Atypical cellular responses mediated by intracellular constitutive active TrkB (NTRK2) kinase domains and a solely intracellular NTRK2-fusion oncogene.

Trk (NTRK) receptor and NTRK gene fusions are oncogenic drivers of a wide variety of tumors. Although Trk receptors are typically activated at the cell surface, signaling of constitutive active Trk and diverse intracellular NTRK fusion oncogenes is barely investigated. Here, we show that a high intracellular abundance is sufficient for neurotrophin-independent, constitutive activation of TrkB kinase domains. In HEK293 cells, constitutive active TrkB kinase and an intracellular NTRK2-fusion oncogene (SQSTM1-NTRK2) reduced actin filopodia dynamics, phosphorylated FAK, and altered the cell morphology. Atypical cellular responses could be mimicked with the intracellular kinase domain, which did not activate the Trk-associated MAPK/ERK pathway. In glioblastoma-like U87MG cells, expression of TrkB or SQSTM1-NTRK2 reduced cell motility and caused drastic changes in the transcriptome. Clinically approved Trk inhibitors or mutating Y705 in the kinase domain, blocked the cellular effects and transcriptome changes. Atypical signaling was also seen for TrkA and TrkC. Moreover, hallmarks of atypical pTrk kinase were found in biopsies of Nestin-positive glioblastoma. Therefore, we suggest Western blot-like immunoassay screening of NTRK-related (brain) tumor biopsies to identify patients with atypical panTrk or phosphoTrk signals. Such patients could be candidates for treatment with NTRK inhibitors such as Larotrectinhib or Entrectinhib.

SLAM (CD150) receptor homologous peptides block the peste des petits ruminants virus entry into B95a cells.

The fusion of haemagglutinin-neuraminidase (HN) protein of peste des petits ruminant (PPR) virus with signaling lymphocyte activation molecules (SLAM) host cell receptor consequences the virus entry and multiplication inside the host cell. The use of synthetic SLAM homologous peptides (i.e., molecular decoy for HN protein of PPR virus) may check PPR infection at the preliminary stage. Hence, the predicted SLAM homologous peptides using bioinformatics tools were synthesized by solid phase chemistry with standard Merrifield's 9-fluorenylmethoxycarbonyl (Fmoc) chemistry and were purified by reverse phase high performance liquid chromatography. The secondary structures of synthesized peptides were elucidated by circular dichroism spectroscopy. The in vitro interactions of these peptides were studied through indirect Enzyme Linked Immuno Sorbent Assay (ELISA) and visual surface plasmon UV-visible spectroscopy. The SLAM homologous peptides were able to interact with the peste des petits ruminant virus (PPRV) with varying binding efficiency. The interaction of SLAM homologous peptide with the PPR virus was ascertained by the change in the plasmon color from red wine to purple during visual detection and also by bathochromic shift in absorbance spectra under UV-visible spectrophotometry. The cytotoxic and anti-PPRV effect of these peptides were also evaluated in B95a cell line using PPR virus (Sungri/96). The cytotoxic concentration 50 (CC50 ) value of each peptide was greater than 1000 µg mL-1 . The anti-PPRV efficiency of SLAM-22 was relatively high among SLAM homologous peptides, SLAM-22 at 25 µg mL-1 concentration showed a reduction of more than log10 3 virus titer by priming of B95a cell line while the use of SLAM-15 and Muco-17 at the same concentration dropped virus titer from log10 4.8 to log10 2.5 and log10 3.1 respectively. The concentration of SLAM homologous peptide (25 µg mL-1 ) to exert its anti-PPRV effect was much less than its CC50 level (>1000 µg mL-1 ). Therefore, the synthetic SLAM homologous peptides may prove to be better agents to target PPRV.

Correlation Between Carotid Intima Media Thickness and Non-Alcoholic Fatty Liver Disease.

