to computing percent positive response. This questionnaire is designed to measure 12 dimensions of patient safety culture and these dimensions are: Supervisor/manager expectations and actions promoting safety (4 items), Organizational learning-continuous improvement (3 items), Teamwork within units (4 items), Communication openness (3 items), Feedback and communication about error (3 items), Non-punitive response to error (3 items), Staffing (4 items), Hospital management support for patient safety (3 items), Teamwork across hospital units (4 items), Hospital handoffs and transitions (4 items), Overall perceptions of safety (4 items), Frequency of event reporting (3 items) and Two outcome variables. The two outcome variables compose of “Overall perceptions of safety” and “Frequency of event reporting”. The HSOPSC questionnaire was validated in Persian language and shown a good validity and reliability to be used in Iran's hospitals . HoweverMean age of participants in this study was 33.52 (SD= 6.37) years old  and 84.1This study assessed culture of patient's safety in hospitals of Ilam University of Medical Sciences. Creating culture of patient safety requires assessment of safety culture using appropriate tool. This study measured culture of patient safety using HSOPSC questionnaire which is one of the best tools in this area . The curThe results of this study showed acceptable levels of overall perception of safety culture among both managers and personnel with higher perception among managers. This finding implies that health-care workers in Ilam's hospitals accept the patient safety culture in maternity units, but still are far away from excellent culture of patient safety. Maternal wards are among very important units in each hospital, which require special attentions. Therefore, it is necessary to promote culture of patient's safety among professions working in the maternity units of Ilam's hospital. There are several limitations in this study, which need to be addressed. One of the limitations in this study was lack of evidences in maternity units to compare the results. Therefore, future studies are recommended to assess the culture of patient safety in maternal care units using HSOPSC questionnaire. Another limitation in this study is use of self-reported data.International reports on maternal mortality rate has shown constant decreasing pattern during the last 15 years;Programs aimed at decreasing maternal mortality rate has always been a priority in the Iranian health system and some of these programs are promoting mother and child-friendly hospitals that offer maternal care using professional midwifery staffs;Promoting patients' safety and risk management are of great importance in maternity care units due to the fact that every procedure in this field will affect two individuals (both mother and baby).This study assessed culture of patient's safety in hospitals of Ilam University of Medical Sciences for the very first time and revealed areas of strength and weakness among managers and personnel;The most positively perceived dimension of safety culture was attributed to “teamwork within department” among both managers and personnel. While, the least positively perceived dimension was “staffing” and “non-punitive response error”, respectively;Positive perception toward “staffing” was higher among managers compared to personnel and “non-punitive response error” was almost equal between two groups.The authors declare no competing interests.


This article evaluated the autosomal dominant polycystic kidney disease (ADPKD) modifiers to chronic kidney disease (CKD) such as allelic and nonallelic agents. Also, our study evaluated the role of the endotelin-1 which considered in some studies as the predictor of the ADPKD.Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common life threatening inherited kidney disease which its prevalence is approximately 0.5 to 1 in 1000 subjects of general population ,3. FurthThe heterozygous mutations in PKD 1 (approximately 78%) and PKD 2 (approximately 13%) genes are responsible for the vast majority of the ADPKD incidence and no mutation detected (NMD) ADPKD exists in the other patients (9%)  and increase in kidney size are as modifiers. Furthermore some laboratory factors such overt hematuria, macroalbominuria, high level of serum copeptin concerned as predicting factor for chronic kidney disease (CKD) in ADPKD . The vasEndotelin-1 (ET-1) is another agent that has been reported to exert multiple effects on renal function . Some stIn summary, the progression of ADPKD to CKD depends on multiple factors include allelic and non-allelic factors that can be use in prevention of the CKD in these patients and need further researches for achieve to this issue.The authors contributed equally to the manuscript.The authors declared no competing interests.Ethical issues  have been completely observed by the authors.None.


Nature Communications 10.1038/s41467-017-00717-y, published online 18 September 2017Correction to: The original version of this Article omitted the Acknowledgements section:“This work was sponsored by the Australian Research Council (ARC) Laureate Fellowship (FL120100029) and the Centre of Excellence program (CUDOS CE110001010). We acknowledge the support of the ANFF ACT.”This has now been corrected in both the PDF and HTML versions of the Article.


Studies have shown that physiologic factors such as decreased LAA velocity and increased stasis are associated with thrombus formation in the LAA and may be better predictors for stroke. Currently available diagnostic tools such as TEE, however, are limited as they do not completely assess the complex 3D LAA blood flow and are invasive. As a result, the impact of AF on global and regional atrial 3D flow dynamics is poorly understood. In addition, it is unclear if the stimulus for LAA thrombus formation is driven by hemodynamic changes in the LAA alone or more broadly associated with AF induced flow alterations affecting the entire left atrium (LA). It was the aim of this study to test the potential of novel 4D flow MRI tools  for the characterization of LA and LAA flow dynamics.Atrial fibrillation (AF) is associated with increased risk of stroke due to development of left atrial appendage (LAA) thrombus. Clinical risk scores (CHA4D flow MRI was employed to measure in-vivo time-resolved 3D blood flow velocities in 75 subjects: 30 AF patients in sinus rhythm (AF-sinus), 30 AF patients in AF (AF-afib) and 15 normal controls. Data analysis included 3D segmentation of the LA and LAA to isolate the velocity data in the LA and LAA figure . Peak veLA and LAA mean velocity, stasis, and peak velocity were significantly different between controls, AF-afib patients, and AF-sinus patients  not just in the LAA but in the in the entire left atrium. An individual assessment of LAA and LA flow may thus help to better identify patients at risk for thromboembolism beyond current clinical risk scores.


Phaseolus vulgaris L.). In this study, 41 rhizobia isolates were obtained from the root nodules of MAC 13 and MAC 64 climbing beans grown in upper and lower midland agro-ecological zones of Eastern Kenya. Eastern Kenya was chosen because of its high production potential of diverse common bean cultivars. The rhizobia isolates were characterized phenotypically on the basis of colony morphology, growth and biochemical features. Rhizobia diversity from the different regions of Eastern Kenya was determined based on the amplified ribosomal DNA restriction analysis (ARDRA) of PCR amplified 16S rRNA genes using Msp I, EcoR I, and Hae III restriction enzymes. Notably, native rhizobia isolates were morphologically diverse and grouped into nine different morphotypes. Correspondingly, the analysis of molecular variance based on restriction digestion of 16S rRNA genes showed that the largest proportion of significant (p < 0.05) genetic variation was distributed within the rhizobia population (97.5%) than among rhizobia populations (1.5%) in the four agro-ecological zones. The high degree of morphological and genotypic diversity of rhizobia within Eastern Kenya shows that the region harbors novel rhizobia strains worth exploiting to obtain strains efficient in biological nitrogen fixation with P. vulgaris L. Genetic sequence analysis of the isolates and testing for their symbiotic properties should be carried out to ascertain their identity and functionality in diverse environments.The increasing interest in the use of rhizobia as biofertilizers in smallholder agricultural farming systems of the Sub-Saharan Africa has prompted the identification of a large number of tropical rhizobia strains and led to studies on their diversity. Inoculants containing diverse strains of rhizobia have been developed for use as biofertilizers to promote soil fertility and symbiotic nitrogen fixation in legumes. In spite of this success, there is paucity of data on rhizobia diversity and genetic variation associated with the newly released and improved mid-altitude climbing (MAC) bean lines ( Rhizobia bacteria play a significant role in provision of agricultural