Dirofilariasis mouse models for heartworm preclinical research.

Introduction: Dirofilariasis, including heartworm disease, is a major emergent veterinary parasitic infection and a human zoonosis. Currently, experimental infections of cats and dogs are used in veterinary heartworm preclinical drug research. Methods: As a refined alternative in vivo heartworm preventative drug screen, we assessed lymphopenic mouse strains with ablation of the interleukin-2/7 common gamma chain (γc) as susceptible to the larval development phase of Dirofilaria immitis. Results: Non-obese diabetic (NOD) severe combined immunodeficiency (SCID)γc-/- (NSG and NXG) and recombination-activating gene (RAG)2-/-γc-/- mouse strains yielded viable D. immitis larvae at 2-4 weeks post-infection, including the use of different batches of D. immitis infectious larvae, different D. immitis isolates, and at different laboratories. Mice did not display any clinical signs associated with infection for up to 4 weeks. Developing larvae were found in subcutaneous and muscle fascia tissues, which is the natural site of this stage of heartworm in dogs. Compared with in vitro-propagated larvae at day 14, in vivo-derived larvae had completed the L4 molt, were significantly larger, and contained expanded Wolbachia endobacteria titres. We established an ex vivo L4 paralytic screening system whereby assays with moxidectin or levamisole highlighted discrepancies in relative drug sensitivities in comparison with in vitro-reared L4 D. immitis. We demonstrated effective depletion of Wolbachia by 70%-90% in D. immitis L4 following 2- to 7-day oral in vivo exposures of NSG- or NXG-infected mice with doxycycline or the rapid-acting investigational drug, AWZ1066S. We validated NSG and NXG D. immitis mouse models as a filaricide screen by in vivo treatments with single injections of moxidectin, which mediated a 60%-88% reduction in L4 larvae at 14-28 days. Discussion: Future adoption of these mouse models will benefit end-user laboratories conducting research and development of novel heartworm preventatives via increased access, rapid turnaround, and reduced costs and may simultaneously decrease the need for experimental cat or dog use.

Gastric teratoma presenting as melena in the newborn: A case report.

Melena is reported in 1 of 350 to 400 new-borns. Significant upper gastrointestinal bleeding in a neonate with an antenatally diagnosed abdominal mass has not been reported before. This case highlights an unusual presentation of a gastric teratoma and proposes a probable embryological explanation for the site of occurrence.

Evaluating the effectiveness of a 'Tobacco Monitor' App in reporting violations of tobacco policy in the community.

It is of interest to evaluate the effectiveness of the "Tobacco Monitor" app in reporting violations of tobacco policy in the community. Hence, a study was conducted amongst the first and second-year undergraduate students of health science colleges of a University. Students were asked to register complaints related to tobacco violations on the tobacco monitor app. Registered complaints were verified by the National Forum for Tobacco Eradication (NFTE) and descriptive statistics were used in reporting the results. A total of 208 complaints on tobacco violation were registered through the Tobacco Monitor app, 163 valid complaints were identified and 45 reports were found invalid. 163 verified valid complaints by NFTE were transferred to the Non-Communicable Diseases (NCD) Cell, Maharashtra, India. It should be noted that anti-tobacco laws and national policies help to curb the menace of the tobacco epidemic to an extent. However, robust reporting and sustainable enforcement measures are required in implementing tobacco legislation effectively. We also report that youth are comfortable in using the Tobacco Monitor app for reporting violations on tobacco.

Neuropsychiatric manifestations in an adolescent male with Rowell syndrome.

A 15-year-old boy presented with fever, skin, and oral lesions for 4 weeks. The cutaneous lesions were suggestive of subacute cutaneous lupus erythematosus and erythema multiforme. His clinical, histopathological, and immunological features were indicative of Rowell syndrome and he satisfied the diagnostic criteria of Rowell syndrome proposed by Zeitouni et al. He subsequently developed neurological manifestations and was diagnosed to have neuropsychiatric systemic lupus erythematosus. We report this case for the unusual occurrence of a rare entity like Rowell syndrome in an adolescent male with co-existence of neuropsychiatric systemic lupus erythematosus.

The applicability of the Demirjian, Willems and Chaillet standards to age estimation of 5-15 year old Indian children.