CONTEXT: Non-alcoholic fatty liver disease (NAFLD) is considered a potential independent risk factor for cardiovascular disease. Increased carotid intima-media thickness (CIMT) is a sign of early atherosclerosis and is linked to an increased risk of myocardial infarction, stroke, and peripheral vascular disease. AIM: To study correlation between CIMT and NAFLD and its association with increase in grades of NAFLD. Material and Study Design: An observational case control study of 40 cases and 40 controls (age and sex matched) was done. The difference of CIMT between the two groups was analysed. CIMT was also measured among the various grades of NAFLD cases. MATERIALS AND METHODS: 40 cases with NAFLD and 40 controls falling within the age group of 18-45 years were taken in the study. Patients with history of significant alcohol consumption, acute or chronic liver disease, diabetes mellitus, hypertension, malignancy, hypothyroidism and having dyslipidaemia, CAD and stroke were excluded from the study. All the subjects underwent abdominal and carotid ultrasound in order to assess NAFLD and CIMT measurement. The left and right common carotid artery was examined using PHILIPS HD11XE high-definition ultrasound system equipped with a 3-12 MHz linear array transducer in B mode. OBSERVATION AND RESULTS: There was a statistically significant difference between the 2 groups in terms of CIMT with a p value of <0.001. The mean CIMT in the Case group was 0.86 mm while in Control group was 0.52 mm. There was a significant difference between the 3 Grades of NAFLD in terms of CIMT with a p value of <0.001 with maximum CIMT being in Grade 3 of NAFLD. Body Mass Index, Alanine Transaminase, Aspartate Transaminase, Alkaline Phosphatase, Total Cholesterol, Triglycerides, Low Density Lipoprotein were also found to have statistical significant difference between cases and controls. Age, gender, Blood pressure, Fasting Blood Sugar, HbA1c, Hemoglobin, Total leucocyte Count, Platelet count, Serum Bilirubin, Total protein and Albumin were found to be statistically insignificantly different between cases and controls. CONCLUSION: CIMT is increased in NAFLD patients. Increase in CIMT is significantly correlated with increasing grades of NAFLD. Hence CIMT can be used as screening tests in NALFD patients to assess cardiovascular risks.

Homeostatic calcium fluxes, ER calcium release, SOCE, and calcium oscillations in cultured astrocytes are interlinked by a small calcium toolkit.

How homeostatic ER calcium fluxes shape cellular calcium signals is still poorly understood. Here we used dual-color calcium imaging (ER-cytosol) and transcriptome analysis to link candidates of the calcium toolkit of astrocytes with homeostatic calcium signals. We found molecular and pharmacological evidence that P/Q-type channel Cacna1a contributes to depolarization-dependent calcium entry in astrocytes. For stimulated ER calcium release, the cells express the phospholipase Cb3, IP3 receptors Itpr1 and Itpr2, but no ryanodine receptors (Ryr1-3). After IP3-induced calcium release, Stim1/2 - Orai1/2/3 most likely mediate SOCE. The Serca2 (Atp2a2) is the candidate for refilling of the ER calcium store. The cells highly express adenosine receptor Adora1a for IP3-induced calcium release. Accordingly, adenosine induces fast ER calcium release and subsequent ER calcium oscillations. After stimulation, calcium refilling of the ER depends on extracellular calcium. In response to SOCE, astrocytes show calcium-induced calcium release, notably even after ER calcium was depleted by extracellular calcium removal in unstimulated cells. In contrast, spontaneous ER-cytosol calcium oscillations were not fully dependent on extracellular calcium, as ER calcium oscillations could persist over minutes in calcium-free solution. Additionally, cell-autonomous calcium oscillations show a second-long spatial and temporal delay in the signal dynamics of ER and cytosolic calcium. Our data reveal a rather strong contribution of homeostatic calcium fluxes in shaping IP3-induced and calcium-induced calcium release as well as spatiotemporal components of intracellular calcium oscillations.

Insights of 8-hydroxyquinolines: A novel target in medicinal chemistry.

8-Hydroxyquinoline (8-HQ) is a significant heterocyclic scaffold in organic and analytical chemistry because of the properties of chromophore and is used to detect various metal ions and anions. But from the last 2 decades, this moiety has been drawn great attention of medicinal chemists due to its significant biological activities. Synthetic modification of 8-hydroxyquinoline is under exploration on large scale to develop more potent target-based broad spectrum drug molecules for the treatment of several life-threatening diseases such as anti-cancer, HIV, neurodegenerative disorders, etc. Metal chelation properties of 8-hydroxyquinoline and its derivatives also make these potent drug candidates for the treatment of various diseases. This review comprises 8-hydroxyquinoline derivatives reported in the literature in last five years (2016-2020) and we anticipate that it will assist medicinal chemists in the synthesis of novel and pharmacologically potent agents for various therapeutic targets, mainly anti-proliferative, anti-microbial, anti-fungal and anti-viral as well as for the treatment of neurodegenerative disorders.