ecosystem services due to their ability to form symbiotic association with a wide range of leguminous plants that results in biological nitrogen fixation . Some ofMimosa genus  and chickpeas (Cicer arietinum L.), while Bradyrhizobium strains are often found to nodulate soybeans (Glycine max L. Merrill), cowpeas (Vigna unguiculata L.) and green grams (Vigna radiata L.). However, in some cases, cross inoculation by both genera has been documented in some legumes such as mungbeans (V. radiata L.) and this is considered advantageous and economical to farmers growing different types of legumes (Rhizobia-legume symbiosis is a host-specific association and hence the need to determine the strains and the diversity of rhizobia associated with specific type of legume for better exploitation of the benefits associated with the rhizobia biofertilizers . Rhizobisa genus . Until racterium . However legumes .P. vulgaris L.), the MAC beans establish a mutual and a beneficial symbiotic partnership with Rhizobium leading to the formation of root nodules that catalyze nitrogen fixation from the atmospheric air , restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) have been used to evaluate the diversity of rhizobia . In addiIn this study, we hypothesized that there exists a large heterogeneity among the rhizobia isolates nodulating MAC beans grown in different AEZ with contrasting environmental conditions in Eastern Kenya. The aim of this study was to isolate, characterize morphologically and genetically and determine the diversity of the rhizobia populations associated with MAC 13 and MAC 64 climbing beans grown in different AEZ of Eastern Kenya.Field trapping of rhizobia was carried out during the first rainy season between March and August 2015 in four different sites of Eastern Kenya with contrasting agro-climatic conditions using MAC 13 and MAC 64 climbing beans as the host plants. Eastern Kenya was chosen because of its high production potential of diverse common bean cultivars . The fouTen healthy and undamaged root nodules of MAC 13 and MAC 64 climbing beans from each site were selected and used in the isolation of rhizobia following the standard protocols described by Rhizobium. Based on the differences in the observed morpho-cultural, Gram stain and growth features, the isolates were placed into different morphotype groups.Using the standard microbiological techniques described by Hae III, EcoR I and Msp I . Three reference strains; Rhizobium tropici CIAT 899, Rhizobium etli USDA 2667 strain and Rhizobium leguminosarum strain 446 were used. The three reference Rhizobium strains were obtained from the Microbiological Resource Centre (MIRCEN), University of Nairobi, Kenya.The molecular study involved 41 rhizobia isolates and was carried out at the Department of Microbiology, Kenyatta University. The analysis of genetic relatedness of the native isolates from Eastern Kenya was carried out using amplified rDNA restriction analysis (ARDRA) of the 16s rRNA genes using three restriction enzymes The extraction of total genomic DNA of rhizobia isolates was carried out following modification of a method outlined by Escherichia coli , 0.6 μl dNTP (10 mM), 0.3 μl of each Y1 and Y3 primers (10 μM), 0.6 μl 5% Tween 20, 0.15 μl Taq DNA polymerase (Biolabs) and 1.5 μl of DNA template. The sequences of the primers used for PCR amplification of the 16S rRNA gene were: Y1 forward primer (5′-TGGCTCAGAACGAACGCTGGCGGC-3′) that corresponds to positions 20–43 and Y3 reverse primer (5′-TACCTTGTTACGACTTCACCCCAGTC-3′) corresponding to positions 1482–1507 for 16S rDNA sequence of hia coli . A negatHae III, EcoR I, and Msp I restriction enzymes (Biolabs) (Polymerase chain reaction products of the isolates and reference strains were digested using Biolabs) . A masteH) was also used to determine the rhizobia diversity in different AEZ of Eastern Kenya.The data on rhizobia genetic diversity based on the different band patterns formed after PCR restriction were coded in binary form and analyzed as described by Table 1). Morphotype III was the most abundant (29%) and morphotype II accounted for a partly 2% of the total number of isolates. All isolates were Gram negative rods and did not absorb Congo red dye when incubated in the dark on YEMA-CR medium . Notably, all isolates turned BTB indicator from deep green to yellow when grown on YEMA-BTB indicating that the isolates were acid producers and fast growers . The colony color varied with milky white, cream white, cream yellow and watery colonies being observed which were either opaque or translucent with either firm gummy or smooth mucoid texture . Most of the isolates had a mucoid texture due to the production of excess EPSs. As expected, all isolates had an entire colony margin but the colony elevation varied consistently with convex and raised colonies being observed on YEMA media . The morphological characteristics of the reference strains Rhizobium tropici CIAT 899 and Rhizobium etli USDA 2667 were closely similar to those of morphotype III while Rhizobium leguminosarum strain 446 closely resembled morphotype V (Table 1).From the field trapping experiment using MAC 13 and MAC 64 climbing beans, 41 pure rhizobia isolates were obtained, which were placed in to nine distinct morphotype groups based on their morpho-cultural and biochemical features . Restriction of the 16S rRNA amplicons of the climbing bean rhizobia isolates using enzymes EcoR I , Hae III  and Msp I  produced multiple band patterns. Restriction enzyme Msp I produced the most diverse polymorphic patterns.The PCR amplification of the 16S rDNA region produced a single band of approximately 1500 bp (Na = 1.92 ± 0.08) and effective alleles (Ne = 1.55 ± 0.10) compared to other populations (Table 2). Rhizobia isolates from ELM had the highest percentage polymorphic loci (92.31% P) while isolates from EUM and TLM recorded the lowest at 61.54% P. The mean Shannon–Wiener diversity (H) estimate showed that the four rhizobia populations from Eastern Kenya were genetically diverse with rhizobia population from ELM having the highest genetic diversity estimate of H = 0.47 (Table 2). The rhizobia population from zone TLM recorded the lowest genetic diversity estimate of H = 0.31. The average expected heterozygosity (He) varied among the four populations and ranged from 0.21 for rhizobia population from TLM to 0.32 for ELM population (Table 2).Based on ARDRA analysis of 16S rRNA amplicons, rhizobia population from ELM had the highest average number of different alleles  within populations . The variation among the rhizobia populations from the four AEZ (2.5%) and two Counties (0.5%) were, however, not significant (Table 3).Analysis of molecular variance (AMOVA) for the four rhizobia populations in Eastern Kenya showed a significantly high genetic variation . Isolates from TUM zone were the most distributed and appeared in all the four quadrants, while isolates from ELM zone were the least distributed .The principal coordinate analysis (PCA) of 41 native isolates from the four zones of Eastern Kenya and three rhizobia reference strains showed considerable differentiation . Rhizobia populations from ELM and TUM clustered together with a bootstrap value of 85% while rhizobia populations from EUM and TLM clustered with a bootstrap value of 82%.Based on the pairwise population matrix of Nei unbiased genetic distance and Euclidian similarity index, the neighbor joining dendrogram clustered rhizobia populations from Eastern Kenya into two main groups . Cluster I comprised of majority of the isolates while cluster III had only one isolate ELM5. The reference strain Rhizobium tropici CIAT 899 which was placed in cluster I, clustered together with native rhizobia isolates ELM33 and TLM32, indicating their close genetic relationship. Native isolates TUM26, TLM28, and EUM23 clustered together despite originating from different AEZ of Eastern Kenya. Rhizobium etli USDA 2667 strain was also placed in cluster I and was grouped together with TLM27 and TUM 41 isolates . The native isolate EUM7 clustered closely to Rhizobium leguminosarum strain 446 in cluster II but in a separate sub-branch.Based on the genetic distance after amplified rDNA restriction analyses and Euclidian similarity index, the phylogenetic tree clustered the native rhizobia isolates into three main clusters I, II, and III  estimate of rhizobia populations from Eastern Kenya based on genetic distance of ARDRA pattern analysis showed that the isolates were genetically diverse with rhizobia population from ELM zone having the highest genetic diversity estimate compared to TLM zone, which recorded the lowest diversity. This variation in rhizobia diversity could be due to the differences in agroclimatic and soil conditions of the four sites studied in Eastern Kenya. These results reinforce the promiscuous nature of P. vulgaris L. to nodulate with diverse strains of Rhizobium  genetic variation distributed within regions (89%) than among regions (11%) in S. meliloti and S. medicae obtained from drought and salt affected regions of Morocco. Moreover, The analyses of molecular variance (AMOVA) based on amplified 16S rDNA restriction profiles showed a highly significant genetic variation of native rhizobia isolates within rhizobia populations and not among the four populations nor across the two regions (Counties) of Eastern Kenya. Based on ARDRA fingerprints, the low level of genetic differentiation of MAC bean rhizobia could suggest that the rhizobia population within the region is weakly structured. This could be due to the absence of physical barriers to limit gene flow . Human aAgrobacterium, Bradyrhizobium, Rhizobium, and Ensifer). Based on the evolutionary relationship of 41 isolates inferred using Neighbor-Joining method, some of the native isolates clustered closely with the three reference rhizobia strains used in this study. The phylogenetic tree showed a diverse genetic variation among the native rhizobia isolates as evident by the different clusters and sub-clusters.The principal coordinate analysis (PCA) also showed low level of genetic differentiation of native rhizobia isolates from Eastern Kenya and thus showing congruent results with the dendrogram. However, the grouping and distribution patterns of the native isolates did not correspond to the AEZ, and this is an indicative of the likelihood of a common evolutionary origin of the isolates. Similarly, Rhizobium tropici CIAT 899 and R. etli USDA 2667 in cluster I. For instance, native isolates TLM 27 and TUM 41 clustered together with R. etli USDA 2667 while isolate TLM 32 clustered with R. tropici CIAT 899. These indicate the close genetic relationship between some of the native rhizobia isolates to R. tropici CIAT 899, R. etli USDA 2667 and R. leguminosarum strain 446  genetic variation distributed within populations (97%) than among populations (2.5%) in four AEZ of Eastern Kenya. Despite the high diversity shown within the rhizobia population, the Pairwise Population Matrix of Nei unbiased genetic distance of rhizobia populations in the study region showed a narrow range, which is an indication of a weakly structured genetic population and a highly conserved genetic structure of rhizobia within the study area. Further molecular studies using either full or partial gene sequences of bacterial genome, which is more sensitive and has a higher resolution power, needs to be carried out to establish the true diversity of native rhizobia isolates up to the species and strain levels.In this study, 41 rhizobia isolates were obtained from the root nodules of mid-altitude climbing (MAC) beans and based on morpho-cultural and biochemical characteristics, the isolates grouped into nine morphotypes indicating the diverse nature of rhizobia nodulating with MAC beans. Cluster analysis based on genetic characteristics obtained after PCR-ARDRA profiling, showed a larger proportion of significant (JK, OO, JM, and EN conceived and designed the research and data collection tools and participated in drafting the manuscript. GK and SM collected the data, participated in data analyses, and wrote the manuscript. EN performed the data analyses. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Sertoli cells (Sc) are unique somatic cells of testis that are the target of both FSH and testosterone (T) and regulate spermatogenesis. Although Sc of neonatal rat testes are exposed to high levels of FSH and T, robust differentiation of spermatogonial cells becomes conspicuous only after 11-days of postnatal age. We have demonstrated earlier that a developmental switch in terms of hormonal responsiveness occurs in rat Sc at around 12 days of postnatal age during the rapid transition of spermatogonia A to B. Therefore, such “functional maturation” of Sc, during pubertal development becomes prerequisite for the onset of spermatogenesis. However, a conspicuous difference in robust hormone (both T and FSH) induced gene expression during the different phases of Sc maturation restricts our understanding about molecular events necessary for the spermatogenic onset and maintenance. Here, using microarray technology, we for the first time have compared the differential transcriptional profile of Sc isolated and cultured from immature (5 days old), maturing (12 days old) and mature (60 days old) rat testes. Our data revealed that immature Sc express genes involved in cellular growth, metabolism, chemokines, cell division, MAPK and Wnt pathways, while mature Sc are more specialized expressing genes involved in glucose metabolism, phagocytosis, insulin signaling and cytoskeleton structuring. Taken together, this differential transcriptome data provide an important resource to reveal the molecular network of Sc maturation which is necessary to govern male germ cell differentiation, hence, will improve our current understanding of the etiology of some forms of idiopathic male infertility. Spermatogenesis is a complex process where every step of male Germ cell (Gc) development are essentially supported by somatic Sertoli cells (Sc) . In respIn rodents, upto 3–5 days after birth, Sc keep proliferating and attract the gonocytes towards the basement membrane for establishing the spermatogonial stem cell (SSC) niche . NeonataTo understand the regulatory mechanism of spermatogenesis, it is imperative to explore the changing landscape of gene expression during various phases of Sc maturation. Microarray is a powerful technique to study differentially expressed genes in large numbers . DiffereRattus norvegicus) were procured from colony maintained by Small Animal Facility, National Institute of Immunology, New Delhi, India. Animals were maintained in a standard day night cycle with stable temperature and humidity and provided food and water ad libitum. All the animal experimentations were approved by Institutional Animal Ethics Committee (IAEC) and performed following standard guidelines of ‘‘Committee for the Purpose of Control and Supervision of Experiment on Animals (CPCSEA),” Government of India.Wistar outbred rats (In vitro hormone treatment was performed on day 4.Sc cultures were prepared from 5 days and 12 days old rat testes as described earlier . BrieflySc from mature rat testes (60 days old) were isolated and cultured as described earlier . BrieflyOn day 4, Gc contamination were found to be less than 5% in Sc cultures of all age groups. Purity of Sc in culture were identified by vimentin staining  whereas Peritubular Cell (PTc) or Leydig Cell (Lc) contaminations were determined by the alkaline phosphatase or the 3β-HSD activity respectively, as described by us before .−7M T in combination = FT media) in pulsatile manner (30min of FT pulse/3hr)The doses of both o-FSH and T were previously found bioactive in cultured Sc obtained from all three age groups studied ; 2) genes downregulated in 12d but upregulated in 60d , and 3) genes upregulated in both 12d and 60d . For each sample, the calculated quantity of each gene was normalized with the endogenous control Ppia (Cyclophilin A). The relative quantities of mRNAs for all target genes were determined by 2-ΔΔCt method as reported by us before [In order to authenticate the array data the expression patterns of some of the genes were validated by quantitative reverse transcription PCR (q-RT-PCR) with additional sets of Sc culture obtained from all three age groups with consistent hormonal treatments (pulsatile FSH and T in combination for 11hr) and were compared with that of the array data. The genes considered were divided in five groups: 1) FSH responsive genes like SCF, GDNF, ABP, Transferrin and Dmrt1 in both 5d and 12d as reported earlier either by us or others . T respos before ,19. It iPostnatal maturation of testicular Sc is critical in regulating male fertility. Therefore, it becomes essential to investigate the genes expressed by Sc during this phase of development. Our microarray results have indicated differential expression of genes associated with cyto-architecture, metabolism, cytokines, chemokines, growth factors, MAPK signaling and Wnt signaling among others. Taken together, this differential transcriptome data provide an important resource database to reveal the molecular network of Sc maturation which is necessary to govern male germ cell differentiation. This information will improve our current understanding of the etiology of some forms of idiopathic male infertility with persistent immature Sc even in adulthood.S1 FigThere are 96 genes common between our data and Zimmerman’s data and these genes are mainly involved in signal transduction, cell-cell communication, energy and metabolism and cell growth.(TIF)Click here for additional data file.S2 FigA) Entity based clustering analysis of co-expressed genes identified four clusters of co-expressed genes. (B) Gene Ontology (GO) based functional analysis of genes grouped under four clusters. Functional analysis was performed for molecular function (MF), cellular components (CC) and biological processes (BP).((TIF)Click here for additional data file.S1 Table(XLS)Click here for additional data file.S2 Table(XLS)Click here for additional data file.S3 Table(XLS)Click here for additional data file.S4 Table(XLS)Click here for additional data file.


