A series of intensive, longitudinal, mark-recapture studies of hantavirus infection dynamics in reservoir populations in the southwestern United States indicates consistent patterns as well as important differences among sites and host-virus associations. All studies found a higher prevalence of infection in older  mice; one study associated wounds with seropositivity. These findings are consistent with horizontal transmission and transmission through fighting between adult male rodents. Despite very low rodent densities at some sites, low-level hantavirus infection continued, perhaps because of persistent infection in a few long-lived rodents or periodic reintroduction of virus from neighboring populations. Prevalence of hantavirus antibody showed seasonal and multiyear patterns that suggested a delayed density-dependent relationship between prevalence and population density. Clear differences in population dynamics and patterns of infection among sites, sampling periods, and host species underscore the importance of replication and continuity of long-term reservoir studies. Nevertheless, the measurable associations between environmental variables, reservoir population density, rates of virus transmission, and prevalence of infection in host populations may improve our capacity to model processes influencing infection and predict increased risk for hantavirus transmission to humans.


Although basic mechanisms of bronchial hyper-responsiveness (BHR)are still incompletely understood, inflammation of airways is likelyto play a fundamental role in modulating BHR in patients withasthma. The involvement of several inflammatory cells  andof bioactive mediators secreted by these cells in the pathogenesisof asthma is well documented. Sodium cromoglycate and nedocromilsodium are two pharmacological agents which have anti-allergic andanti-inflammatory properties. Their clinical effectiveness in mildto moderate asthma, and the capacity to reduce BHR under differentnatural and experimental conditions, make them valuable drugs formaintenance therapy in patients with asthma.


Beyond the classic Normal Pressure Hydrocephalus (NPH) triad of gait disturbance, incontinence, and dementia are characteristic signs of motor dysfunction in NPH patients. We used highly sensitive and objective methods to characterize upper limb extrapyramidal signs in a series of NPH subjects compared with controls. Concentrated evaluation of these profound, yet underappreciated movement disorders of NPH before and after techniques of therapeutic intervention may lead to improved diagnosis, insight into pathophysiology, and targeted treatment.Twenty-two (22) consecutive NPH patients and 17 controls performed an upper limb motor task battery where highly sensitive and objective measures of akinesia/bradykinesia, tone, and tremor were conducted. NPH subjects performed this test battery before and more than 36 h after continuous CSF drainage via a spinal catheter over 72 h and, in those subjects undergoing permanent ventriculo-peritoneal shunt placement, at least 12 weeks later. Control subjects performed the task battery at the same dates as the NPH subjects. Statistical analyses were applied to group populations of NPH and control subjects and repeated measures for within subject performance.Twenty (20) NPH subjects remained in the study following CSF drainage as did 14 controls. NPH subjects demonstrated akinesia/bradykinesia (prolonged reaction and movement times) and increased resting tone compared with controls. Furthermore, the NPH group demonstrated increased difficulty with self-initiated tasks compared with stimulus-initiated tasks. Following CSF drainage, some NPH subjects demonstrated reduced movement times with greater improvement in self- versus stimulus-initiated tasks. Group reaction time was unchanged. Resting tremor present in one NPH subject resolved following shunt placement. Tone measures were consistent for all subjects throughout the study.Clinical motor signs of NPH subjects extend beyond gait deficits and include extrapyramidal manifestations of bradykinesia, akinesia, rigidity, and propensity to perform more poorly when external cues to move are absent. Objective improvement of some but not all of these features was seen following temporary or permanent CSF diversion. The classic clinical triad of Normal Pressure Hydrocephalus (NPH) includes gait disturbance, incontinence, and dementia though most patients exhibit less than three of these signs early in the progressive clinical course ,2. In adThat ventriculo-peritoneal shunting reverses each of the classic signs in some NPH patients suggests a causative role in CSF dynamics, but the mechanisms underlying the neuronal and glial pathophysiology of NPH remain obscure. During the course of NPH, periods of elevated intracranial CSF pressure are hypothesized to result in dysfunction of white and gray matter of the brain  . These hFew investigations of NPH patients have included objective and sensitive analysis of extrapyramidal motor signs, and examination of these signs outside of gait dysfunction are rarer still ,10-12. WThis study, together with patient recruitment, was approved by the Johns Hopkins University Institutional Review Board. Signed, informed consent was obtained from each subject. Twenty-two (22) consecutive subjects above the age of 50  referred for evaluation of possible NPH were recruited from the Johns Hopkins University Outpatient Center, with no enrollment restrictions based on race or ethnic origin. Requirements for inclusion were radiographic evidence of ventriculomegaly by CT scan or conventional MRI, and clinical signs of NPH that included gait impairment, urinary incontinence, or cognitive impairment. Subjects were excluded if they had undergone prior shunt or intracranial surgery, were unable to undergo lumbar puncture for CSF drainage or had other active neurologic disease . Controls (n = 17) were partners of eligible NPH subjects  and were required to be free from clinical signs of NPH.NPH subjects were tested with the entire battery of tasks at 1) baseline, 2) within 36 h following removal of approximately 10 ml CSF/h for 72 h via a temporary spinal catheter and 3) approximately 12 weeks following ventriculo-peritoneal shunt insertion (Medtronic). Control subjects were tested with the entire battery of tasks on the same days of testing as NPH subjects. Two NPH subjects demonstrated signs of infection following CSF drainage via spinal catheter and were excluded from further testing and analysis. One subject withdrew consent to participate in the study after baseline testing such that subsequent data were not collected.Subjects performed ballistic movements using their dominant hand by moving a manipulandum that constrained two-dimensional movement about the wrist. Targets, 10° wide and unbounded , were separated by 50°. Practice trials were performed for this simple task to avoid any potential learning curve barriers. Subjects reacted and moved in response to an auditory "GO" signal (stimulus-initiated tasks) or performed the identical movement task in the absence of a "GO" signal (self-initiated tasks). All subjects were informed ahead of time if they were to perform stimulus- or self-initiated tasks, and in both cases received preparatory auditory cues of "ready" and "set" in the initial stages of the task. In stimulus-initiated tasks, these preparatory cues preceded randomly delivered "GO" signals, and in self-initiated tasks, where no "GO" signal was delivered, a minimum time of 2 s of hold time was required after the "set" cue before initiating movement. Errors were recorded and the task discarded if movement occurred prior to 150 ms more than "GO" signal, more than 1 s after the "GO" signal, or if final target was overshot. An electronic goniometer continuously recorded manipulandum position during the task and was fed into an online recording computerized system  along with auditory cues and electromyographic recordings of agonist and antagonist wrist muscles performing the task. Forty-five correctly performed trials were performed by each of the subjects. Reaction time (RT) was measured as time between "GO" signal and first significant change of the agonist electromyographic record (EMG) over baseline (RT/EMG) and between "GO" signal and movement onset (MO) (RT/MO). Electromechanical delay (EMD) was determined as the time between first agonist EMG and movement onset. Movement time (MT) was defined as the time between movement onset (MO) and achieving final target.k = the slope of force over displacement), or active EMG activity in primary agonists or antagonists about the elbow [Muscle tone was measured by an apparatus that quantifies tone about the elbow. Subjects rested their dominant forearm in a molded and padded cradle mounted on a metal beam. The cradle rotated on low-friction ball bearings and an electronic goniometer measured angular displacement. A load cell  located at the side of the metal beam measured the torque applied by the examiner. The torque plotted against the angle (degrees) represented muscle stiffness, and post-hoc least squares fit analysis was then performed to determine the slope of the contour representing tone about the elbow . The tonhe elbow . Three dhe elbow .Tremor was analyzed using triaxial solid-state accelerometers  that detect frequency (Hz) and amplitude  in μm. Triaxial accelerometers were placed on the distal hand while the subjects' hands were at repose for a period of 120 s. Subjects were then asked to hold their arms outstretched, and tremor analysis was repeated during the next 60 s. All data were fed online to a Pentium based computer and analyzed using Fast Fourier Transforms. Mean tremor amplitude and dominant frequency were recorded while at rest and with outstretched arms.Group statistical analyses for parametric measures were compared using ANOVA tests with Fisher's Least Square Difference Post-Hoc analysis when F scores were significant or Kruskal-Wallis comparisons for non-parametric measures of reaction time. Analyses within individuals were compared using Repeated-measures ANOVA with Fisher's Least Square Difference or Friedman's analysis with Mann-Whitney Post-Hoc analysis.p < 0.01), or as RT/MO  when compared with the control group  . Mean movement times for stimulus-initiated and self-initiated tasks are plotted for each NPH subject at baseline in Figure As a group, NPH subjects demonstrated prolonged MT versus controls Table . At baseThose NPH subjects with fastest movement times (least bradykinetic) were least apt to exhibit discrepancies between self- and stimulus-initiated movements. Furthermore, those NPH subjects with fastest movement times were least apt to exhibit wide intertrial movement time variability as reflected by size of error bars Figure .p < 0.05), but this did not hold true for the group analysis comparing NPH subjects before and after continuous CSF drainage . Those subjects demonstrating significant differences of movement times from baseline following CSF drainage maintained this difference following shunt placement. Furthermore, selective improvement of self- versus stimulus-initiated movement times persisted with shunt placement Figure . Controlp < 0.02, Table p < 0.02, R2 = 0.79). Following CSF drainage and following shunt placement, there were also no significant changes in slope (representing rigidity at the elbow) for the NPH subjects. However, there were qualitative differences in tone hysteresis loops following CSF drainage in NPH subjects including greater incidence in EMG activity and sudden tone perturbations in the NPH group not seen in control subjects.Tone measures at baseline demonstrate increased passive tone about the elbow in NPH subjects compared with control cohorts . Although some subjects improved in both gait and upper extremity measures, this was not uniformly so – such that substantial improvement in gait measures did not always coincide with improvement in ballistic measures. Since none of the subjects improved in measures of tone, improvement in gait did not correlate with this measure. Interestingly, some of those subjects that demonstrated increased upper extremity bradykinesia following CSF drainage also demonstrated worsening bradykinesia in gait testing.In a parallel study, these same subjects underwent formal gait analysis  in NPH subjects. This implies that different pathophysiological substrates underlie each of these motor signs in NPH, and/or that these signs emerge independently. Of these extrapyramidal signs, not all respond equally to CSF drainage. Why would only some symptoms respond to CSF drainage? It is plausible that some structures revert to normal function with reduction of direct shearing forces, while others  are irreversible or require time for tissue recovery or compensatory pathways to develop. In addition, there is always a potential for multiple etiologies to confound the presentation and responsiveness of clinical signs related to NPH, but our selection criteria sought to minimize these factors.It is an interesting finding that some NPH patients demonstrated the classic Parkinsonian motor characteristic of improved movement execution in the presence of external cues. Others have seen a less dramatic benefit of external cues when studying NPH patients during gait analysis ,18. Our NPH subjects demonstrate increased tone compared with controls. This tone is unrelated to spasticity and independent of movement speed. Our measures of tone are highly sensitive, and even a significant increase in tone compared with controls may be subclinical. From what is known of NPH, one would expect a supraspinal mechanism underlying increased tone. Such mechanisms have been proposed for the increased tone seen in Parkinsonism . There wThe underlying motor pathophysiology of NPH signs extends beyond gait and bladder dysfunction with prevalence of extrapyramidal signs. These include even complex Parkinsonian signs of reliance upon external cues for improved motor performance. That NPH subjects demonstrated improvement of some signs in short order following CSF drainage suggests reversible pathophysiologic mechanisms that can improve activities of daily living. However, sensitive and objective motor measures in this sample suggest other motor dysfunction is resistant to immediate correction by CSF drainage and may be recalcitrant or require chronic treatment to see improvement.Dr. Michael A. Williams and Dr. Daniele Rigamonti have received honoraria in the past from Medtronic, Inc. Dr. George Thomas worked at the Adult Hydrocephalus Center at Johns Hopkins when this study was conducted. His salary was supported by a grant from Medtronic, Inc.ASM provided study design, oversight of data acquisition, data analysis, and drafting the manuscript. JH tested each subject, performed data collection and first stage data analysis. GT provided subject recruitment, scheduling and evaluation. REM participated in critical review of the manuscript and in evaluation of study design. TOC participated in provided testing environment and critical analysis on data collection and interpretation. MAW contributed in patient evaluation and categorization and critical manuscript review. DR contributed to study design, patient evaluation and critical manuscript review. All authors have read and approved the final manuscript.