Giant metastasizing malignant hidradenoma in a child.

An 8-year-old girl presented with a scalp swelling. The swelling was recurrent, reappearing everytime after local excision. She underwent surgery and the histopathologic diagnosis was malignant hidradenoma. This very rare and aggressive tumor is known to occur in elderly population and is histopathologically distinct from its commonly occuring benign counterpart. Malignant hidradenoma is resistant to chemotherapy and radiotherapy. We empahsize that being cognizant of the possibility of this rare tumor would assist in timely action in the form of wide resection, with possible reduction in morbidity and mortality.

Magnetic resonance imaging - A troubleshooter in obstetric emergencies: A pictorial review.

The application of magnetic resonance imaging (MRI) in pregnancy faced initial skepticism of physicians because of fetal safety concerns. The perceived fetal risk has been found to be unwarranted and of late, the modality has attained acceptability. Its role in diagnosing fetal anomalies is well recognized and following its safety certification in pregnancy, it is finding increasing utilization during pregnancy and puerperium. However, the use of MRI in maternal emergency obstetric conditions is relatively limited as it is still evolving. In early gestation, ectopic implantation is one of the major life-threatening conditions that are frequently encountered. Although ultrasound (USG) is the accepted mainstay modality, the diagnostic predicament persists in many cases. MRI has a role where USG is indeterminate, particularly in the extratubal ectopic pregnancy. Later in gestation, MRI can be a useful adjunct in placental disorders like previa, abruption, and adhesion. It is a good problem-solving tool in adnexal masses such as ovarian torsion and degenerated fibroid, which have a higher incidence during pregnancy. Catastrophic conditions like uterine rupture can also be preoperatively and timely diagnosed. MRI has a definite role to play in postpartum and post-abortion life-threatening conditions, e.g., retained products of conception, and gestational trophoblastic disease, especially when USG is inconclusive or inadequate.

Immune Response to Double Dose Hepatitis-B Vaccine using Four Dose Schedule in HIV Infected Children.

OBJECTIVE: To assess the immune response to hepatitis B vaccination in HIV infected children using 4 double dose schedule administered at 0-1-2-6 mo. METHODS: In a prospective observational study, 40 HIV infected children were vaccinated with hepatitis B virus (HBV) vaccine (20 mcg, 1 ml IM at anterolateral aspect of thigh of recombinant DNA vaccine) at 0-1-2-6 mo. Anti-HBsAb titre were assessed 4-8 wk after the last dose. Clinical severity was assessed according to WHO staging. Immune status of the patients was assessed using CD 4+ counts before the vaccination. RESULTS: Of the total 40 patients, 33 (28 boys, 5 girls) completed the study. Six patients were in pre-antiretroviral therapy (ART) care while 27 were receiving ART for a mean duration of 2.06 y. Ten patients belonged to WHO clinical stage 1, and 17, 2 and 4 patients to WHO clinical stage 2, 3 and 4 respectively. Median CD4+ cell count was 738/mm(3) and 28 patients had mild or no immunosuppression. Out of the total 33 patients who completed followup, only 2 patients (6 %) did not seroconvert (Anti HBsAb titre at end of study <10 IU/L), the rest (94 %) achieved different levels of Anti HBsAb titre at end of study. Twenty two (66 %) patients had anti HBsAb titre more than 1000 IU/L, 8 (24 %) had titre between 100 and 1000 IU/L and one (3 %) patient had level of 10-99.99 IU/L. CONCLUSIONS: In HIV-infected children who have no or mild immunosuppression, four dose, double dose schedule of HBV vaccine achieves very high seroconversion rates.

Fungal production of single cell oil using untreated copra cake and evaluation of its fuel properties for biodiesel.