BACKGROUND: Demirjian's method of age estimation has been reported to overestimate age and Willems' method to give consistently more accurate results. Not enough, however, is known about the applicability of Chaillet's standards. AIM: The present study aimed to compare the accuracy of Demirjian's, Willems' and Chaillet's standards in age estimation of 5 to 15 year-old Indian children. DESIGN: In this cross-sectional observational study, three methods were compared for accuracy in estimating the age of 1200 Indian children aged 5-15 years. RESULTS: Demirjian's method overestimated age by +0.24 ± 0.80 years, +0.11 ± 0.81years and +0.19 ± 0.80 years in boys, girls and the total sample, respectively. With Willems' method, overestimations of +0.09 ± 0.80 years, +0.08 ± 0.80 years and +0.09 ± 0.80 years were obtained in boys, girls and the total sample, respectively. Chaillet's method underestimated age by -0.12 ± 0.69 years, -0.45 ± 0.88 years and -0.25 ± 0.83 years in boys, girls and the total sample, respectively. Statistically significant differences were observed between dental and chronological ages with all methods (p < 0.001). Significant sex-based differences were observed only with Demirjian's and Chaillet's methods (p < 0.05). CONCLUSION: Willems' method was the most accurate in age estimation, followed by Demirjian's and Chaillet's methods. While Demirjian's method was more accurate than Chaillet's in females, Chaillet's method better predicted the age of males.

The signaling axis atypical protein kinase C λ/ι-Satb2 mediates leukemic transformation of B-cell progenitors.

Epigenetically regulated transcriptional plasticity has been proposed as a mechanism of differentiation arrest and resistance to therapy. BCR-ABL leukemias result from leukemic stem cell/progenitor transformation and represent an opportunity to identify epigenetic progress contributing to lineage leukemogenesis. Primary human and murine BCR-ABL+ leukemic progenitors have increased activation of Cdc42 and the downstream atypical protein kinase C (aPKC). While the isoform aPKCζ behaves as a leukemic suppressor, aPKCλ/ι is critically required for oncogenic progenitor proliferation, survival, and B-cell differentiation arrest, but not for normal B-cell lineage differentiation. In vitro and in vivo B-cell transformation by BCR-ABL requires the downregulation of key genes in the B-cell differentiation program through an aPKC λ/ι-Erk dependent Etv5/Satb2 chromatin repressive signaling complex. Genetic or pharmacological targeting of aPKC impairs human oncogenic addicted leukemias. Therefore, the aPKCλ/ι-SATB2 signaling cascade is required for leukemic BCR-ABL+ B-cell progenitor transformation and is amenable to non-tyrosine kinase inhibition.

Neighbourhood influence on the fourth dose of diphtheria-tetanus-pertussis vaccination.

OBJECTIVES: Using 542,159 vaccination records from children born between April 1, 2007, and March 31, 2012, in the Michigan Care Improvement Registry and data from the American Community Survey, we determine if neighbourhood-level characteristics at the Census tract level and block level are associated with low uptake of the fourth dose of diphtheria-tetanus-acellular pertussis vaccine (DTaP4). STUDY DESIGN: This study was a cross-sectional study. METHODS: We used exploratory factor analysis to determine important socio-economic factors at the Census block level and tract level. We then used generalised estimating equations to test the relationship between block- and tract-level socio-economic factors and DTaP4 uptake. RESULTS: DTaP4 coverage was 88.6% (95% confidence interval [CI]: 88.4%-88.7%) in Michigan. At the Census tract level, two factors surfaced as important for DTaP4 vaccination: 'affluence' (Cronbach's alpha = 0.88) and 'socio-economic disadvantage' (Cronbach's alpha = 0.89). At the Census block level, one factor was important: 'affluence' (Cronbach's alpha = 0.90). Affluence may relate to knowledge about medical exemptions and antivaccination sentiment, while socio-economic disadvantage may indicate limited access to healthcare resources. Children in high-affluence tracts had 1.08% lower vaccination coverage (95% CI: -1.62% to -0.55%) than children in low affluence tracts. Children in low socio-economic disadvantage tracts had 2.92% higher coverage than children in high socio-economic disadvantage tracts (95% CI: 2.58%-3.26%). CONCLUSIONS: This study articulates the need to further understand the contribution of neighbourhood-level characteristics, from both affluent and socioeconomically disadvantaged areas to low vaccination rates. Developing a better understanding of these social environmental factors will help determine useful community-level interventions to improve vaccination rates and reduce disease burden.

The applicability of the original and revised Demirjian standards to age estimations of 5-15 year old Indian children.

