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Abstract Background A novel gradient, high-sensitive and specific stability-indicating reverse-phase HPLC method was developed and validated for quantitative purpose of known, unknown and degradant impurities profiling for diltiazem hydrochloride tablets. The impurities were separated on the Zorbax RX C8 column (150 mm × 4.6 mm, 5 μm) with mobile phase-A consisting of a mixture of 0.05 M sodium dihydrogen phosphate monohydrate buffer pH 3.0 and methanol in the ratio 800:200v/v and mobile phase-B consisting of acetonitrile with a flow rate of 1.0 mL min−1. The column compartment was maintained at 35 °C, and the detection wavelength was 240 nm. Diltiazem hydrochloride, its known impurities and unknown impurities have been well resolved from each other. Results The linearity of the method has been demonstrated across the concentration range of 0.18 to 5.65 µg mL−1 for EP impurity-F with correlation coefficient R 2 greater than 0.99. Recovery of method was proved from LOQ to 150% for known and unknown impurities with respect to test concentration and found in between 80 and 120%. Forced degradation study and specificity experiment results with mass balance proved the stability-indicating nature of the method and separated all known, unknown impurities and degradants from each other as well as from main drug component (diltiazem hydrochloride). The mass balance for stress study was found in between 95 and 105%. Conclusion Newly developed analytical method was validated as per ICH Q2 (R1) guidelines “Validation of analytical procedure” and found linear, accurate, specific, robust and precise in the established working range.

Abstract Background Pediatric leukemias have a diverse genomic landscape associated with complex structural variants, including gene fusions, insertions and deletions, and single nucleotide variants. Routine karyotype and fluorescence in situ hybridization (FISH) techniques lack sensitivity for smaller genomic alternations. Next-generation sequencing (NGS) assays are being increasingly utilized for assessment of these various lesions. However, standard NGS lacks quantitative sensitivity for minimal residual disease (MRD) surveillance due to an inherently high error rate. Methods Primary bone marrow samples from pediatric leukemia (n = 32) and adult leukemia subjects (n = 5), cell line MV4–11, and an umbilical cord sample were utilized for this study. Samples were sequenced using molecular barcoding with targeted DNA and RNA library enrichment techniques based on anchored multiplexed PCR (AMP®) technology, amplicon based error-corrected sequencing (ECS) or a human cancer transcriptome assay. Computational analyses were performed to quantitatively assess limit of detection (LOD) for various DNA and RNA lesions, which could be systematically used for MRD assays. Results Matched leukemia patient samples were analyzed at three time points; diagnosis, end of induction (EOI), and relapse. Similar to flow cytometry for ALL MRD, the LOD for point mutations by these sequencing strategies was ≥0.001. For DNA structural variants, FLT3 internal tandem duplication (ITD) positive cell line and patient samples showed a LOD of ≥0.001 in addition to previously unknown copy number losses in leukemia genes. ECS in RNA identified multiple novel gene fusions, including a SPANT-ABL gene fusion in an ALL patient, which could have been used to alter therapy. Collectively, ECS for RNA demonstrated a quantitative and complex landscape of RNA molecules with 12% of the molecules representing gene fusions, 12% exon duplications, 8% exon deletions, and 68% with retained introns. Droplet digital PCR validation of ECS-RNA confirmed results to single mRNA molecule quantities. Conclusions Collectively, these assays enable a highly sensitive, comprehensive, and simultaneous analysis of various clonal leukemic mutations, which can be tracked across disease states (diagnosis, EOI, and relapse) with a high degree of sensitivity. The approaches and results presented here highlight the ability to use NGS for MRD tracking.

Abstract The orphan small nucleolar RNA (snoRNA) ACA11 is overexpressed as a result of the t(4;14) chromosomal translocation in multiple myeloma (MM), increases reactive oxygen species, and drives cell proliferation. Like other snoRNAs, ACA11 is predominantly localized to a sub‐nuclear organelle, the nucleolus. We hypothesized that increased ACA11 expression would increase ribosome biogenesis and protein synthesis. We found that ACA11 overexpression in MM cells increased nucleolar area and number as well as argyrophilic nuclear organizing regions (AgNORs). Supporting these data, samples from t(4;14)‐positive patients had higher AgNORs scores than t(4;14)‐negative samples. ACA11 also upregulated ribosome production, pre‐47S rRNA synthesis, and protein synthesis in a ROS‐dependent manner. Lastly, ACA11 overexpression enhanced the response to proteasome inhibitor in MM cells, while no effect was found in response to high doses of melphalan. Together, these data demonstrate that ACA11 stimulates ribosome biogenesis and influences responses to chemotherapy. ACA11 may be a useful target to individualize the treatment for t(4;14)‐positive myeloma patients.