Introduction: For many years, amblyopia was regarded as a disorder of the visual system in which an organic cause could not be identified. Optical Coherence Tomography opens new horizons in understanding the etiopathology of amblyopia and seems to highlight morphologic anomalies in the retina of the amblyopic eye. Purpose: The objective of this paper is to analyze the macular thickness, optic nerve changes, and choroidal thickness found in patients diagnosed with amblyopia based on trials reported in the literature.Material and methods: This study analyzes 30 clinical trials regarding amblyopia evaluation with Optical Coherence Tomography. The research articles analyzed were published between 2006 - 2016 and were identified on PubMed database.Results: 19 research studies focused on macular and nerve optic changes, 7 on choroidal changes and 6 on retinal changes after occlusion. The results were discussed according to the type of amblyopia, alteration of macular thickness, optic nerve changes, ganglion cell layer changes, and alteration of choroidal thickness. Conclusions: The results are of great variability, and it seems that macula and choroid involvement is more frequently suggested compared with optic nerve involvement.Abbreviations: OCT = Optical Coherence Tomography, RNFL = Retinal Nerve Fiber Layer, GCC = Ganglion Cell Complex, ACD = Anterior Chamber Depth, BCVA = Best Corrected Visual Acuity  The results are often contradictory because, as Kim says, when conducting an OCT in patients with anisometropia it is important to correct the magnification of the device according to refraction and axial length of the eye. Also, results differ depending on the device used [3].Another limitation regarding correct interpretation of OCT in children emerges from the inexistence of international established normative values of macular and retinal nerve fiber layer (RNFL) parameters in children ;- Wang (2012) stated that the average thickness of the foveola is thicker than in normal eyes, but other regions have no significant difference [5];- Wu (2013) showed in his study that hyperopic anisometropic amblyopic eyes have a thicker foveola than the contralateral eyes [6];- Araki (2014) concluded that there are significant differences between amblyopic and fellow eyes that are independent of abnormalities in the visual cortex. He showed that the macular outer retinal thickness (1 and 3 mm regions) is significantly thicker in „lazy eyes” [7].- Demircan (2015) discovered that in patients age 5-42 years the mean central macular thickness in the amblyopic eyes was thicker patients under 12 years [After studying anisometropic and strabic ambliopic patients:9];- The Sydney Childhood Eye Study (2009) affirmed that in children aged 6 to 12 years, central macular thickness may be increased in „lazy eyes”, although it is uncertain if this precedes or follows the development of amblyopia [10]; - Al-Haddad (2011) showed in his study that the mean macular thickness was significantly increased in amblyopic eyes compared with fellow eyes. This difference remained significant in the anisometropic group but not in the strabic group. The mean RNFL thickness was similar in amblyopic and fellow eyes [11];- In 2013 Tugcu claimed that in patients with combined (anisometropic and strabismic) amblyopia and in patients with anisometropic amblyopia the thickness of ganglion cell complex (GCC) was significantly increased in both amblyopic and nonamblyopic eyes compared to healthy patients. The strabismic amblyopia group presented with significant reduction in GCC thickness and increased foveal thickness compared to nonamblyopic eyes. He did not found significant differences in morphological and functional measures among amblyopic groups but he detected significant differences in the retinal morphology of both amblyopic and nonamblyopic eyes compared with healthy control subjects [12];- Rajavi (2014) showed that the average foveal thickness of children with moderate and severe amblyopia was 10 microns thicker compared with the congener eye or the control group. Also, male patients had a mean macular thickness thicker than female patients and there was no correlation between the types of amblyopia and macular morphology changes [13]. - Szigeti (2014) revealed that there are subtle changes that affect the outer nuclear layer of the fovea suggesting the possible involvement of the photoreceptors [Concerning deprivational amblyopia:2]; - Kim (2013) investigated OCT changes in deprivational amblyopia due to congenital cataract and concluded that the mean macular thickness in amblyopic eyes was not different from the contralateral eye or control group, but noticed a thickening of the nasal RNFL compared with the fellow eye or control group [14]. - Long (2016) stated that 3 months after surgery, unilateral pediatric cataract patients show no significant interocular anatomical parameters differences except for anterior chamber depht (ACD) and that the parameters evaluated by OCT were not significantly correlated with the final BCVA [15-18].Also, there are scientists like Kee (2006) Repka (2009), Firat (2013) and Yakar (2015) who claim the inexistence of differences between amblyopic patients and the control group in their studies regarding macula morphology .Wu (2013) affirmed in his study that hyperopic anisometropic amblyopic eyes have thicker peripapillary RNFL than the contralateral eyes .Some scientist such as Kee (2006), Repka (2009), Huynh (2009), Firat (2013), Yalcin (2014), Demircan (2015) and Yakar (2015) claimed that there are no differences in RNFL and optic nerve disc morphology between amblyopic patients and control groups [• Choroidal thickness20-25]. However, Celik (2016) stated that, in young Turkish adults with hyperopic anisometropic amblyopia, the subfoveal choroidal, ganglion cell complex, macular and RNFL thickness measurements were not statistically significant between hyperopic anisometropic amblyopic eyes and normal fellow eyes [25].Morphological changes of the choroid evidenced with the help of OCT were studied by some researchers who certified that in amblyopic patients the subfoveolar area is significantly higher than in the congener eye or control group [• Are there morphologic changes after occlusion?26-30]. 6 studies were identified regarding the effect of optical correction and occlusion on the retina, optic nerve and choroid morphology. Oner (2016) analyzed central choroidal thickness before and after 6 months of occlusive treatment and reported no thickness reduction of the central choroid after amblyopia treatment. While Miki (2010), Tugcu (2013), and Yassin (2015) did not found differences between macular thickness and RNFL thickness before and after occlusion therapy, Pang (2011) stated that the central macular thickness was reduced after treatment compared with peripheric macular thickness in which changes were not noticed [31].Also, Liu (2010) stated that some children with various types of amblyopia who failed to achieve normal visual acuity after treatment show macular abnormality on OCT examination [Today, with the help of OCT, a new controversy regarding the presence of morphologic retinal changes in patients diagnosed with amblyopia arises. This opens new horizons in better defining amblyopia and can confirm if ocular structure are affected or not. In the last years, scientists have been trying to determine if morphologic changes in the macula, optic nerve, or choroid of amblyopic patients exist. The results are of great variability and, it seems that macula and choroid involvement is more frequently suggested compared with optic nerve involvement.DisclosureNone.


Transthoracic echocardiography (TTE) and focused cardiac ultrasound (FCU) are now considered essential skills and a requirement of training for physicians working in the intensive care unit (ICU) . TTE is Out of 114 ICUs, 69 (61 %) completed the survey, including 23 (33 %) that admitted cardiothoracic surgical patients and 44 (67 %) that did not. The proportion of ICUs performing TTE (94 %) and LU was high (83 %) but the use of TOE was low (33 %). The proportion of ICU consultants performing TTE, LU, and TOE was lower , respectively. The level of expertise (diagnostic versus focused) was highest in TTE (32 %) compared with TOE (22 %) and LU (12 %).The proportions of intensivists untrained in TTE and LU was 41 % and 30 %, respectively. Perceived barriers included lack of organized training (38 %) and time for training (25 %). Other barriers included a perceived lack of need for training (18 %), insufficient equipment (14 %), and resistance from other ultrasound providers (4 %). The most commonly reported training programs were tertiary courses, such as provided by the Australasian Society of Ultrasound in Medicine (68 %) and University of Melbourne (59 %), rather than board examinations or hands-on workshops.We conclude that although TTE and LU are used frequently in Australasian teaching ICUs, many ICU physicians are yet to be trained due to lack of ICU training programs and time for training. Although tertiary courses are popular and provide training to diagnostic level, they are lengthy and depend on trainers and patient caseload and are not, therefore, scalable. An attractive alternative is to begin training in medical school and to train more physicians in basic ultrasound with shorter, more efficient, and hands-on courses utilizing the internet and ultrasound simulators , advanci


Objective: The study aims to assess the spectrum of cognitive and behavioural disorders in patients affected by Amyotrophic Lateral Sclerosis (ALS) according to the recent consensus criteria [9]. The study also intends to assess the impact of physical disability on cognitive and behavioural abnormalities.Methods: Detailed neurological, neuropsychological and neurobehavioral evaluations were administered to 23 ALS patients, 11 Lower Motor Neuron Disease (LMND) patients and 39 healthy controls. Strong et al.’s criteria [9] were applied to diagnose the presence of cognitive/behavioural impairment. Clinical and neuropsychological scores were used for group comparisons and correlation analyses.Results: In comparison with LMND and controls, a subgroup of ALS patients (∼30%) manifested executive dysfunction, which was severe enough to classify them as cognitively impaired. Action naming difficulties and short-term memory deficits were also observed. Aspontaneity, disorganization and mental rigidity reached clinical relevance in 20% of ALS patients. A small percentage of ALS patients (13%) also had comorbid dementia. The cognitive or behavioural status was not related to the clinical features of ALS.Conclusion: The use of consensus criteria for cognitive and behavioural impairment and the comparison with the LMND group proved useful in defining the spectrum of non-motor manifestations of ALS.