BACKGROUND: The Demirjian method has been the most widely tested method for the estimation of the dental age of children and adolescents. However, only three studies have compared Demirjian's original and revised seven-tooth methods, four-tooth method and alternate four-tooth method, none of them conducted on an Indian population. AIM: The present study aimed to compare the applicability of the original and revised seven-tooth and four-tooth and alternate four-tooth standards for age estimation of 1200 Indian children aged 5-15 years old. DESIGN: The study was designed as a retrospective cross-sectional study. RESULTS: Demirjian's original seven-tooth method overestimated age by 0.64 ± 1.44, 0.75 ± 1.50 and 0.69 ± 1.46 years in boys, girls and the total sample, respectively. Demirjian's revised seven-tooth method overestimated age by + 0.24 ± 0.80, + 0.11 ± 0.81 and + 0.19 ± 0.80 years in boys, girls and the total sample, respectively. Demirjian's original four-tooth method overestimated age by 0.79 ± 1.59, 0.59 ± 2.77 and 0.72 ± 2.30 years in boys, girls and the total sample, respectively. Demirjian's alternate four-tooth method overestimated age by 1.31 ± 1.07, 1.20 ± 1.10 and 1.26 ± 1.08 years in boys, girls and the total sample, respectively. Statistically significant differences were observed between dental and chronological ages with all methods (p <0.001).  Significant gender-based differences were observed only with Demirjian's revised seven-tooth and original four-tooth methods (p <0.05).  Conclusion: The revised seven-tooth standards most accurately predicted the age of the study sample (mean prediction error = 2.28 months), followed by the original seven-tooth, four-tooth and alternate four-tooth standards. The Demirjian original seven-tooth method was significantly more accurate in boys compared to girls, while the reverse was true for the Demirjian revised seven-tooth and original four-tooth methods.

Erb's palsy complicating a case of annular pancreas.

We report an unusual presentation of annular pancreas with pneumoperitoneum in a newborn with an associated left sided Erb's palsy. The neurological deficit caused considerable confusion in the diagnosis and unexpected complications after surgery. We highlight the importance of clinical examination and the complications that an Erb's palsy can cause. This unusual triad of Erb's palsy, eventration of diaphragm and annular pancreas has hitherto not been described in literature.

A novel point mutation (L70P) inactivates poliovirus 3C protease.

Poliovirus (PV) contains a single-stranded positive-sense RNA genome, which is translated into a single polyprotein. Viral proteases process this polyprotein to produce several individual as well as fused proteins. The major viral protease 3C cleaves at nine of the eleven cleavage sites. During the process of expressing PV 3ABC protein in Escherichia coli, we identified a 3C mutant (L70P), which lost its protease activity. This loss of function was confirmed by generating recombinant adenoviruses expressing mutant and wild-type 3C. Further, infectious PV could not be recovered from PV full-length cDNA containing the L70P mutation. However, 3C L70P mutant cDNA could complement a PV cDNA containing a 1AB deletion, producing a viable virus population containing defective complementing genomes. Structural analysis of the mutant protein indicated that the L70P mutation resulted in the loss of a hydrogen bond between two residues located within a loop between two ß-sheets, potentially leading to strain on the catalytic site. We conclude that L70P inactivates 3C protease because of its close proximity to the 3C catalytic site.

Intracranial Perishunt Catheter Fluid Collections with Edema, a Sign of Shunt Malfunction: Correlation of CT/MRI and Nuclear Medicine Findings.

Fluid collections with edema along the intracranial tract of ventriculoperitoneal shunt catheters in adults are rare and are more frequently seen in children. The imaging appearance of these fluid collections is frequently confusing and presents a diagnostic dilemma. We present 6 cases of adult patients noted to have collections with edema along the tract of ventriculoperitoneal shunt catheters. To our knowledge, there are no previous studies correlating the CT/MR imaging findings with nuclear medicine scans in this entity. We hypothesized that when seen in adults, the imaging findings of a CSF-like fluid collection around the intracranial ventriculoperitoneal shunt catheter on CT/MR imaging may suggest areas of CSF accumulation with interstitial edema. It is important to recognize this rare ventriculoperitoneal shunt complication in adults to prevent misdiagnosis of an abscess or cystic tumor.

Bat Hunting and Bat-Human Interactions in Bangladeshi Villages: Implications for Zoonotic Disease Transmission and Bat Conservation.

Bats are an important reservoir for emerging zoonotic pathogens. Close human-bat interactions, including the sharing of living spaces and hunting and butchering of bats for food and medicines, may lead to spillover of zoonotic disease into human populations. We used bat exposure and environmental data gathered from 207 Bangladeshi villages to characterize bat exposures and hunting in Bangladesh. Eleven percent of households reported having a bat roost near their homes, 65% reported seeing bats flying over their households at dusk, and 31% reported seeing bats inside their compounds or courtyard areas. Twenty percent of households reported that members had at least daily exposure to bats. Bat hunting occurred in 49% of the villages surveyed and was more likely to occur in households that reported nearby bat roosts (adjusted prevalence ratio [aPR] 2.3, 95% CI 1.1-4.9) and villages located in north-west (aPR 7.5, 95% CI 2.5-23.0) and south-west (aPR 6.8, 95% CI 2.1-21.6) regions. Our results suggest high exposure to bats and widespread hunting throughout Bangladesh. This has implications for both zoonotic disease spillover and bat conservation.