Recent advances in genomics provide genetic information from humans and othermammals  traditionally used as models as wellas new candidates (pigs and cattle). In addition, linked enabling technologies,such as transgenesis and animal cloning, provide innovative ways to design andperform experiments to dissect complex biological systems. Exploitation of genomicinformation overcomes the traditional need to choose naturally occurring models.Thus, investigators can utilize emerging genomic knowledge and tools to createrelevant animal models. This approach is referred to as reverse genetics. In contrastto ‘forward genetics’, in which gene(s) responsible for a particular phenotypeare identified by positional cloning (phenotype to genotype), the ‘reverse genetics’approach determines the function of a gene and predicts the phenotype of acell, tissue, or organism (genotype to phenotype). The convergence of classicaland reverse genetics, along with genomics, provides a working definition of a‘genetic model’ organism (3). The recent construction of phenotypic maps definingquantitative trait loci (QTL) in various domesticated species provides insights intohow allelic variations contribute to phenotypic diversity. Targeted chromosomalregions are characterized by the construction of bacterial artificial chromosome(BAC) contigs to isolate and characterize genes contributing towards phenotypicvariation. Recombineering provides a powerful methodology to harvest geneticinformation responsible for phenotype. Linking recombineering with gene-targetedhomologous recombination, coupled with nuclear transfer (NT) technology canprovide ‘clones’ of genetically modified animals.


Repeated treatment with streptozotocin selected toxin resistant subpopulation of insulin producing tumor RINmS cells, characterized increased level of insulin content and secretion. In the present study RINmS cells were found to have higher glucose sensitivity and insulin response compared with parental RINm cells. In addition, compounds known to induce elevated level of cAMP beta-cells, such as isobutyl methyl xanthine, and forskolin, potentiated glucose-induced insulin secretion of RINmS, but had no effect on the naive parental RINm cells. These experiments suggest that differentiation therapy can be utilized for engineering insulin producing cells with improved defense and secretory mechanisms.Differentiation therapy has been proposed as a new approach to selectively engage the process of tumor cell differentiation during chemotherapy of cancer. Our recent


Molecular epidemiologic studies of infectious diseases rely on pathogen genotype comparisons, which usually yield patterns comprising sets of DNA fragments (DNA fingerprints). We use a highly developed genotyping system, IS6110-based restriction fragment length polymorphism analysis of Mycobacterium tuberculosis, to develop a computational method that automates comparison of large numbers of fingerprints. Because error in fragment length measurements is proportional to fragment length and is positively correlated for fragments within a lane, an align-and-count method that compensates for relative scaling of lanes reliably counts matching fragments between lanes. Results of a two-step method we developed to cluster identical fingerprints agree closely with 5 years of computer-assisted visual matching among 1,335 M. tuberculosis fingerprints. Fully documented and validated methods of automated comparison and clustering will greatly expand the scope of molecular epidemiology.


Tissue sections of frozen biopsy specimens obtained from normal and hyperplastic human lymphoid tissues, 33 cases of non-Hodgkin lymphomas as well as various forms of immunoregulatory disorders (angioimmunoblastic and dermatopathic lymphadenopathy) were analysed in immunofluorescence tests (using red TRITC and green FITC double-labelling). A panel of antisera including well-characterized conventional reagents to immunoglobulin classes, T lymphoid and Ia-like antigens, and monoclonal antibodies was used. In selected cases the results were compared with the observations of membrane-marker staining on viable cells in suspension. the findings show that the immunological methods can give a very accurate analysis of the normal and malignant lymphoid cells, and can provide complementary information to conventional histology. The investigator can choose the reagent combinations which give answers to various specific questions: e.g. antisera to light chains establish the monoclonality of lymphomas, whilst staining combinations for human T and Ia-like antigens are particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful in various immunoregulatory disorders. Monoclonal antibodies will be particularly useful reagents for analysing the tissue distribution of lymphoid subpopulations and ancillary cells in tissue biopsy specimens.