Although multiple serum antiretinal autoantibodies (ARAs) have been reported in patients with paraneoplastic and non-paraneoplastic autoimmune retinopathy ((n)pAIR), not all retinal antigens involved in (n)pAIR are specified. This study aims to serologically identify patients with presumed (n)pAIR through determination of both known and unknown ARAs by autoantibody profiling.An antigen suspension bead array using 188 different antigens representing 97 ocular proteins was performed to detect ARAs in serum samples of patients with presumed (n)pAIR (n = 24), uveitis (n = 151) and cataract (n = 21). Logistic regressions were used to estimate the associations between ocular antigens and diagnosis. Validation of interphotoreceptor matrix proteoglycan 2 (IMPG2) and recoverin antigens was performed by immunohistochemistry and immunoblot, respectively.Samples of patients with presumed (n)pAIR exhibited a broad spectrum of ARAs. We identified retinal antigens that have already been described previously (e.g. recoverin), but also identified novel ARA targets. Most ARAs were not specific for (n)pAIR since their presence was also observed in patients with cataract or uveitis. High titers of autoantibodies directed against photoreceptor-specific nuclear receptor and retinol-binding protein 3 were more common in patients with presumed (n)pAIR compared to uveitis . The presence of all other ARAs did not significantly differ between groups. In patients with presumed (n)pAIR, anti-recoverin autoantibodies were the most prevalent ARAs. Validation of bead array results by immunohistochemistry (anti-IMPG2) and immunoblot (anti-recoverin) showed concordant results in (n)pAIR patients.Patients with (n)pAIR are characterized by the presence of a broad spectrum of ARAs. The diagnosis of (n)pAIR cannot be based on the mere presence of serum ARAs, as these are also commonly present in uveitis as well as in age-related cataract patients. Paraneoplastic and non-paraneoplastic autoimmune retinopathy ((n)pAIR) is a rare blinding retinal disorder of unknown pathogenesis. It is presumed that antiretinal autoantibodies (ARAs) are involved in the pathogenesis of (n)pAIR and damage ocular tissue causing poor visual outcome. Symptoms associated with (n)pAIR are progressive visual loss , visual field loss frequently associated with a ring scotoma or loss of the peripheral field, and decreased amplitudes on electroretinogram (ERG). –4Paraneoplastic autoimmune retinopathy (pAIR) includes two subgroups: cancer associated retinopathy (CAR) and melanoma associated retinopathy (MAR). In pAIR the presence of the same auto-antigens in both retinal tissue and malignant tissue has previously been described (e.g. recoverin). –7 The prMultiple serum ARAs have regularly been reported in affected patients , althougCurrently, antigen bead arrays are being used to profile autoantibody reactivity in body fluids. With thiOur study aimed to serologically identify patients with presumed (n)pAIR through determination of ARAs. For this purpose, we used a bead array-based multiplex assay for autoantibody profiling using 188 ocular antigens representing 97 different retinal proteins.Serum samples were either collected during routine diagnostic analysis for the presence of anti-recoverin autoantibodies in the Laboratory of Medical Immunology of the Erasmus University Medical Center between April 2013 and August 2015 or were obtained from biobank of our department. The study was approved by the local ethical committee from the Erasmus University Medical Center  and adhered to the tenets of the Declaration of Helsinki. The ethical committee decided that no informed consent of patients was required for the use of the remainder of the diagnostic material, as the samples were anonymized and the patients were not subjected to additional risk or procedures. Samples which were obtained from the biobank  included signed informed consent from all participants. All whole blood samples were centrifuged after at least 30’ clotting time at 3,000 rpm for 10 minutes, and serum was stored at -80°C.According to the recently published report on the nomenclature of (n)pAIR, the general term autoimmune retinopathy (AIR) is recommended to indicate the non-paraneoplastic autoimmune retinopathy (npAIR) subtype. In our present series we indicate the specific subtype(s) of AIR pAIR) to prevent any misunderstanding regarding nomenclature. The diagAn additional group of presumed pAIR (CAR or MAR) patients (N = 7) in whom ERG or visual field tests were not performed , but who fulfilled all other inclusion criteria, was included separately. An additional required criterion for these patients comprised the development of a malignancy before or within 3 months after presentation with ocular problems.We collected various clinical data of the patients with presumed (n)pAIR, including patient demographics (age and gender) and ocular characteristics such as complaints of photopsia, complaints of nyctalopia, subjective or objective problems with colour-vision, unilateral or bilateral visual problems and the presence of a malignancy in the medical history or during follow-up.Controls consisted of two groups: 21 serum samples from cataract patients without retinal damage and 151 samples from patients with uveitis of different causes. Patients with age related cataract were included as controls rather than healthy people, as this disorders does not involve retina nor exhibits retinal damage, and represents a clinical setting in which the tests might be employed. Samples of control patients were collected at the department of Ophthalmology of the Erasmus University Medical Center between February 2009 and April 2015. Patient demographics (age and gender) and known malignancies of these patients were registered.Escherichia coli, with an affinity tag consisting of six histidines and an albumin binding domain from streptococcal protein G (His6ABP)  and one bead identity went through the coupling process without addition of antigen . After incubation, the coupled beads were washed and stored in a blocking reagent before combining all bead identities to create a bead array in suspension. Samples were distributed across 96-well microtiter plates, together with triplicate aliquots of a sample pool and a buffer blank in each plate for determination of the intra- and inter-reproducibility. Serum samples were diluted 1:250 in assay buffer before being mixed with the bead array. Incubation was performed at room temperature for 2 hours followed by detection of the IgG reactivity by a fluorophore conjugated anti-human IgG Fab fragment and measured in a FlexMap3D instrument (Luminex Corp.).Autoantibody profiling was performed in all serum samples from patients with presumed (n)pAIR (n = 24), uveitis (n = 151) and cataract (n = 21). Antigens used for the autoantibody profiling were selected based on potential relevance to ocular diseases according to literature and previous positive retinal immunohistochemistry staining, resulting in 188 antigens (human protein fragments) representing 97 unique proteins. The protein fragments were produced within the Human Protein Atlas and designed to represent unique parts of each target protein., 43 ProtHis6ABP) . ImmobilFor validation purposes, samples that tested positive for anti-recoverin autoantibodies on the antigen bead array, and all samples from patients with presumed (n)pAIR, were analysed on a recoverin specific immunoblot . Membrane strips coated with recombinant human recoverin were incubated with a sample buffer for 5 minutes. After aspiration of the sample buffer, the membrane strips were incubated with diluted serum samples for 30 minutes on a shaking platform. Subsequently membrane strips were washed three times, incubated with secondary antibodies (enzyme conjugated anti-human IgG), washed again for three times and stained with a substrate solution which was capable of promoting an enzymatic colour reaction. To identify positive reactions, assessment of visible bands was performed relative to the included control. Results from the antigen suspension bead array and the recoverin specific immunoblot were compared.® , followed by hydration in graded alcohols and blocking for endogenous peroxidase in 0.3% hydrogen peroxide. For antigen retrieval, slides were immersed and boiled in Citrate buffer®, pH6  for 4 min at 125°C and then allowed to cool to 90°C. Automated immunohistochemistry was performed essentially as previously described, using an Autostainer 480 instrument® (Lab Vision). Ant Ant46] AOur study shows that patients with (n)pAIR are characterized by the presence of a broad spectrum of various ARAs. We identified ARAs that have already been described in previous studies, such as anti-recoverin autoantibodies, but also identified new retinal targets. Our findings illustrate that serum ARAs are not only present in patients with (n)pAIR, but also in patients with cataract and uveitis. Though some ARAs appeared to be specific for (n)pAIR, their prevalence and consequently their sensitivity as markers for (n)pAIR were low. This autoantibody screening using 188 antigen provides insight into the autoimmune repertoire of patients with (n)pAIR and a base for further validation with independent methods for protein analysis and independent sample cohorts.A gold standard for the determination of ARAs is currently lacking. DifferenIn addition, information on clinical relevance of the specific ARAs and their pathological titres are lacking. A combination of different ARAs was observed in some cases and therefore their individual effects on retinal tissue could not be distinguished.In our study, we performed statistical analyses using different cut-off levels. By using a high cut-off value, a ratio > 25, false positive results were minimized