Th17 Responses to Collagen Type V, kα1-Tubulin, and Vimentin Are Present Early in Human Development and Persist Throughout Life.

T helper 17 (Th17)-dependent autoimmune responses can develop after heart or lung transplantation and are associated with fibro-obliterative forms of chronic rejection; however, the specific self-antigens involved are typically different from those associated with autoimmune disease. To investigate the basis of these responses, we investigated whether removal of regulatory T cells or blockade of function reveals a similar autoantigen bias. We found that Th17 cells specific for collagen type V (Col V), kα1-tubulin, and vimentin were present in healthy adult peripheral blood mononuclear cells, cord blood, and fetal thymus. Using synthetic peptides and recombinant fragments of the Col V triple helical region (α1[V]), we compared Th17 cells from healthy donors with Th17 cells from Col V-reactive heart and lung patients. Although the latter responded well to α1(V) fragments and peptides in an HLA-DR-restricted fashion, Th17 cells from healthy persons responded in an HLA-DR-restricted fashion to fragments but not to peptides. Col V, kα1-tubulin, and vimentin are preferred targets of a highly conserved, hitherto unknown, preexisting Th17 response that is MHC class II restricted. These data suggest that autoimmunity after heart and lung transplantation may result from dysregulation of an intrinsic mechanism controlling airway and vascular homeostasis.

Hypertension is an important risk determinant for chronic kidney disease: results from a cross-sectional, observational study from a rural population in South India.

Hypertension (HTN) is an important cause for chronic kidney disease (CKD). There is a paucity of studies exploring the association of HTN with CKD from rural India. The objective of the study was to determine the prevalence and the determinants for HTN in a rural population in South India, focusing on the association with CKD. In a cross-sectional study on 2984 adults in three villages, demographic and anthropometric data were collected. Blood pressure was recorded on all individuals and prevalence was estimated. Proteinuria, microalbuminuria (MA) and estimated glomerular filtration rate (eGFR) were determined in a subgroup of 1331 subjects and the relation with HTN was analysed. Prevalence of HTN was 30.4% (95% CI 28.75-32.05%). In all, 78.08% had stage 1 HTN while 22% had stage 2 HTN. In total, 27.4% (249/908) subjects were aware of their hypertensive status. Of them, only 14.4% had adequate control of blood pressure. Age>50 years, diabetes, body mass index >25 and eGFR<60 ml min-1 were independent significant predictors of HTN (P<0.05). HTN was found to be an important risk factor for CKD even after adjusting for age, diabetes and cardiovascular risk factors (OR 2.22, 95% CI 1.46-3.36, P<0.001). The high prevalence of HTN, very low level of awareness and control, and the significant association with kidney disease indicate a need for stronger public health initiative with better penetration for awareness and screening for HTN and CKD in India's villages.

PDE11A negatively regulates lithium responsivity.

Lithium responsivity in patients with bipolar disorder has been genetically associated with Phosphodiesterase 11A (PDE11A), and lithium decreases PDE11A mRNA in induced pluripotent stem cell-derived hippocampal neurons originating from lithium-responsive patients. PDE11 is an enzyme uniquely enriched in the hippocampus that breaks down cyclic AMP and cyclic GMP. Here we determined whether decreasing PDE11A expression is sufficient to increase lithium responsivity in mice. In dorsal hippocampus and ventral hippocampus (VHIPP), lithium-responsive C57BL/6J and 129S6/SvEvTac mice show decreased PDE11A4 protein expression relative to lithium-unresponsive BALB/cJ mice. In VHIPP, C57BL/6J mice also show differences in PDE11A4 compartmentalization relative to BALB/cJ mice. In contrast, neither PDE2A nor PDE10A expression differ among the strains. The compartment-specific differences in PDE11A4 protein expression are explained by a coding single-nucleotide polymorphism (SNP) at amino acid 499, which falls within the GAF-B homodimerization domain. Relative to the BALB/cJ 499T, the C57BL/6J 499A decreases PDE11A4 homodimerization, which removes PDE11A4 from the membrane. Consistent with the observation that lower PDE11A4 expression correlates with better lithium responsiveness, we found that Pde11a knockout mice (KO) given 0.4% lithium chow for 3+ weeks exhibit greater lithium responsivity relative to wild-type (WT) littermates in tail suspension, an antidepressant-predictive assay, and amphetamine hyperlocomotion, an anti-manic predictive assay. Reduced PDE11A4 expression may represent a lithium-sensitive pathophysiology, because both C57BL/6J and Pde11a KO mice show increased expression of the pro-inflammatory cytokine interleukin-6 (IL-6) relative to BALB/cJ and PDE11A WT mice, respectively. Our finding that PDE11A4 negatively regulates lithium responsivity in mice suggests that the PDE11A SNPs identified in patients may be functionally relevant.