This study evaluated the microbial conversion of coconut oil waste, a major agro-residue in tropical countries, into single cell oil (SCO) feedstock for biodiesel production. Copra cake was used as a low-cost renewable substrate without any prior chemical or enzymatic pretreatment for submerged growth of an oleaginous tropical mangrove fungus, Aspergillus terreus IBB M1. The SCO extracted from fermented biomass was converted into fatty acid methyl esters (FAMEs) by transesterification and evaluated on the basis of fatty acid profiles and key fuel properties for biodiesel. The fungus produced a biomass (8.2 g/l) yielding 257 mg/g copra cake SCO with ~98% FAMEs. The FAMEs were mainly composed of saturated methyl esters (61.2%) of medium-chain fatty acids (C12-C18) with methyl oleate (C18:1; 16.57%) and methyl linoleate (C18:2; 19.97%) making up the unsaturated content. A higher content of both saturated FAMEs and methyl oleate along with the absence of polyunsaturated FAMEs with ≥4 double bonds is expected to impart good fuel quality. This was evident from the predicted and experimentally determined key fuel properties of FAMEs (density, kinematic viscosity, iodine value, acid number, cetane number), which were in accordance with the international (ASTM D6751, EN 14214) and national (IS 15607) biodiesel standards, suggesting their suitability as a biodiesel fuel. The low cost, renewable nature, and easy availability of copra cake, its conversion into SCO without any thermochemical pretreatment, and pelleted fungal growth facilitating easier downstream processing by simple filtration make this process cost effective and environmentally favorable.

Metanephric stromal tumor: An unusual pediatric renal neoplasm.

A renal tumor in a 14-month- old child, who was initially diagnosed as mesoblastic nephroma, but on review post surgery was diagnosed as hyper-differentiated metanephric stromal tumor, with its excellent prognostic outcome. An attempt is made to document imaging features that may enable one to suspect this rare condition. The literature is reviewed with emphasis on its distinction from its look-alikes in the pediatric age group.

Small bowel tuberculosis by multidetector CT enteroclysis.

PURPOSE: To prospectively evaluate the role of multidetector computed tomography (CT) enteroclysis in evaluation and diagnostic characterization of suspected small bowel tuberculosis. MATERIALS AND METHODS: The study group included 21 human subjects suspected of having small bowel tuberculosis. A nasoenteric tube was positioned into the duodenojejunal junction under fluoroscopic guidance and water was infused by hand injection. After intravenous administration of 120 mL of iodinated contrast material, multidetector CT enteroclysis images were obtained with 40 × 0.625 mm collimation and findings were analyzed by three readers working in consensus. Findings were compared with the results of endoscopy, histopathological analysis, and clinical follow up. RESULTS: Multidetector CT enteroclysis was well tolerated in 20 patients; one of the patients complained of vomiting during intubation. Using multidetector CT enteroclysis diagnosis of abdominal tuberculosis was made in sixteen patients, out of which small bowel involvement was seen in 13 patients. Multidetector CT enteroclysis demonstrated Crohn's disease in one patients and normal in four patients. The sensitivity and specificity in diagnosing abdominal tuberculosis by CT enteroclysis came out to be 93.75 and 100 % respectively. CONCLUSIONS: Multidetector CT enteroclysis is well tolerable and accurate modality for evaluation and diagnostic characterization of small bowel tuberculosis.

OHVIRA: Uterus didelphys, blind hemivagina and ipsilateral renal agenesis: Advantage MRI.

We present here a case of an uncommon complex uterine anomaly - Obstructed HemiVagina with Ipsilateral Renal Agenesis (OHVIRA), also known as Herlyn-Werner-Wunderlich syndrome in a 14-year-old girl along with sonographic (trans-abdominal and trans labial), and MRI findings. The patient underwent surgery wherein imaging findings were confirmed. An MRI has proved to be of great help in correct diagnosis avoiding surgical interventions/ laparoscopy, which were needed in past to diagnose this rare anomaly. We also discuss the development of this anomaly with the help of a relatively new theory of uro-genital development by Acien and review the literature.

Occupational exposure to human immunodeficiency virus in health care providers: a retrospective analysis.