and retinal targets with a high specificity for (n)pAIR were found. The low cut-off value, a ratio of >2 (indicating ARAs with at least twice the reactivity of the negative control), was used for a more sensitive approach, limiting the exclusion of possibly relevant ARA targets with a lower titre. However, with both cut-off values, no ARAs were found eligible for diagnostic purposes. Some ARAs were specific for (n)pAIR, but had low prevalence while others were more frequently identified but lacked specificity.In concordance with previous findings, positive results of serum anti-recoverin autoantibodies were not only observed in patients with presumed (n)pAIR, but also in patients with uveitis and cataract. Furthermore, no association between the presence of autoantibodies directed against recoverin and the presence of a malignancy was found. A discrepancy between the antigen suspension bead array results for anti-recoverin and the anti-recoverin immunoblot was found in three control patients (one with cataract and two with uveitis). The difference in results could be explained by the different techniques used for determination of ARAs imposing differences in analytical performance. Possibly, the number and/or availability of recoverin antigenic epitopes differed between the antigen suspension bead array and the immunoblot technique. The protein fragments we used in this study to screen for autoantibody reactivity in serum were designed to represent unique parts of each target protein. The binding of the autoantibodies towards their target may be influenced by the protein folding of antigens and may differ in comparison to full-length protein arrays or peptide arrays. Both linear and conformational epitopes, recognized by some ARAs, might be missed for some proteins, preventing recognition by certain autoantibodies.The identification of new ARAs in (n)pAIR is in line with findings from previous studies using Western blot analysis for the determination of ARAs., 48 AlthOur present study focused on the autoantibodies prevalent in serum, which reflects systemic production and is probably not influenced by potential  production or accumulation of specific autoantibodies within the eye. Analysis of local, intraocular retinal autoantibodies might show an entirely different pattern and may differ in clinical importance compared to retinal autoantibodies found in the peripheral circulation. The importance of locally produced autoantibodies has already been shown in cerebrospinal fluid for the central nervous system. In addition, it is unknown which autoantibodies penetrate from the circulation, through the blood retina barrier, into the retina and cause a local inflammation. Further research addressing the intraocular presence of specific retinal autoantibodies might elucidate the clinically relevant autoimmune processes directed against the retinal tissue in (n)pAIR.A gold standard for the definitive diagnosis of (n)pAIR is currently lacking. Also in this study, the diagnosis of presumed (n)pAIR was based on clinical symptoms. To compensate for this inaccuracy, we used strict inclusion criteria and selected a uniform cohort of patients with unexplained visual loss, visual field defects and decreased or absent ERG while other diagnostic possibilities leading to this configuration of clinical characteristics were (so far as possible) excluded. The presence of ARAs was found in all our patients with presumed (n)pAIR and therefore all fulfilled the criteria for the diagnosis of (n)pAIR.Although the mere presence of ARAs supports the diagnosis of (n)pAIR, it has been stated that there are no specific ARAs which would be exclusive for (n)pAIR and none of the ARAs were identified to be of higher diagnostic value than other ARAs., 9, 14 OIn conclusion, our study identified a heterogenous reactivity pattern of ARAs in serum of patients with (n)pAIR, although the presence of ARAs was not discriminatory between (n)pAIR, cataract and uveitis and exhibited a low sensitivity. Therefore, the diagnosis of (n)pAIR cannot be based on the mere presence of serum ARAs and such presence thus warrants careful interpretation. The determination of ARAs in intraocular fluid might provide more insight into the pathogenesis of (n)pAIR and might indicate more sensitive and specific diagnostic tools. Therefore, future research on the prevalence of ARAs in ocular fluid represents an important next step.S1 Table(DOCX)Click here for additional data file.S1 FigARAs not prevalent in patients suspected of paraneoplastic and non-paraneoplastic autoimmune retinopathy are not shown.* GNAS;GNAZ;GNA11-15;GNAL;GNAQ;GNAI1-I3;GNAO1;GNAT1-T3.(EPS)Click here for additional data file.


Gadus morhua) and turbot  in Denmark. Phylogenomic analyses revealed that these two ranaviruses are nearly identical and form a distinct clade at the base of the ranavirus tree branching off near other fish ranaviruses.Ranaviruses have been isolated from Atlantic cod ( Cod iridovirus (CoIV) is the earliest isolation of a ranavirus in fish. It was isolated from wild Atlantic cod in Danish coastal waters in 1979 during an investigation of ulcers in cod . Ranavirg for 20 min, and total nucleic acids were purified using a DNeasy blood and tissue kit (Qiagen). The DNA libraries were prepared using the Nextera XT DNA kit (Illumina), and sequencing was performed using a V3 chemistry 600-cycle kit on a MiSeq platform (Illumina). De novo assembly of the paired-end reads in SPAdes (The Rmax and CoIV isolates were propagated in bluegill fry (BF-2) cells as previously described  and harvn SPAdes  producedIridoviridae core genes (The genomes of Rmax and CoIV were annotated using GATU  with epire genes  revealedThe isolation of nearly identical ranavirus strains from two different marine species, hundreds of kilometers and 20 years apart, indicates the persistence of a ranavirus strain of unknown virulence in the marine environment. With the exception of the grouper iridoviruses in Asia , the impKX574343 and KX574342, respectively.The complete genome sequences of Rmax and CoIV have been deposited in GenBank under the accession numbers


This article provides original data on median dietary intake of 18 amino acids from amino acid medical foods, glycomacropeptide medical foods, and natural foods based on 3-day food records obtained from subjects with phenylketonuria who consumed low-phenylalanine diets in combination with amino acid medical foods and glycomacropeptide medical foods for 3 weeks each in a crossover design. The sample size of 30 subjects included 20 subjects with classical phenylketonuria and 10 with a milder or variant form of phenylketonuria. Results are presented for the Delis-Kaplan Executive Function System and the Cambridge Neuropsychological Test Automated Battery; the tests were administered at the end of each 3-week dietary treatment with amino acid medical foods and glycomacropeptide medical foods. The data are supplemental to our clinical trial, entitled “Glycomacropetide for nutritional management of phenylketonuria: a randomized, controlled, crossover trial, 2016 (1) and “Metabolomic changes demonstrate reduced bioavailability of tyrosine and altered metabolism of tryptophan via the kynurenine pathway with ingestion of medical foods in phenylketonuria, 2017 (2). This data has been made public and has utility to clinicians and researchers due to the following: 1) This provides the first comprehensive report of typical intakes of 18 amino acids from natural foods, as well as amino acid and glycomacropeptide medical foods in adolescents and adults with phenylketonuria; and 2) This is the first evidence of similar standardized neuropsychological testing data in adolescents and adults with early-treated phenylketonuria who consumed amino acid and glycomacropeptide medical foods. Specifications TableValue of the data•The data presented are the first comparison of how ingestion of medical foods comprised primarily of single amino acids or intact protein from glycomacropeptide (a 64-amino acid glycophosphopeptide isolated from cheese whey) affect the dietary intake profile of amino acids.•The dietary intake of 18 amino acids provides useful information to clinicians and researchers related to typical amino acid intake of individuals with phenylketonuria.•The data from the standardized neuropsychological tests can be compared with pharmacological studies using these same tests to contrast the effectiveness of dietary management with pharmacological treatment in subjects with phenylketonuria •These data are useful to clinicians and researchers evaluating the safety and efficacy of glycomacropeptide medical foods in the nutritional management of phenylketonuria.1Summary data for dietary intake of amino acids and assessment of neuropsychological and executive function are presented for subjects with phenylketonuria enrolled in a randomized, controlled, crossover trial conducted from November 2010 to July 2015 2Thirty subjects with early-treated phenylketonuria, 20 with classical and 10 with a milder or variant form of phenylketonuria, completed the clinical trial 3Daily amino acid intake from the whole diet, medical foods, and natural food was calculated from consecutive 3-day food records at the end of the 3-week amino acid medical foods and glycomacropeptide medical foods treatments . Amino aNeuropsychological testing to assess executive function, the Delis-Kaplan Executive Function System™ , and the Cambridge Neuropsychological Test Automated Battery  was conducted by research staff trained in the administration of standard psychological assessments under the supervision of a licensed psychologist after following the amino acid medical foods and glycomacropeptide medical foods treatments for 3 weeks , Table 3Department of Health and Human Services Grants R01 FD003711 from the FDA Office of Orphan Products Development to Ney, P30-HD-03352, T32 DK007665 to Ney, and by the Clinical and Translational Science Award (CTSA) program, through the NIH National Center for Advancing Translational Sciences (NCATS), Grant UL1TR000427. Cambrooke Therapeutics, Inc. donated the glycomacropeptide medical foods used in this study.This work was supported by