Mycobacterium ulcerans infection, all of whom received rifampicin-based oral antibiotic therapy followed by surgical resection (three patients) or oral antibiotics alone (one patient). Following oral antibiotics for between 4 and 8 weeks, viable M. ulcerans was not detectable by culture in three of the patients, or by histology in a fourth patient from whom no specimen for culture was obtained. All cases spent time in a BU-endemic area in coastal Victoria, Australia. Baseline characteristics, diagnosis, treatment received, and histopathology of resected specimens are detailed in Buruli ulcer (BU) was treated primarily with wide surgical excision until recent studies confirmed the efficacy of oral rifampicin combined with intramuscular streptomycin. Whether all-oral antibiotic regimens will be equally effective is unknown. This report describes four patients with M. ulcerans was confirmed by positive polymerase chain reaction (PCR) and isolation of M. ulcerans by culture from swabs obtained prior to treatment. Three patients had ulcerative lesions  suggest that shorter periods may be effective as suggested for the combination of rifampicin and streptomycin In the four patients we have described here, combination oral antibiotic therapy prior to excision led to the inability to recover Treatment of patients with limited BU prior to surgery using rifampicin-based oral antibiotics resulted in culture-negative resection specimens.Clinical healing is slow despite the microbiological activity of oral antibiotics.Apparent relapses that occur during or after treatment may be due to immunologically driven paradoxical reactions rather than primary treatment failure.M. ulcerans infection followed by delayed surgery appears to simplify management by allowing excision and closure in one step without relapse.Rifampicin-based oral antibiotic therapy for the treatment of


Iodised oil (lipiodol) administered via the hepatic artery localises selectively in primary liver cell cancers (hepatocellular carcinomas or HCCs) for prolonged periods and has been used as a vehicle for cytotoxic agents. Despite clinical use, the mechanism of lipiodol retention by tumours has remained unclear, embolisation of oil droplets in the tumour vasculature being the prevailing hypothesis. We have investigated the role of tumour and endothelial cells in lipiodol retention. Human liver tumour (Hep G2) cells and human umbilical vein endothelial cells in culture were exposed to lipiodol. Light microscopy using selective silver impregnation stains and transmission electron microscopy revealed lipiodol incorporation by both cell types, probably by pinocytosis. This was not associated with cellular injury in terms of cell lysis, cell replication or radio-labelled leucine uptake. Histological analysis of 24 HCCs either surgically resected or discovered incidentally at liver transplantation  revealed vesicles of lipiodol in the cytoplasm of tumour cells and endothelial cells lining tumour vessels. Thus, lipiodol is likely to deliver cytotoxic agents directly into tumour cells and endothelial cells, both in vitro and in vivo. This may also apply to other lipids and to other human tumours. These findings have significant therapeutic implications.


Progressive night blindness and constriction of the visual fields are features of Bietti crystalline corneoretinal dystrophy, but here we report a case with the most probable diagnosis of Bietti crystalline dystrophy and good central visual acuity and severely decreased electroretinogram.The patient was a 28-year-old woman with complaint of decreased vision without night blindness. Her both eyes visual acuity were 20/25 with plano refraction. Fundus examination showed intraretinal crystals distributed in the posterior pole and also midperiphery. Fullfield electroretinogram showed decreased scotopic a and b-wave amplitudes.In our patient central foveal region was relatively intact; and this can explain subnormal visual acuity. Although visual acuity was nearly spared, electroretinogram was extremely affected. Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessive disorder of retinal degeneration characterized by numerous tiny sparkling yellow-white spots at the posterior pole of the fundus. The small glistering crystals can also occur in the corneal limbus and circulating lymphocytes ,2. CliniA 28-year-old Iranian Caucasian woman referred to our clinic with complaints of decreased vision without night blindness and visual acuity of both eyes was 20/25 with plano refraction. Her slit lamp examination revealed clear cornea and clear lens, and 1+ pigment in vitreous. Intraocular pressure was 18 mmHg for right eye and 17 mmHg for left eye. She did not have any history of associated other medical conditions or drug usage.Fundus examination showed intraretinal crystals distributed in the posterior pole and also midperiphery. There was midperipheral RPE and choriocapillaris atrophy, peripheral pigment clumping and retinal scarring . FlouresFullfield ERG showed severely decreased scotopic a and b-wave amplitudes. Moreover, the photopic a and b wave amplitudes were both severely attenuated . Visual Bietti first reported cases of tapetoretinal degeneration characterized by yellow glistering retinal crystals, choroidal sclerosis, and marginal crystalline dystrophy of the cornea. Similar cases with no limbal crystalline deposits have been reported and called Bietti crystalline fundus dystrophy or crystalline retinopathy -8. The pAffected patients presented with visual symptoms from the third decade onward. The most common presenting symptom is a decrease in visual acuity, often accompanied by nyctalopia. In the presented patient, visual acuity was good and central vision had not been affected. This sparing of the central vision until late in the disease is also recently described in European patients with BCD .Patient with RPE atrophy and intraretinal crystals confined to the posterior pole tend to show lesser disturbances in the full-field electroretinogram (ERG) . This isHistologically, BCD shows evidence of advanced panchorioretinal atrophy characterized by generalized loss and sclerosis of the choriocapillaris with crystals and complex lipid inclusions within the choroidal fibroblasts. The abnormal inclusions are similar to those found in circulating lymphocytes, keratocytes, and conjunctival and skin fibroblasts ,2. AccorBietti initially noted similarities with retinitis punctata albescens and fundus albipunctatus. These conditions are considered flecked retina syndromes and do not have the distinctive crystalline deposits in patients with Bietti’s crystalline dystrophy. Several advanced cases of Bietti’s crystalline dystrophy have been misdiagnosed as retinitis pigmentosa, since advanced cases may have severe visual loss, extensive chorioretinal atrophy, pigment deposition, minimal crystals, and nonrecordable ERG. The diagnosis of BCD is based on clinical findings; biomicroscopic and ophthalmoscopic appearance are usually sufficient to make diagnosis . We can


The regulation of cholesterol esterification during cell proliferation was studied. The serum free cholesterol, cholesterol esters and lecithin: cholesterol acyltransferase (LCAT) activity of nude mice with and without pancreatic acinar cell tumours and rats with proliferating tissues were determined. In addition, the apparent activity of acyl-CoA: cholesterol acyltransferase (ACAT) in homogenates of nude mouse tumours and proliferating rat tissues were determined and compared with those of normal nude mouse and rat tissues. Serum cholesterol ester levels were significantly lower in host nude mice with tumours and in rats with regenerating liver, and increased significantly in pregnant rats when compared with respective controls. Circulating LCAT activity levels decreased in host nude mice, in pregnant rats, and in rats with regenerating pancreas and regenerating liver. Apparent ACAT activity levels increased significantly in nude mouse tumours and in foetal and postnatal rat pancreata and also in postnatal liver. At the same time, apparent ACAT activity levels decreased in foetal and regenerating rat livers when compared with respective control tissues. These results suggest that serum cholesterol esters, circulating LCAT and cellular ACAT levels are modulated during cell proliferation.


We describe a novel, testable theory of autoimmunity, outline novel predictions made by the theory, and illustrate its application to unravelling the possible causes of idiopathic thrombocytopenia purpura (ITP). Pairs of stereochemically complementary antigens induce complementary immune responses (antibody or T-cell) that create loss of regulation and civil war within the immune system itself. Antibodies attack antibodies creating circulating immune complexes; T-cells attack T-cells creating perivascular cuffing. This immunological civil war abrogates the self-nonself distinction. If at least one of the complementary antigens mimics a self antigen, then this unregulated immune response will target host tissues as well. Data demonstrating that complementary antigens are found in some animal models of autoimmunity and may be present in various human diseases, especially ITP, are reviewed. Specific mechanisms for preventing autoimmunity or suppressing existing autoimmunity are derived from the theory, and critical tests proposed. Finally, we argue that Koch's postulates are inadequate for establishing disease causation for multiple-antigen diseases and discuss the possibility that current research has failed to elucidate the causes of human autoimmune diseases because we are using the wrong criteria.