OBJECTIVES: To determine the population at risk, risk factors, and outcome of occupational exposure to blood and body fluids in health care providers. MATERIALS AND METHODS: Retrospective review of two and half year data of ongoing surveillance of occupational exposure to blood and body fluids in a tertiary care hospital. RESULTS: 103 Health Care Providers (HCP) reported an occupational exposure to blood and body fluids during the period under review. These comprised 72 (69.9%) doctors, 20 (19.4%) nursing personnel, and 11 (10.6%) cleaning staff. Of the doctors, 65% were interns. 53.4% HCP had work experience of less than one year. Circumstances of exposure included clinical procedures (48%), sweeping/handling used sharps (29%), recapping (16%), and surgery (6.9%). 74.3% of the exposures were due to non-compliance with universal precautions and were thus preventable. The device most frequently implicated in causing injury was hollow bore needle (n=85, 82.5%). Human Immunodeficiency Virus (HIV) status of the source was positive in 6.8% cases, negative in 53.4% cases, and unknown in remaining 39.8% cases. Postexposure prophylaxis (PEP) was indicated in 100 (97.08%) cases and was initiated within 2 h of exposure in 26.8% HCP. In 23.2% HCP, PEP initiation was delayed beyond 72 h of exposure due to late reporting. Thirteen HCP received expanded and the remaining received basic regime. Of the 82 HCP followed up, 15 completed the full course, while 55 stopped PEP after the first dose due to negative source status. Twelve HCP with exposure to blood of unknown HIV status discontinued PEP despite counseling. Complete follow-up for seroconversion was very poor among the HCP. HIV status at 6 month of exposure is not known for any HCP. CONCLUSIONS: Failure to follow universal precautions including improper disposal of waste was responsible for majority of occupational exposures. HCP need to be sensitized regarding hospital waste management, management of occupational exposure, need for PEP, and continued follow-up.

Bone marrow-derived mesenchymal stromal cells enhance chimeric vessel development driven by endothelial cell-coated microtissues.

Adding bone marrow-derived mesenchymal stromal cells (bmMSCs) to endothelialized collagen gel modules resulted in mature vessel formation, presumably caused in part by the observed display of pericyte-like behavior for the transplanted GFP(+) bmMSCs. A previous study determined that rat aortic endothelial cells (RAECs) delivered on the surface of small (∼0.8 mm long×0.5 mm diameter) collagen gel cylinders (microtissues, modular tissue engineering) formed vessels after transplantation into immunosuppressed Sprague-Dawley (SD) rats. Although the RAECs formed vessels in this allogeneic transplant model, there was a robust inflammatory response and the vessels that formed were leaky as shown by microcomputed tomography (microCT) perfusion studies. In vitro assays showed that SD rat bmMSCs embedded into the collagen gel modules increased the extent of EC proliferation and enhanced EC sprouting. In vivo, although vessel number was not affected, the new vessels formed by the bmMSCs and RAECs were more stable and leaked less in the microCT perfusion analysis than vessels formed by implanted RAECs alone. Addition of the bmMSCs also decreased the total number of CD68(+) macrophages that infiltrated the implant and changed the distribution of CD163(+) (M2) macrophages so that they were found within the newly developed vascularized tissue. Most interestingly, the bmMSCs became smooth muscle actin positive and migrated to surround the EC layer of the vessel, which is the location typical of pericytes. The combination of these two effects was presumed to be the cause of improved vascularity when bmMSCs were embedded in the EC-coated modules. Further exploration of these observations is warranted to exploit modular tissue engineering as a means of forming large vascularized functional tissues using microtissue components.

Case report: Subutaneous hemangiomatosis causing Kasabach-Merritt syndrome - MRI features.

Hemangiomatosis is an uncommon entity in which there is diffuse infiltration of soft tissue or bone by hemangioma. Kasabach-Merritt syndrome is an uncommon complication of large hemangiomas, in which there is thrombocytopenia and coagulopathy. Plain radiographs, in addition to showing a soft tissue mass, also show a variety of findings in the bones. MRI is the investigation of choice. A case of a 2-year-old child suffering from hemangiomatosis and a resultant Kasabach-Merritt syndrome is presented.

Unilateral renal lymphangiectasia: imaging appearance on sonography, CT and MRI.

Lymphangiomas are benign tumors of unknown etiology often seen in children, involving the neck (75%), and axillary (20%) region, but rarely the kidney [Zapzalka et al. (2002) Lymphangioma of the renal capsule. J Urol 168:220.]. We report a case of unilateral renal lymphangiectasia, where the diagnosis was made on imaging.