The physical examination showed violaceous petechial purpuric patches, coalescing by area, ulcerated nodular lesions extended to both lower limbs, with presence of necrosis in center of lesions (Vasculitis is an inflammatory disease of blood vessels characterized by the alteration or destruction of the vessel wall. A 55 year’s old woman, followed for diabetes and hypertension, presented to our consultation complaining of generalized pruritic lesions, without fever arthritis or any other systemic manifestations. The patient reported the concept of taking Oxicam; Brexin lesions Figure 1A lesions Figure 1B


Resistance to intravenous immunoglobulin (IVIG) occurs in 10–20% of patients with Kawasaki disease (KD). The risk of resistance is about two-fold higher in patients with elevated gamma glutamyl transferase (GGT) levels. We sought to understand the biological mechanisms underlying IVIG resistance in patients with elevated GGT levels.We explored the association between elevated GGT levels and IVIG-resistance with a cohort of 686 KD patients (Cohort I). Gene expression data from 130 children with acute KD (Cohort II) were analyzed using the R square statistic and false discovery analysis to identify genes that were differentially represented in patients with elevated GGT levels with regard to IVIG responsiveness. Two additional KD cohorts (Cohort III and IV) were used to test the hypothesis that sialylation and GGT may be involved in IVIG resistance through neutrophil apoptosis.ST6GALNAC3, a sialyltransferase, as the most differentially expressed gene, we hypothesized that sialylation and GGT are involved in IVIG resistance through neutrophil apoptosis. We then confirmed that in Cohort III and IV there was significantly less reduction in neutrophil count in IVIG non-responders.Thirty-six genes were identified that significantly explained the variations of both GGT levels and IVIG responsiveness in KD patients. After Bonferroni correction, significant associations with IVIG resistance persisted for 12 out of 36 genes among patients with elevated GGT levels and none among patients with normal GGT levels. With the discovery of Gene expression analyses combining molecular and clinical datasets support the hypotheses that: (1) neutrophil apoptosis induced by IVIG may be a mechanism of action of IVIG in KD; (2) changes in sialylation and GGT level in KD patients may contribute synergistically to IVIG resistance through blocking IVIG-induced neutrophil apoptosis. These findings have implications for understanding the mechanism of action in IVIG resistance, and possibly for development of novel therapeutics. Kawasaki disease (KD) is an acute systemic vasculitis of infants and children, that occurs world-wide , but theIn this study, we explored the association of GGT level and IVIG resistance in KD patients to try to understand the biological mechanism of action underlying these two clinical findings. With our systematic analyses of gene regulation and associated clinical findings and outcomes, we hypothesize: (1) neutrophil apoptosis induced by IVIG plays a pivotal role in IVIG responsiveness; (2) changes in sialylation and GGT levels in acute KD patients may contribute together to IVIG resistance by blocking IVIG-induced neutrophil apoptosis.The normal ranges of GGT and alanine aminotransferase (ALT) are shown in Table A in A cohort with 686 subjects with KD (Cohort I) was used to explore the associations between GGT levels and IVIG resistance. All subjects in this cohort were enrolled at Rady Children’s Hospital in San Diego after obtaining written parental informed consent and patient assent as appropriate. All subjects were treated with 2 g/kg of IVIG (Gammagard®) over a 10–12 hour period as per the pharmacy’s standard protocol. The study protocol was conducted in accordance with the Declaration of Helsinki and was approved by Institutional Review Boards of UCSD and Stanford University. KD subjects in this study were: a) patients with fever  for no more than 10 days, plus at least four of the five principal clinical criteria, b) patients meeting fewer criteria but with coronary artery abnormalities (CAA) (Z-score ≥2.5 for left anterior descending [LAD] and/or right coronary arteries [RCA]) documented by echocardiogram, and c) patients with fewer than 4 clinical criteria but meeting the American Heart Association (AHA) criteria for incomplete KD by laboratory criteria [We performed gene expression profiles of whole blood obtained from a sub-cohort of 146 KD subjects (Cohort II) as previously described . Of thesFor each of the 31,000 genes in the 130 subjects, the explained variations in GGT levels or IVIG responsiveness were estimated using R square of linear regression or the McFadden's pseudo- R square , 19 of lCorrection for multiple hypothesis testing on the explained variations was performed using the permutation-based false discovery rate (FDR) analytical method . FDRs ofThe study population was divided into two sub-cohorts, one with normal and the other with elevated GGT levels, using age-specific reference ranges established in our clinical laboratory  with 587 KD subjects. Approved by the institutional review board of Korean University Medical Center, the medical records of KD patients from January 2008 to December 2013 were reviewed. The diagnosis and treatment of KD was based on AHA criteria . CAA werThe absolute neutrophil count (ANC) (mature and immature (band) forms) were calculated for both the UCSD and Korean cohorts.All statistical analyses and plots were performed using R 3.2.2  and ggplThe 575 IVIG responders and 111 IVIG non-responders in Cohort I had similar demographic and clinical characteristics . SubjectIVIG resistance was examined according to GGT levels quintiles . IVIG reOf the 130 KD subjects with gene expression data in Cohort II, 100 were IVIG responders and 30 were IVIG non-responders. At the time of diagnosis, 45 subjects had normal serum GGT levels, and 85 had elevated GGT levels  with IVIG resistance persisted for 12 out of the 36 genes among patients with elevated GGT levels and none among patients with normal GGT levels  or elevated GGT levels (85 subjects including 62 IVIG responders and 23 non-responders). After applying the Bonferroni correction, significant associations  percentage reduction in neutrophil count than responders. A similar pattern was observed in Cohort IV, which included 222 IVIG non-responders and 365 IVIG responders . We further delineated changes in ANC by stratifying patients into quintiles according to the ANC at presentation ..ST6GALNAactoside , 35. Neuin vitro , suggestin vitro . These oin vitro . Circulain vitro , 39.Despite the fact there have been reports of KD occurring in the presence of Autoimmune neutropenia (AIN), 41, forST6GALNAC3 leads to increased enzymatic activity of ST6GALNAC3, producing more GD1α, which in turn binds to siglec-9 and prevents its recognition by Nabs-siglec-9 in IVIG; (2) elevation of GGT levels will increase the degradation of extracellular glutathione (GSH) to provide cysteine for de novo synthesis of intracellular GSH, reducing the intracellular reactive oxygen species (ROS) in neutrophils. Reduction of ROS in neutrophils decreases neutrophil apoptosis induced by IVIG in the presence of GM-CSF and IFN- γ [Naturally occurring antibodies (Nabs) against siglec-9 (Nabs-siglec-9) are found in IVIG and can induce apoptosis in neutrophils –44. MoreTo the best of our knowledge, we are the first to integrate multifaceted data sets of expression profiles, clinical parameters and outcomes to explore KD pathophysiology. We demonstrated that KD subjects with elevated GGT levels have a unique gene expression pattern that overlaps with the gene expression pattern associated with IVIG resistance. Our study suggests that reduction of circulating neutrophils is one of the hallmarks of the therapeutic effects of IVIG.ST6GALNAC3 and elevated GGT levels may lead to reduced neutrophil apoptosis, and consequently IVIG resistance  Nabs against siglec-9 (Nabs-siglec-9) in IVIG can induce apoptosis in neutrophils and contribute to the efficacy of IVIG in the treatment of KD. (2) Increased sialylation of gangliosides block IVIG-mediated neutrophil apoptosis and lead to persistent inflammation and IVIG resistance.If confirmed, our findings may account for the variable effectiveness of different IVIG lot preparations , 49, potS1 FigNumbers below the x axis are numbers of subjects whose ANC increased after IVIG treatment. Of the 13 IVIG responders with ANC increased after IVIG, 12 fell into the lowest quintile of acute phase neutrophils, and none in the last three quintiles.(TIF)Click here for additional data file.S2 FigLoess curve analyses of neutrophil reduction or the percentage reduction in neutrophil count as a function of the pretreatment ANC .(TIFF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S1 FileTable A in S1 File. The standard range for normal GGT (IU/L); Table B in S1 File. The standard range for normal ALT (IU/L); Table C in S1 File. The clinical characteristics and laboratory values for subjects selected for gene expression analysis; Table D in S1 File. Association of 12 genes with IVIG response in subgroup with normal and elevated CRP levels; Table E in S1 File. Association of 12 genes with IVIG response in subgroup with normal and elevated CRP levels.(PPTX)Click here for additional data file.