High-throughput custom designed genotyping arrays are a valuable resource for biologically focused research studies and increasingly for validation of variation predicted by next-generation sequencing (NGS) technologies. We investigate the Illumina GoldenGate chemistry using custom designed VeraCode and sentrix array matrix (SAM) assays for each of these applications, respectively. We highlight applications for interpretation of Illumina generated genotype cluster plots to maximise data inclusion and reduce genotyping errors.We illustrate the dramatic effect of outliers in genotype calling and data interpretation, as well as suggest simple means to avoid genotyping errors. Furthermore we present this platform as a successful method for two-cluster rare or non-autosomal variant calling. The success of high-throughput technologies to accurately call rare variants will become an essential feature for future association studies. Finally, we highlight additional advantages of the Illumina GoldenGate chemistry in generating unusually segregated cluster plots that identify potential NGS generated sequencing error resulting from minimal coverage.We demonstrate the importance of visually inspecting genotype cluster plots generated by the Illumina software and issue warnings regarding commonly accepted quality control parameters. In addition to suggesting applications to minimise data exclusion, we propose that the Illumina cluster plots may be helpful in identifying potential in-put sequence errors, particularly important for studies to validate NGS generated variation. The generation of next-generation sequencing (NGS) quencies ,2. For cThe Illumina platform  has proven reliable and efficient for a number of high-throughput genotyping applications using DNA extracted from several sources, -9. VeraCIn this study, we assess genotype information generated on custom-designed Illumina 96-SNP VeraCode and BeadArray Sentrix Array Matrix (SAM) assays, for optimal generation of accurate genotyping and NGS generated variant validation. Application of strict analysis parameters may lead to valuable genetic information being lost. Alternatively, limited QC may result in the addition of incorrect genotype calls confounding data analysis. The SNP assays described in this study have the potential of being disregarded or falsely included during analysis based on the current QC selection criteria. We discuss the importance of visually inspecting each cluster plot, particularly for custom designed genotype arrays, and suggest strategies for interpreting data in plots that would normally be discarded during the QC process.Sarcophilus harrisii). DNA was genotyped as part of a validation of sequence variation determined from minimal coverage (0.3× versus 0.5×) of NGS de novo sequencing data of two animals using the Roche/454 Titanium chemistry (unpublished data). Genotype analysis was performed using a custom 96-plex SAM array for 96 samples. For both studies, the GoldenGate genotyping procedure was performed as outlined by Illumina [Two sample data sets were used to assess and interpret Illumina-generated genotype cluster plots. The first was part of a gene/pathway targeted human prostate cancer association study. In brief, DNA was extracted from dried blood spots (Guthrie card) and genotyped using a custom 96-plex VeraCode SNP array for 768 samples (unpublished data). In the second study, DNA was extracted from ear clippings of Tasmanian devils , the eight VeraCode runs were assessed individually. The genotype cluster plots generated by each VeraCode and SAM assays were visually inspected for quality of calls. Plots that appeared to be "unusually" clustered ) were further investigated by selecting samples to assess by direct Sanger sequencing for genotype confirmation. Samples were sequenced using Big Dye Terminator v3.1  chemistry according to manufacturer's guidelines and sequenced on an AB 3730 genetic analyzer (AB).Using the VeraCode technology we observed several consistent examples of SNP cluster plots that appeared to be incorrectly genotyped due to a combination of a short θ and the presence of outlier DNA samples. In the two examples discussed in this study Figure , the previce versa for the X-linked variants , genotyping technologies must accurately perform two-cluster calling. In this study successful two-cluster calling was observed for both rare autosomal , direct Sanger sequencing revealed a single nucleotide error in the NGS generated data surrounding the putative SNP. In the examples highlighted in this study  Large-scale custom-designed genotyping studies have been made feasible by the development of high-throughput technologies and improvements of automated genotype calling software. In this study we demonstrate how simple applications for interpreting automatically generated Illumina cluster plots can be used to avoid spurious genetic associations as well as optimising data inclusion and interpretation, particularly for custom designed target arrays.Variation in quality and quantity of DNA input, is largely unavoidable in large-scale studies. In order to limit the effect potential outlier DNA samples may have on genotype calls, it is important to visually inspect the genotype plots and exclude outliers as demonstrated in this study Figure . In addiInvestigating the disease-causing potential of rare variants , can prde novo generated NGS data, we observed several examples of individual clusters being partially segregated within the one genotype group  which lead to the identification of NGS-generated sequencing error. We therefore suggest that visual inspection and closer investigation of unusually clustered scatter plots, may provide information that exceeds the initial goal of SNP validation and is a complimentary tool for NGS data validation.A growing number of laboratories are employing the use of NGS technology to sequence as yet unclassified genomes, as well as resequencing of human and human disease-associated genomes. From the predicted sequence variants generated by these projects, custom-designed arrays are being employed for validation and frequency determination. Although the optimal coverage required to distinguish true sequence variation is highly debatable, and dependent on the type of NGS platform used, high-throughput genotyping platforms allow for rapid, cost-effective validation at even minimal coverage. The SAM array discussed in this study was developed from long-read NGS information generated from minimal coverage of two samples of an as yet unsequenced species (unpublished data). In both the VeraCode assay, custom designed to genotype 96 confirmed human variants, and the SAM array, designed to verify newly identified variants from p Figure  and 4. Ap Figure , the unuWe demonstrate in this study, applications to optimise and improve the efficiency of data analysis generated using the Illumina GoldenGate chemistry using logical interpretation of both rare and common genotyping data. We also present this platform as a successful tool for NGS variant validation, which is applicable to even limited sequence coverage.The authors declare that they have no competing interests.WM and SCS provided analysis of NGS data, EAT, DCP and SN performed genotyping and Sanger sequencing analysis; EAT, DCP, SN and VMH were involved in data analysis; and EAT and VMH participated in study design and manuscript drafting. All authors read and approved the final manuscript.


Many growth factors are implicated in the pathogenesisof proliferative diabetic retinopathy. Alteration of growthfactors and their receptors in diabetes has been shown inboth experimental and clinical studies. Sustained hyperglycemiaresulting from long-standing diabetes leads to severalbiochemical abnormalities that consequently result inretinal hypoxia. Retinal oxygenation state regulates variousgrowth factors that promote angiogenesis in order to meetthe oxygen demands of the tissue. However, unregulated expressionof these growth factors and induction of complexcascades leading to augmentation of other proangiogenicfactors, which may not be regulated by tissue oxygenation,leads to uncontrolled retinal neovascularization and blindnessin diabetic patients.


Complete mitochondrial genome sequences have become important tools for the study of genome architecture, phylogeny, and molecular evolution. Despite the rapid increase in available mitogenomes, the taxonomic sampling often poorly reflects phylogenetic diversity and is often also biased to represent deeper (family-level) evolutionary relationships.Solenopsis, which represent various evolutionary depths. Overall, ant mitogenomes appear to be typical of hymenopteran mitogenomes, displaying a general A+T-bias. The Solenopsis mitogenomes are slightly more compact than other hymentoperan mitogenomes (~15.5 kb), retaining all protein coding genes, ribosomal, and transfer RNAs. We also present evidence of recombination between the mitogenomes of the two conspecific Solenopsis mitogenomes. Finally, we discuss potential ways to improve the estimation of phylogenies using complete mitochondrial genome sequences.We present the first fully sequenced ant (Hymenoptera: Formicidae) mitochondrial genomes. We sampled four mitogenomes from three species of fire ants, genus The ant mitogenome presents an important addition to the continued efforts in studying hymenopteran mitogenome architecture, evolution, and phylogenetics. We provide further evidence that the sampling across many taxonomic levels (including conspecifics and congeners) is useful and important to gain detailed insights into mitogenome evolution. We also discuss ways that may help improve the use of mitogenomes in phylogenetic analyses by accounting for non-stationary and non-homogeneous evolution among branches. There has been a rapid proliferation of whole mitochondrial genomes (mitogenomes) sequenced in recent years, no doubt driven in part by the increasing speed and decreasing cost of sequencing technologies. Whole mitogenomes are increasingly used in phylogenetic studies -6 and inHowever, the utility of these datasets for these purposes greatly depends on taxon sampling. Currently, 237 insect mitogenomes have been fully sequenced , yet the taxa utilized for these sequencing studies often do not reflect the distribution of species diversity. For example, Hymenoptera is one of the most species-rich insect orders , yet onNasonia  and Drosophila yakuba [GenBank: NC001322]. We subsequently designed additional primers spanning other regions with no or low coverage using sequence data generated for fire ants. All primers developed and used for this study are presented in Additional file Complete mitochondrial genomes were generated for four individuals from three closely related  Brazil) . We sequ Brazil)  as well Several lines of evidence suggest that our sequences generated specifically represent mitochondrial genomic DNA rather than nuclear mitochondrial-like sequences (numts), which appear to be common and are generally short and highly fragmented in ants and other Hymenoptera -83: The HQ215537, HQ215538, HQ215539, HQ215540].All PCR amplicons were sequenced in both directions and each strand was assembled into single contigs with overlapping ends, indicating that our mitogenome sequences contained no gaps. Leading and lagging strand for each mitogenome were then aligned and manually checked for indels or ambiguous base calls. Mitogenomes were deposited in the NCBI GenBank database , 3 beetles , and 3 moths ). jModeltest  and sub-tree pruning and regrafting [SPR]) on five random starting trees. In addition to running one hundred bootstrap replicates to estimate levels of branch support, we also implemented the SH-like aLRT, which assesses the likelihood gain of the presence of that branch [Maximum likelihood analyses were implemented on the PhyML 3.0 webserver http://wwt branch . To accot branch  using deThe evolution of tRNA-N was studied using phylogenetic analyses as suggested by Saks et al.  and DowtJC and DDS carried out all aspects of the molecular lab work. DG and DDS performed all analyses of the data and wrote the manuscript. All authors read and approved the final manuscript.Primers used for PCR amplification and sequencing of fire ant genomes. Primers names and sequences used in this study. J and N within primer names refer to heavy and light strands, respectively, and indicate the orientation of the primers. Primer names beginning with "Gem" were designed specifically to amplify the mitogenome of S. geminata.Click here for file