Salivary glucose as a diagnostic tool in Type II diabetes mellitus: A case-control study.

BACKGROUND AND OBJECTIVES: The prevalence of diabetes mellitus is increasing steadily in India. Understanding blood glucose level is the key to both diagnosis and management of diabetes mellitus. However, there is an on-going need for improvements in noninvasive, point-of-care tools for the diagnosis and prognosis of diabetes. Assessing a relationship between the blood glucose level and its concentration in other body fluids such as the saliva can help in developing a conservative method for blood sugar assessment replacing venous blood sampling. Diabetes mellitus is known to cause changes in salivary composition. Hence, this study was undertaken to evaluate the relationship of blood glucose level with salivary glucose in diabetic and nondiabetic patients. MATERIALS AND METHODS: The study sample included 100 diabetic patients and 100 nondiabetic patients aged above 35 years of age. Fasting blood and salivary glucose levels were measured in the two groups. Pearson's correlation coefficient was used to assess the correlation of blood glucose with salivary glucose in the two groups. RESULTS: The results of the study revealed an increase in the level of fasting salivary glucose in diabetics compared to that of nondiabetic patients. It also showed a highly significant positive correlation between fasting salivary glucose and serum glucose in both diabetic patients and in controls. CONCLUSION: From this study, it can be concluded that fasting salivary glucose level can be used as a noninvasive diagnostic, as well as a monitoring tool to assess the glycemic status of Type II diabetes mellitus patients.

Alkaptonuric arthropathy with periprosthetic fracture:a case report.

Alkaptonuria or Ochronosis is an autosomal recessive congenital metabolic error due to absence of enzyme homogentisic acid oxidase. Recently, we had an opportunity to diagnose this condition in one of our patients. An old aged man developed a pathological fracture of femur after a trivial fall at home. He was diagnosed to have intracapsular fracture neck of femur. Intraoperatively we noticed blackish discoloration of femoral head which lead us to investigate the case for alkaptonuria. Hemiarthroplasty was done with Austin moore prosthesis. Patient had cutaneous pigmentation and gross arthritic changes at multiple joints including spine. After one and half year patient developed a periprosthetic fracture of the femur for which open reduction and internal fixation was done. A periprosthetic fracture in ochronotic patient has never been reported till date which makes it a rarer entity. This article presents the clinico-radiologic manifestations of the case with surgical management and a follow up of two years.

Differential requirement for P2X7R function in IL-17 dependent vs. IL-17 independent cellular immune responses.

IL17-dependent autoimmunity to collagen type V (Col V) has been associated with lung transplant obliterative bronchiolitis. Unlike the T helper 1 (Th1)-dependent immune responses to Tetanus Toxoid (TT), the Th17 response to Col V in lung transplant patients and its Th1/17 variant observed in coronary artery disease patients requires IL-1ß, tumor necrosis factor α and CD14(+) cells. Given the involvement of the P2X7 receptor (P2X7R) in monocyte IL-1ß responses, we investigated its role in Th17-, Th1/17- and Th1-mediated proinflammatory responses. Transfer of antigen-pulsed peripheral blood mononucleated cells (PBMCs) from Col V-reactive patients into SCID mouse footpads along with P2X7R antagonists revealed a selective inhibition of Col V-, but not TT-specific swelling responses. P2X7R inhibitors blocked IL-1ß induction from monocytes, including both Col V-α1 peptide-induced (T-dependent), as well as native Col V-induced (T-independent) responses. Significantly higher P2X7R expression was found on CXCR3(neg) CCR4(+)/6(+) CD4(+) [Th17] versus CXCR3(+)CCR4/6(neg) CD4(+) [Th1] subsets in PBMCs, suggesting that the paradigm of selective dependence on P2X7R might extend beyond Col V autoimmunity. Indeed, P2X7R inhibitors suppressed not only anti-Col V, but also Th1/17-mediated alloimmunity, in a heart transplant patient without affecting anti-viral Epstein-Barr virus responses. These results suggest that agents targeting the P2X7R might effectively treat Th17-related transplant pathologies, while maintaining Th1-immunity to infection.