HSPD1 gene that encodes the HSP60 subunit of the HSP60/HSP10 chaperonin complex. Investigations of the molecular mechanisms underlying these disorders have revealed that different degrees of reduced HSP60 function produce distinct neurological phenotypes. While mutations with deleterious or strong dominant negative effects are not compatible with life, HSPD1 gene variations found in the human population impair HSP60 function and depending on the mechanism and degree of HSP60 dys- and mal-function cause different phenotypes. We here summarize the knowledge on the effects of disturbances of the function of the HSP60/HSP10 chaperonin complex by disease-associated mutations.Heat shock protein 60 (HSP60) forms together with heat shock protein 10 (HSP10) double-barrel chaperonin complexes that are essential for folding to the native state of proteins in the mitochondrial matrix space. Two extremely rare monogenic disorders have been described that are caused by missense mutations in the  E. coli bacteria  stacked back to back  ) lists 95 missense variations for the protein product of the canonical HSPD1 transcript. This website provides high quality exome sequencing data from more than 60,000 unrelated adults without severe pediatric disease. Of the 95 HSPD1 missense variations 60 have only been seen in one single allele and only 9 have been observed in more than 10 of the ~120,000 alleles recorded , this variation may be disease-associated with reduced penetrance. The most frequently observed amino acid variation in HSP60, HSP60-p.Gly563Ala, has been found in homozygous form in one sporadic Danish spastic paraplegia patient  has been reported (Hansen et al., Effects of expressing the SPG13- and MitCHap60-associated mutant proteins on mitochondrial morphology have been assessed in Cos-7 cells transfected with cDNAs encoding the disease-associated mutant proteins (Miyamoto et al., ShRNA-mediated knock-down of HSP60 in human HEK293 cells decreased the steady state levels of the mitochondrial medium-chain acyl-CoA dehydrogenase (Corydon et al., Knock-out of both HSP60-encoding alleles in mice is not compatible with life. Such embryos died early during development (Christensen et al., E. coli 85 proteins that display obligate dependence on folding assistance by the bacterial chaperonin complex have been characterized (Kerner et al., Proteins like UQCRC1 or SOD2 thus appear to depend more than others proteins on folding assistance by the HSP60/HSP10 complex. For de novo mutations in the HSPD1 and HSPE1 genes in sporadic diseases. Clinical geneticists should therefore be aware of this and it will be important to collect the knowledge of these rare cases to be able study genotype/phenotype relationships and to assess the disease-causing nature of variations.Different mutations in HSP60 or its partner protein HSP10 lead to distinct phenotypes of neurological disorders with a clear mitochondrial dysfunction pattern. These diseases are very rare as deleterious effects of mutations in these essential genes are not compatible with normal embryonal development (Christensen et al., Besides being affected by mutations, the activity and function of the HSP60/HSP10 complex can also be regulated by its expression levels. The regulation of the transcription levels of both proteins occurs via different elements in the bidirectional promoter. SP1 elements provide robust house-keeping levels of expression and on top of that heat-shock elements, mitochondrial unfolding protein response elements, and STAT3 elements further modulate expression adapting it to specific situations (Zhao et al., Indeed, dysregulation of HSP60 expression in hypothalamus has been implicated with type 2 diabetes mellitus (Kleinridders et al., Finally, posttranslational modifications of the HSP60/HSP10 complex may regulate the activity of the complex. Like other molecular chaperones HSP60 is a highly modified protein with a long list of PTMs recorded in the UniProt database (Consortium, PF and PB have both contributed to writing the draft and producing the final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Non-traumatic out of hospital cardiac arrest (OHCA) is the leading cause of death worldwide, acute coronary syndromes accounting for up to 2/3 of cases. Urgent angiography with a view to primary percutaneous coronary intervention (PPCI) is a class IB recommendation according to international guidelines. Diagnosis and management of patients with unobstructed coronaries or unidentified culprit lesion on angiogram is challenging. Cardiac magnetic resonance (CMR) with its superior tissue characterization is a well-established diagnostic technique. We sought to assess the additive role of CMR in diagnosis and management of patients with an inconclusive coronary angiogram and to determine which findings on CMR best predict clinical impact.We retrospectively analysed our database to collect data on consecutive patients surviving non-traumatic OHCA, undergoing urgent coronary angiogram and CMR. We focused the analysis on patients with an inconclusive angiogram, defined as the evidence of unobstructed coronaries or of coronary artery disease (CAD) without a clear culprit lesion. Clinical impact of CMR was defined as a change in diagnosis, as compared to that made on a multi-parametric pre-CMR basis , or a change in management, which could be a change in medication or the performance/avoidance of invasive procedures .Out of 157 patients surviving OHCA referred for CMR after urgent angiogram, we identified 104 patients  with inconclusive angiogram (66%): 67 patients (64%) had unobstructed coronaries and 37 (36%) had CAD with no clear culprit. Diagnosis based on CMR findings was ischemic heart disease in 42 patients (40%), non-ischemic heart disease in 30 (29%), a structurally normal heart was found in 25 patients (24%) and non-specific findings in 7 (7%). Overall, CMR had a clinical impact in 68/104 patients (65%), determining a change in diagnosis in 17% of patients, a change in management in 31% and both a change in diagnosis and management in 17%. CMR led to myocardial revascularization in 18% of patients and to ICD implantation in 16%; based on CMR findings, an invasive procedure was avoided in 17% of patients. In a multivariate model that included clinical and imaging parameters, LGE  and segmental regional wall motion abnormality  were the strongest independent predictors of CMR clinical impact (Table CMR had an additive clinical impact on diagnosis and management in 65% of patients surviving OHCA with an inconclusive coronary angiogram. LGE and segmental regional wall motion abnormality were the best independent predictors of clinical impact following CMR. CMR should be enclosed in the clinical-diagnostic work-up of this subgroup of OHCA survivors.


Scientific Reports 10.1038/s41598-017-14674-5, published online 06 November 2017Correction to: The original version of this Article contained an error in the Abstract.“Changes in miRNA expression glomerular of capillaries during antibody-mediated rejection (ABMR) are poorly understood and could contribute to the deleterious inflammation and fibrosis of ABMR via suppression of target genes.”now reads:“Changes in miRNA expression of glomerular capillaries during antibody-mediated rejection (ABMR) are poorly understood and could contribute to the deleterious inflammation and fibrosis of ABMR via suppression of target genes.”This error has now been corrected in the PDF and HTML versions of the Article.