Lindbergh.Pacemakers have evolved over a period of time trying to mimic the normal response rates, conduction and activation characteristics, though are still far from what nature has bestowed upon us. Better understanding of cardiac physiology and hemodynamics has led to current available pacing technology and we do recognize now that to achieve physiological pacing we should have an appropriate heart rate response, ventriculo-ventricular (VV) synchronization and atrio-ventricular (AV) synchronization.Patients receiving rate responsive pacemakers for sinus node dysfunction, in spite of using various sensors and rate response algorithms -5, stillRight ventricular (RV) pacing represents a non-physiological activation of the heart causing wide QRS (left bundle branch block) with electrical and mechanical VV dyssynchrony . Higher Optimal AV interval at rest ranges from 100 to 150 milliseconds. In normal individuals the AV interval shortens with increased heart rate during exercise in a predictable and linear fashion. Most pacemakers have a programmable shortening of AV delay at higher rates, the hemodynamic benefits of which have not yet been shown . Device An electrocardiogram based method to determine optimal AV interval is described by Sorajja et al  in this Based on echocardiographic parameters and natriuretic peptide levels, AV delay optimization is found to be beneficial in patients with normal LV function in short term small studies -25. TherAdopt the pace of nature:  her secret is patience  - Ralph Waldo EmersonLook deep into nature, and then you will understand everything better  - Albert Einstein


Tibialis posterior has a vital role during gait as the primary dynamic stabiliser of the medial longitudinal arch; however, the muscle and tendon are prone to dysfunction with several conditions. We present an overview of tibialis posterior muscle and tendon anatomy with images from cadaveric work on fresh frozen limbs and a review of current evidence that define normal and abnormal tibialis posterior muscle activation during gait. A video is available that demonstrates ultrasound guided intra-muscular insertion techniques for tibialis posterior electromyography.Current electromyography literature indicates tibialis posterior intensity and timing during walking is variable in healthy adults and has a disease-specific activation profile among different pathologies. Flat-arched foot posture and tibialis posterior tendon dysfunction are associated with greater tibialis posterior muscle activity during stance phase, compared to normal or healthy participants, respectively. Cerebral palsy is associated with four potentially abnormal profiles during the entire gait cycle; however it is unclear how these profiles are defined as these studies lack control groups that characterise electromyographic activity from developmentally normal children. Intervention studies show antipronation taping to significantly decrease tibialis posterior muscle activation during walking compared to barefoot, although this research is based on only four participants. However, other interventions such as foot orthoses and footwear do not appear to systematically effect muscle activation during walking or running, respectively. This review highlights deficits in current evidence and provides suggestions for the future research agenda. The tibialis posterior (TP) muscle has a vital role during gait; via multiple insertion points into the tarsal bones it acts as the primary dynamic stabiliser of the rearfoot and medial longitudinal arch (MLA) ,2. The sThe TP muscle is contained within the deep posterior compartment of the lower limb, arising from the adjacent posterior surfaces of the tibia, fibula and interosseus membrane Figure . The tenThe TP tendon has multiple insertions within the foot, dividing into three main components: (i) anterior; (ii) middle; and (iii) posterior -23. The The most common modality used to quantify TP muscle activation is via EMG recorded with intramuscular electrodes. The advantage of using EMG over other modalities (such as MRI and ultrasound) is the ability to investigate muscle activation simultaneously with dynamic weightbearing tasks such as walking. However, due to the deep location of the muscle within the posterior compartment of the leg, surface electrodes cannot record TP EMG activity without signal cross-talk from various superficial muscles Figure . Therefo(i) the posterior-medial; and (ii) the anterior insertion. A video demonstration of both approaches can be viewed via downloadable supplements  with the use of ultrasound guidance. All five electrodes were correctly located in the muscle belly of TP; figure Experience gained (GSM) in performing more than 150 intramuscular EMG electrode insertions into TP has led to some important practical insights. Participants usually describe low to mild discomfort during the insertion procedure with approximately 1 in 20 describing severe pain, although this has not been quantified using a validated pain scale. When participants experience severe pain, the wires are removed and a second attempt at relocating new wire electrodes is undertaken; rarely is a third attempt required. During walking, participants usually describe 'mild' pain for the first couple of minutes, which frequently subsides to 'no' or 'low' pain after this period. Mild calf pain is often experienced for 24 hours following the insertion procedure. There were no reported cases of serious complications such as infection. The use of wire electrodes is generally a safe and effective method of investigating tibialis posterior EMG during walking.Current literature has characterised TP EMG during gait among normal and pathological populations and with various interventions including antipronation taping, foot orthoses and athletic footwear. Figure Normative EMG for TP during walking is based on studies with typically small sample sizes (ranging from 5 to 12) and with participants' age ranging from 18 to 76 years ,12,27-29TP EMG activity during running was characterised by Reber and colleagues  when theOverall, the availability of normative EMG for TP during walking is based on relatively small sample sizes and is limited to only adult and older adult participants. Despite the absence of normative data, other studies have investigated TP EMG activation with pathological conditions including rheumatological and neurogenic diseases. With the high variability seen in healthy people, it is difficult to conclude whether the findings from studies investigating abnormal muscle activity are meaningful.Tibialis posterior tendon dysfunction (TPTD) has been reported as the most common cause of adult acquired flatfoot ,22,31,32It has been suggested that certain rheumatological conditions may predispose to TPTD including rheumatoid arthritis (RA) ,39 and sDespite the uncertainty regarding the pathogenesis of pes plano valgus, gait analysis has been shown to improve our understanding of this condition in RA ,45. Yet TP muscle dysfunction is likely to occur with many neurogenic conditions, however little is known about how many of these conditions affect TP muscle activity during walking. Cerebral palsy is one neurogenic condition where TP muscle activation has been investigated as part of several laboratory-based clinical assessments -53. CereIntramuscular electrodes have been utilised to assess TP muscle activation among infants, children and young adult patients (age range: 4–24 years) – often as part of a surgical planning procedure Figure  46-50].-50.46-50A further study by Michlitsch and colleagues  involvedWhile there is consensus among these investigations that both TP and tibialis anterior contribute to varus or equinovarus foot deformity with cerebral palsy, one major shortcoming is that none of these investigations have directly compared TP EMG profiles to age matched control groups within the same study. This may account for the different and potentially invalid classifications among these studies of TP dysfunction with cerebral palsy. Further normative EMG from TP is required to inform studies investigating pathological sub-types of TP muscle dysfunction in children with cerebral palsy.Only one study has investigated the effect of foot orthoses on TP activation during walking  despite These findings contrast another investigation on the effect of anti-pronation taping on TP EMG during walking in four young- to middle-aged adults with flat-arched feet . FranettA number of studies investigating TP EMG activation in health and disease have been undertaken with small sample sizes providing preliminary evidence of either abnormal function or response to intervention. Accordingly, further EMG studies, recruiting larger sample sizes and representation from the younger and older populations, are required to investigate both the effect of interventions on TP muscle activity and to establish a reference database. Whilst it has been recognised that TP plays a vital role during gait, further work is required to more fully understand the role of TP in the development of pathology and in disease-specific populations including RA, cerebral palsy and TPTD.In summary, TP EMG remains a specialist investigation undertaken in relatively few centres internationally; however, this technique has multiple applications both in research and in planning interventions and evaluating outcomes. Recent advances in technology, including imaging, represent an opportunity to employ this technique more frequently and advance our understanding in a variety of areas.The authors declare that they have no competing interests.DET and JW conceived the idea for the review, RS and GSM drafted the manuscript and the figures, GSM prepared the video supplement, JW and DET critically revised the manuscript. All authors read and approved the final manuscript.Posterior approach. A video demonstration of the posterior approach of intramuscular electrode insertion. Additional files 1 and 2 can only be viewed using the latest version of QuickTime Player which can be downloaded via the following link: http://www.apple.com/downloads/Click here for fileAnterior approach. A video demonstration of the anterior approach of intramuscular electrode insertion.Click here for file


We report a case of a rapid renal deterioration due to ureteropelvic junction obstruction (UPJO) in an asymptomatic woman with prior normal diuretic renography. This case illustrates “silent” renal obstruction and the inability of diuretic renography in detecting significant renal obstruction. This case may favor close surveillance of any adult patient with potential UPJO, especially those with underlying renal disease or solitary kidney. Ureteropelvic junction obstruction (UPJO) is an uncommon condition in adults. UPJO is most commonly diagnosed during childhood with treatment required for any patient demonstrating renal deterioration, stone formation, infection, or pain. Not all patients with hydronephrosis require intervention, although the natural history of untreated UPJO is better understood in children than adults. We describe a woman who rapidly developed renal deterioration due to UPJO despite lacking symptoms and previous normal evaluation by diuretic renography.A 62-year-old woman was referred for a brief episode of left flank pain and imaging demonstrating fullness of the left renal pelvis . She hadAt the request of her primary care physician, the patient underwent repeat computed tomography (CT) imaging 12 months later despite being asymptomatic. Repeat imaging demonstrated severe left-sided hydronephrosis . Repeat The natural history of UPJO in adulthood is poorly understood but believed to be related to intrinsic narrowing of the upper ureter or extrinsic pressure on the ureter caused by aberrant vessels or fibrous bands. As prenaAlthough there is evidence that not all pediatric patients demonstrating hydronephrosis require intervention, the natural history of untreated UPJO in adults is less well documented. Most studies document preservation or improvement of renal function following treatment for adult UPJO.–5 A smalThe case presented documents rapid renal deterioration from UPJO in an asymptomatic adult patient with prior normal evaluation for presence of obstruction. Although undoubtedly rare, this case illustrates that diuretic renography may not detect significant renal obstruction, and that renal deterioration may occur in an otherwise asymptomatic patient. The sensitivities of current standards for the diagnosis of UPJO, namely diuretic renography and CT scanning, are imperfect. There remains room for improvement in the diagnosis, perhaps at a functional imaging level or at a molecular level, of UPJO.Nonetheless, serial imaging in all asymptomatic patients who have a normal evaluation for the presence of UPJO cannot be recommended, however repeat imaging (ultrasound or CT scanning) may be prudent in patients at risk for renal compromise . Patients and physicians need be aware of silent renal obstruction and the potential for diuretic renography and CT scanning to, at least transiently, miss the diagnosis.


The streptococcal preparation OK432 was studied for its effects on natural killer (NK) activity of peripheral blood lymphocytes (PBL) from normal donors and from ovarian cancer patients, and of tumour-associated lymphocytes (TAL) from peritoneal effusions. OK432 augmented NK activity against the susceptible K562 line and induced killing of the relatively resistant Raji line. Freshly isolated ovarian carcinoma cells were relatively resistant to killing by unstimulated PBL and TAL. OK432 induced significant, though low, levels of cytotoxicity against 51Cr-labelled ovarian carcinoma cells. Augmentation of killing of fresh tumour cells by OK432 was best observed in a 20 h assay and both autologous and allogeneic targets were lysed. PBL were separated on discontinuous Percoll gradients. Unstimulated and OK432-boosted activity were enriched in the lower density fractions where large granular lymphocytes (LGL) and activity against K562 were found. Thus, OK432 augments NK activity of PBL and TAL in human ovarian carcinomas and induces low, but significant, levels of killing of fresh tumour cells. Effector cells involved in killing of fresh ovarian tumours copurify with LGL on discontinuous gradients of Percoll.


A new imaging modality framework, calledelasto-mammography, is proposed to generate the elastograms ofbreast tissues based on conventional X-ray mammography. Thedisplacement information is extracted from mammography projectionsbefore and after breast compression. Incorporating thedisplacement measurement, an elastography reconstruction algorithmis specifically developed to estimate the elastic moduli ofheterogeneous breast tissues. Case studies with numerical breastphantoms are conducted to demonstrate the capability of theproposed elasto-mammography. Effects of noise with measurement,geometric mismatch, and elastic contrast ratio are evaluated inthe numerical simulations. It is shown that the proposedmethodology is stable and robust for characterization of theelastic moduli of breast tissues from the projective displacementmeasurement.


Recent preclinical work investigating the role of progenitor cell therapies for central nervous system (CNS) injuries has shown potential neuroprotection in the setting of traumatic brain injury (TBI), spinal cord injury (SCI), and ischemic stroke. Mechanisms currently under investigation include engraftment and transdifferentiation, modulation of the locoregional inflammatory milieu, and modulation of the systemic immunologic/inflammatory response. While the exact mechanism of action remains controversial, the growing amount of preclinical data demonstrating the potential benefit associated with progenitor cell therapy for neurological injury warrants the development of well-controlled clinical trials to investigate therapeutic safety and efficacy. In this paper, we review the currently active or recently completed clinical trials investigating the safety and potential efficacy of bone marrow-derived progenitor cell therapies for the treatment of TBI, SCI, and ischemic stroke. Our review of the literature shows that while the preliminary clinical trials reviewed in this paper offer novel data supporting the potential efficacy of stem/progenitor cell therapies for CNS injury, a great deal of additional work is needed to ensure the safety, efficacy, and mechanisms of progenitor cell therapy prior to widespread clinical trials. Central nervous system (CNS) insults including ischemic stroke, traumatic brain injury (TBI), and traumatic spinal cord injury (SCI) represent a major burden to the healthcare system worldwide. Approximately 5 million people are burdened by the long-term physical, cognitive, and psychosocial deficits associated with TBI in the United States with 40% of patients reporting unmet needs 1 year post injury . OverallPreliminary research is currently underway to evaluate the potential efficacy of adult tissue progenitor (stem) cell therapies for the treatment of CNS injury. Adult tissue progenitor cells are located in select microenvironments (niches) which protect against overproliferation and depletion as well as regulate progenitor cell involvement in resident tissue repair and regeneration . By defiRecent preclinical work investigating bone marrow derived-progenitor cell therapies for CNS injury has shown potential neuroprotection after TBI , ischemiin vitro with subsequent implantation into rodent cortex after focal ischemic injury. The implanted progenitor cells were found to display neural cell markers and engraft up to 100 days after injury with associated behavioral recovery ..46].Geron Corporation  has started a United States Food and Drug Administration (FDA) approved trial using embryonically derived oligodendroglial precursor cells  to replace myelin-forming cells within injured spinal cords. The cell preparations are injected directly into the spinal cords at the lesion site. The treatment population is restricted to adults with complete thoracic (T3–T9) level SCI. Patients must undergo treatment within 7 to14 days following injury. Preclinical data showing that SCI animals treated with Geron's cell line developed cysts at the level of treatment caused the FDA to put a hold on the clinical trial. Geron and the FDA have reached an agreement to allow the trail to move forward if subsequent preclinical studies provide satisfactory outcome.in vivo cell tracking need to be completed to ensure the safety of potential trials while affording them the best possible chance at success. Additional preclinical work to more clearly delineate the progenitor cell mechanism of action would also aid in the planning of quality-controlled clinical studies. Overall, while the very preliminary clinical trials reviewed in this paper offer novel data supporting the potential efficacy of cell therapeutics for CNS injury, a great deal of additional work is needed to ensure the safety and efficacy of progenitor cell therapy prior to widespread clinical trials.Prior to large, multicenter clinical trials investigating the potential efficacy of progenitor cell therapies for CNS insults, a number of issues need to be addressed. Further research into optimal cell dosing, cell delivery method, and techniques for


Fungal diseases are increasing among patients infected with human immunodeficiency virus (HIV) type 1. Infections due to Candida and Cryptococcus are the most common. Although mucocutaneous candidiasis can be treated with oral antifungal agents, increasing evidence suggests that prolonged use of these drugs results in both clinical and microbiologic resistance. The optimal therapy for cryptococcal meningitis remains unresolved, although initial treatment with amphotericin B, followed by life-long maintenance therapy with fluconazole, appears promising. Most cases of histoplasmosis, coccidioidomycosis, and blastomycosis occur in regions where their causative organisms are endemic, and increasing data suggest that a significant proportion of disease is due to recent infection. Aspergillosis is increasing dramatically as an opportunistic infection in HIV-infected patients, in part because of the increased incidence of neutropenia and corticosteroid use in these patients. Infection due to Penicillium marneffei is a rapidly growing problem among HIV-infected patients living in Southeast Asia. Although the advent of oral azole antifungal drugs has made primary prophylaxis against fungal diseases in HIV-infected patients feasible, many questions remain to be answered before the preventive use of antifungal drugs can be advocated.


A(PDPK FA) was overexpressed in various cancerous tissues relative to normal controls. However, the functional role of overexpressed PDPK FAin cancer remains to be established. In this report, we explore the potential role of PDPK FAin leukaemia cell growth by investigating the effects of partial inhibition of this kinase on the malignant phenotype of human chronic myeloid leukaemia cells (K562). Cloning of PDPK FAcDNA and its recombinant antisense expression vector and PDPK FA-specific antibody were successfully developed. Two stable antisense clones of K562 cells were subcloned which expressed 70% and 45% of PDPK FArespectively, compared with control-transfected clone in both immunoprecipitate activity assay and immunoblot analysis. In sharp contrast, these two antisense clones expressed no significant suppression of any other related PDPK family members, indicating the specificity of these two antisense clones. Moreover, these antisense clones proportionally and potentially exhibited cell growth retardation, poor clonogenic growth in soft agar and loss of serum independence. The results demonstrate that specific antisense suppression of PDPK FAis sufficient to interfere with the growth of K562 cells, indicating that PDPK FAis essential for human chronic myeloid leukaemia cell growth. © 2000 Cancer Research CampaignInitial studies revealed that proline-directed protein kinase F


The importance of the applications of lanthanides, as an excellent diagnosticand prognostic probe in clinical diagnostics, and an anticancer material, is remarkably increasing. Lanthanidecomplexes based X-ray contrast imaging and lanthanide chelates based contrast enhancing agents formagnetic resonance imaging (MRI) are being excessively used in radiological analysis in our body systems.The most important property of the chelating agents, in lanthanide chelate complex, is its ability to alter thebehaviour of lanthanide ion with which it binds in biological systems, and the chelation markedly modifiesthe biodistribution and excretion profile of the lanthanide ions. The chelating agents, especially aminopolycarboxylic acids, being hydrophilic, increase the proportion of their complex excreted from complexedlanthanide ion form biological systems. Lanthanide polyamino carboxylate-chelate complexes are used ascontrast enhancing agents for Magnetic Resonance Imaging. Conjugation of antibodies and other tissuespecific molecules to lanthanide chelates has led to a new type of specific MRI contrast agents and theirconjugated MRI contrast agents with improved relaxivity, functioning in the body similar to drugs. Manyspecific features of contrast agent assisted MRI make it particularly effective for musculoskeletal andcerebrospinal imaging. Lanthanide-chelate contrast agents are effectively used in clinical diagnosticinvestigations involving cerebrospinal diseases and in evaluation of central nervous system. Chelatedlanthanide complexes shift reagent aided


Peripheral-blood monocytes from normal individuals and from patients with malignant melanoma reduce nitroblue tetrazolium (NBT). A quantitative assay for dye reduction was applied to 25 healthy donors and 31 patients with malignant melanoma. NBT reduction expressed as dye reduction per monocyte was significantly impaired in patients with disseminated disease, and they responded poorly to a phagocytic stimulus. Monocytes from patients with micrometastatic disease, however, showed normal resting NBT reduction but, following exposure to a suspension of latex-polystyrene, showed significantly greater NBT reduction than those from normal individuals. Since NBT reduction is an indirect measure of intracellular hexose-monophosphate-shunt activity we conclude that the monocytes from patients with minimal disease are in some way activated.


The STAR-VA training program in Veterans Health Administration Community Living Centers (CLCs) has been helping interdisciplinary care teams understand and manage dementia-related behaviors in the nursing home setting, with promising clinical outcomes. However, sustaining a new care approach in a health care system poses multiple challenges. This presentation will discuss facilitators and barriers to STAR-VA sustainability based on CLC team and nurse leader feedback. Findings are informing development of a new site coaching program and a sustainability toolkit. Feedback to date suggests that critical STAR-VA implementation and sustainability strategies include: regularly scheduled team meetings to discuss behavioral assessment and care plans; ongoing staff training ; communicating care plans across shifts and in the health record; multiple nurse/shift champions; impromptu huddles; acknowledging staff successes; leadership engagement. The coaching program engages teams in setting and tracking site sustainability goals. Lessons learned will be discussed.


Persons with dementia frequently demonstrate behavior symptoms of dementia (BSD), associated with poorer outcomes. A measure of BSD was created for routine use in VA Community Living Centers (CLCs). Reliability and validity of Minimum Data Set (MDS 3.0) behavior items was established using exploratory factor analysis and a multitrait, multimethod correlation matrix. 385 CLC residents with BSD were assessed using validated measures of BSD, depression, and anxiety, and team ratings of the frequency and severity of target behaviors identified for intervention. Factor analysis on MDS items closest to baseline resulted in two stable factors. MDS behavior factors related to validated clinical measures in predicted ways at baseline and post-intervention. MDS distress behavior factor sensitivity to change was evaluated by using change score correlations with validated clinical measures. The MDS distress behavior factor can be used routinely, evaluate the impact of intervention effectiveness, and provide quality improvement feedback.


Primary immunodeficiencies (PID) are a heterogeneous group of disorders with a variable clinical spectrum of manifestations. The central nervous system may be involved in PID with symptoms which may present initially or develop at later stages. The purpose of this study was to review the neurological manifestations of different PID syndromes.We focused on 104 selected studies on PID with certain neurological abnormalities which may accompany these disorders or may later signify a PID in their course.Diverse neurological deficits accompanying certain PIDs may be mild or they may greatly influence the course of the disease with major impacts on the quality of life of these patients.Early recognition and treatment is important to prevent or reduce future irreversible neurological sequelae. Therefore physicians should be aware of the neurological features accompanying PID. Primary immunodeficiencies (PID) are a heterogeneous group of 354 distinct disorders with 344 different gene defects  with a vPIDs have been classified practically according to the affected immune function to the following groups: immunodeficiencies affecting cellular and humoral immunity, combined immunodeficiencies with associated or syndromic features, antibody deficiencies, diseases of immune dysregulation, congenital defects of phagocytes, defects in intrinsic and innate immunity, autoinflammatory disorders, complement deficiencies and phenocopies of PIDs . The cenNeurological manifestations will be discussed in detail according to the specific classification of primary immunodeficiencies to assist the treating physicians’ timely diagnosis and prompt treatment in order to avoid irreversible neurologic sequelae. The categorization of the inborn errors of immunity is based on the International Union of Immunological Societies: 2017 Primary Immunodeficiency Diseases Committee Report on Inborn Errors of Immunity ADA is a ubiquitous enzyme in purine salvage pathway which is also expressed in both the peripheral and central nervous systems . ADA defDNA Ligase IV deficiencyThis autosomal recessive form of SCID is caused by an impairment of the DNA damage repair process with a pronounced radiosensitivity . DNA douCernunnos deficiencyThis is another rare autosomal recessive form of SCID with severe T and B lymphopenia and dysgammaglobulinemia in addition to radiosensitivity caused by mutations in the CERNUNNOS or XRCC4-like factor (XLF). Microcephaly and developmental delay are the prominent neurological features . CID with associated or syndromic featuresAtaxia-TelengiectasiaThis autosomal recessive complex disorder with substantial severity in affected individuals  is charaThere are also Ataxia Telengiectasia-Like Disorders (ATLD) which are clinically similar to ataxia telengiectasia although without telangiectasia and a slower progression of neurologic manifestations. These patients remain ambulatory until their third decades of lives , 20.Nijmegan Breakage SyndromeThis is a rare autosomal recessive, multisystemic disease of chromosomal instability presenting at birth with microcephaly but no additional neurologic manifestation . The mutImmunodeficiency, Centromeric Region Instability and Facial Anomalies SyndromeThis genetic disease is due to a mutation in DNA methylteransferase gene  which caRiddle SyndromeThis syndrome is characterized by a defect in DNA damage response causing immunodeficiency and increased radiosensitivity accompanied by neurologic symptoms and growth delay due to an increased apoptosis and reduced proliferative capacity during brain development presenting as mild motor control and learning difficulties . DiGeorge SyndromeIn addition to the characterizing triad of cardiac defects, hypocalcemia and cellular immune deficiency, there may be neurological manifestations which present with developmental delay in motor, language and speech areas, neuropsychiatric problems and epilepsy . Most paHyper-immunoglobulin E syndromeThese syndromes are characterized by recurrent skin and pulmonary infections in the presence of elevated IgE concentrations and usually eosinophilia. The hyper-IgE syndromes have four distinct genetic causes including mutations in signal transducer and activator of transcription 3 (STAT3), dedicator of cytokinesis 8 (DOCK8), tyrosine kinase2 (TYK2) and phosphoglucomutase 3 (PGM3) , 34. VasSchimke Immuno-osseous DysplasiaThis autosomal recessive disorder is caused by mutation in the SMARCAL1 gene which encodes a chromatin remodeling protein. Defective cellular immunity causes recurrent bacterial, fungal and viral infections which in association with short stature and progressive renal insufficiency characterizes the disorder . NeuroloHoyeraal-Hreidarsson syndromeThis is an X-linked recessive disorder of telomere dysfunction which represents a severe form of dyskeratosis congenita characterized by a classic triad of reticular skin pigmentation, dysplastic nails and oral leukoplakia . PancytoPurine Nucleoside Phosphorylase DeficiencyThis rare autosomal recessive metabolic disorder, similar to ADA deficiency, results from impaired purine salvage pathway which causes combined immunodeficiency in association with prominent neurological abnormalities . “MutatiHepatic Veno-Occlusive Disease with ImmunodeficiencyThis is caused by homozygous mutation in the autosomal SP110 gene. The onset of clinical features usually occurs before the age of six months with hepatic sinusoidal obstruction. Serum Ig levels are low. T cell subsets and B-cell numbers are reduced. Neurological abnormalities include leukodystrophy and extensive cerebral necrosis on postmortem examination are found in one third of the patients .VICI Syndrome This rare autosomal recessive disorder is caused by mutation in the EPG5 gene which results in defective autophagy . ClinicaPredominantly Antibody deficiencyCommon variable immunodeficiencies and agammaglobulinemia  are the most frequent forms of presentations of antibody deficiency. Symptoms mainly consist of respiratory or gastrointestinal involvement. However, neurologic manifestations may also occur. Common Variable ImmunodeficiencyNeurologic manifestations are a complex of diverse and rare complications of CVID including brain or spinal cord infections (meningitis as the most common neurologic manifestation), autoimmune involvements  , transveAgammaglobulinemiaIn patients with agammaglobulinemia symptoms mainly consist of recurrent respiratory or gastrointestinal infections. Serum immunoglobulins are severely decreased and usually undetectable . CNS comDisorders of Immune RegulationChediac Higashi SyndromePatients with this autosomal recessive lethal disorder present with recurrent respiratory and bacterial skin infections due to severe immunodeficiency, partial oculocutaneous albinism, bleeding tendency and progressive neurological features . This syHermansky-Pudlak syndrome type 2Among Hermansky Pudlak syndromes, this type is characterized by an immunodeficiency due to neutropenia and T lymphocyte dysfunction which differentiates it from other types of this syndrome . These pFamilial Hemophagocytic LymphohistiocytosisFHL comprises a group of autosomal recessive life-threatening disorders characterized by uncontrolled proliferation of activated lymphocytes and macrophages associated with massive secretion of cytokines infiltrating into organs including lymph nodes, bone marrow, liver, spleen and central nervous system , 71. CNSCongenital defects of phagocytic number or functionChronic Granulomatous DiseaseChronic granulomatous disease is the most common inherited disorder of phagocytic cells. It results from an inability of phagocytes to produce bactericidal superoxide anions due to defects of the NADPH oxidase system and it leads to the defect in killing of a specific spectrum of bacteria and fungi resulting in concomitant hyper-inflammation and tissue granuloma formation . NeuroloSevere Congenital NeutropeniaThis term is used for a heterogeneous group of primary immunodeficiencies including sporadic autosomal recessive, autosomal dominant and x-linked types  of whichSchwachman-Diamond SyndromeThis is a multisystem disorder characterized by bone marrow failure, exocrine pancreatic insufficiency and metaphyseal chondrodysplasia. These features are attributed to mutation in SBDS gene . HoweverLeukocyte Adhesion Deficiency type 2LAD syndromes consist of rare autosomal recessive disorders among which only LAD type 2 presents with neurologic abnormalities. These patients are characterized by recurrent bacterial infections without pus formation in the first years of life and severe mental retardation later . Other nβ-Actin deficiencyThis autosomal dominant immunodeficiency disorder caused by a non-lethal mutation, characterized by defects in neutrophil migration, mental retardation, short stature, thrombocytopenia and photosensitivity . The halAutoinflammatory DisordersMevalonate Kinase Deficiency (MKD)This is a term used for a wide clinical spectrum of autosomal recessive disorders due to a congenital error in cholesterol biosynthesis, with hyperimmunoglobulinemia D syndrome as the benign variant of the disorder and Mevalonic aciduria as the more severe form of MKD . MevalonNeonatal Onset Multisystem Inflammatory DisorderThe disease results from mutations in the CIAS-1 gene which encodes cryopyrin . CryopyrBlau syndrometh nerve palsy, cerebral vasculitis and cerebral infarction (This rare autosomal dominant syndrome clinically resembles early onset sarcoidosis characterized by granulomatous inflammation mostly involving joints, eyes and skin -97 and ifarction , 101.Aicardi-Goutieres syndromeThis is a genetically determined encephalopathy caused by mutations in TREX1 genes encoding RNAs involved in removing RNA, leading to the accumulation of endogenous RNA, triggering Toll-like receptor-dependent interferon-α production in the brain with the resultant activation of neurotoxic lymphocytes and immune system in addition to the inhibition of angiogenesis , 103. NeIn conclusion, neurological manifestations of primary immunodeficiencies are common with diverse pathologic mechanisms. Neurological deficits may be mild or they may greatly influence the course of the disease with major impacts on the quality of life of the patients. Some of these complications may even cause death. Neurological manifestations may have an early onset, beginning at birth or they may appear later which in both situations may lead to misdiagnosis and delayed therapy. Therefore a high suspicion of an underlying primary immunodeficiency is important in any type of neurological problems occurring with recurrent infections. All three authors were involved in data collection, and writing the article.The authors declare that there is no Conflict of Interest.


Previous studies highlighted the destructive effects of neurotoxin  on integrity of nigral dopamine system, accompanying series of changes in motor regulating network relate to striatal pathways. The fact that L-Dopa or deep brain stimulation (DBS) improves motor gaits in Parkinson's disease (PD) further strengthened the potential importance of subcortical circuits in motor ability control. A recent study highlighted the contribution of cortical inhibition in the process, and therefore might imply the cortical targeting strategy in motor disorder management , which might contribute to the decreased ability of motor learning. This rapid turnover correlates to enhanced apical dendrite Ca2+ spike generation, which is dependent on local NMDA receptors, the major source of calcium permeable glutamate receptors.Somatostatin (SST) interneuron provides important sources to dendritic inhibition, while Parvalbumin (PV) preferably targets somatic region of cortical pyramidal neurons . Indeed, several studies reported aberrant neuronal activity at cortex and GABA deficits in PD patients or animal models of PD. In addition, L-dopa administration enhanced GABA efflux at primary motor cortex, which might be relate to partly the motor improvement, and sometimes the dyskinesia as well . TMS pulses evoke cortical oscillation changes and dis-inhibit cortical dendrites by modulation cortical inhibition (Murphy et al., Taken together, many lines of evidences co-suggest the importance of motor cortical inhibition in motor learning ability, and potentially other aspects of motor functioning. Targeting cortical inhibition with drugs, exercise, and TMS stimulation are feasible approaches for motor rehabilitation.All authors designed the study together and wrote the manuscript together.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.


The goal of cancer eradication has been overshadowed despite the continuous improvement in research and generation of novel cancer therapeutic drugs. One of the undeniable existing problems is drug resistance due to which the paradigm of killing all cancer cells is ineffective. Tumor microenvironment plays a crucial role in inducing drug resistance besides cancer development and progression. Recently, many efforts have been devoted to understand the role of tumor microenvironment in cancer drug resistance as it provides the shelter, nutrition, and paracrine niche for cancer cells. Cancer-associated fibroblasts (CAFs), one major component of tumor microenvironment, reside in symbiotic relationship with cancer cells, supporting them to survive from cancer drugs. The present review summarizes the recent understandings in the role of CAFs in drug resistance in various tumors. Acknowledging the fact that drug resistance depends not only upon cancer cells but also upon the microenvironment niche could guide us to formulate novel cancer drugs and provide the optimal cancer treatment. Various studies have already identified that the nature of tumor does depend not only upon the malignant cancerous cells themselves but also to their microenvironment components Kalluri . The conCAFs constitute major proportion of non-neoplastic stromal compartment in various human tumors. Various researches have suggested that they are capable of modulating tumor cells by forming the communication network with cancer cells or with other elements and hence susceptible to cancer drug resistance  inhibitor in genetically engineered pancreatic ductal adenocarcinoma (PDAC) and chemically induced bladder cancer mice model showed reduction in stromal contents consequently promoting tumor growth and malignancy, which indicated stromal cells function as tumor suppression  and myofibroblast (myCAFs), where they observed that myCAFs lack tumor-promoting chemokines and cytokines , which is regulated by NF-κβ after DNA damage, activates the canonical Wnt program in tumor cells and induces cytotoxic chemotherapy resistance in prostate cancer  breast cancers. A research performed in ER+ MCF-7 cells showed that CAF was the source of tamoxifen and fulvestrant resistance and also protected cancer cells from doxorubicin and PARP inhibitor  was decreased in breast CAFs and promoted cancer invasion by overexpression of thrombospondin type 1  , poor prognostic marker in pancreatic cancer cells, did not inhibit tumor growth and they verified two subtypes: saa3-competent and saa3-null CAFs, which showed pro-tumorigenic and anti-tumorigenic effect, respectively  was blocked by CXCL12 receptor inhibitor  unable to identify its specific origin  (Fujiwara et al. One previous study showed that despite the heterogeneity of CAFs, they are genetically more stable than cancer cells. Therefore, targeting CAFs in cancer treatment would have smaller possibilities to develop drug resistance Kerbel ]. RecentIn conclusions, CAFs, the building block of microenvironment, have a crucial role in modulating cancer drug sensitivity and understanding in detail could minimize the existing challenge of drug resistance and direct the future innovation of cancer drug by avoiding or overcoming the drug resistance.