Acute leukemia represents the most common pediatric malignancy comprising diverse subtypes with varying prognosis and treatment outcomes. New and targeted treatment options are warranted for this disease. Patient-derived xenograft (PDX) models are increasingly being used for preclinical testing of novel treatment modalities. A novel approach involving targeted error-corrected RNA sequencing using ArcherDX HemeV2 kit was employed to compare 25 primary pediatric acute leukemia samples and their corresponding PDX samples. A comparison of the primary samples and PDX samples revealed a high concordance between single nucleotide variants and gene fusions whereas other complex structural variants were not as consistent. The presence of gene fusions representing the major driver mutations at similar allelic frequencies in PDX samples compared to primary samples and over multiple passages confirms the utility of PDX models for preclinical drug testing. Characterization and tracking of these novel cryptic fusions and exonal variants in PDX models is critical in assessing response to potential new therapies.

Aim: The aim is to review the English literature for post-traumatic superolateral dislocation of mandibular condyle (SDMC),discuss their dynamics and clinical management and to propose to modify the existing classification of SDMC. Patients and Methods: A literature search was carried at Pubmed, Sciencedirect, Google and references from reported articles were crosschecked to look for the cases of SDMC from 1969 to 2015 in English language. Also, we have reviewed 11 of our patients with total of 18 superolateral dislocated intact or sagittal split condyles ,who visited our unit in the previous two years. Results: In our retrospective analysis 58 cases of SDMC were found in the literature, of which 38 had intact mandibular condyles and 20 had sagittal split. Early and intact SDMC were successfully managed conservatively with closed reduction, whereas old cases and largely fractured condyles necessitated open reduction. Additionally, we observed an unusual dislocation associated with fracture of contralateral posterior mandible(angle) in our series which did not gratify the existing classification. Conclusion: Alteration of the existing classification was required to accommodate the unusual type of dislocation.

Background: Acute coronary syndrome (ACS) resulting from culprit lesion in left main coronary artery (LMCA) can cause rapid hemodynamic deterioration. It is important to identify these patients early to facilitate timely revascularization. ST segment elevation in aVR greater than or equal to V1 (aVR‐V1≥ 0) has been suggested as a sensitive predictor of LMCA disease. As a result of balanced forces, we hypothesized that ST deviation in V6 greater than or equal to ST deviation in V1 (V6‐V1≥ 0) might be a good determinant of LMCA disease. Methods: We compared admission 12‐lead ECGs of ACS resulting from culprit LMCA lesion (n = 75, group I) with ACS resulting from culprit left anterior descending lesion (n = 81, group II). Group I was selected over a period of 10 years. We compared V6‐V1≥ 0 to aVR‐V1≥ 0 in both groups. We also looked at ratios of ST deviations in V6,V1 (V6/V1≥ 1) and aVR,V1 (aVR/V1≥ 1) in patients where ST segment in V1 was not isoelectric (group I = 54 and group II = 55). Results: ST deviation in V6 was significantly greater in group I as compared to group II (P < 0.001). The reliabilities of V6‐V1≥ 0, V6/V1≥ 1, aVR‐V1≥ 0, and aVR/V1≥ 1 in predicting LMCA disease were determined. Conclusion: This is the largest series of ECG analysis on ACS resulting from culprit LMCA lesion. V6‐V1≥ 0 and V6/V1≥ 1 were more sensitive in predicting LMCA as culprit vessel in comparison to previously reported greater ST segment elevation in aVR than V1.

Dear Editor, Pancreatic glandular necrosis is the most common contributor to severe acute pancreatitis. When necrosis is present, the mortality and morbidity that come with acute pancreatitis are significantly higher, with associated infection in the necrotic area. Pancreatic fistulas generated by a ruptured pancreatic duct can develop following the surgery. In 15-23% of instances, pancreatic fistulas aggravate necrotising pancreatitis [1]. Treatment can range from early surgical debridement (“necrosectomy”) to aggressive intensive medical care [2]. This letter is about a 36-year-old male, who came to the casualty with complains of pain in abdomen radiating to his back and associated with distension of abdomen, nausea and 8-10 episodes of vomiting since five days. The patient was a chronic alcoholic. There were no other co-morbidities. He was admitted to the Intensive Medical Care Unit. On physical examination, the patient was afebrile, tachycardia was present, and abdomen was tender. Investigations revealed serum amylase to be 800 U/L, serum lipase 1400 U/L, total leucocyte count 12.2×109 /L, platelets 1.62 lac, and serum calcium 6mg/dl. Ultrasonography (abdomen and pelvis) was suggestive of acute pancreatitis, hepatomegaly with grade II fatty liver, and mild splenomegaly. The patient was kept nil by mouth and was started on fluid resuscitation, injection Meropenum 1 gm i/v thrice a day, injectable metronidazole 100 ml thrice a day, injection tramadol 100 ml i.v. thrice a day, pancreatin minimicrospheres capsules 25000-10000-10000 IU thrice a day, multivitamin with L-Methionine, and selenium yeast tablets twice a day. Contrast-enhanced CT abdomen revealed bulky pancreas throughout its course and peri-pancreatic fat stranding and fluid collection abutting lesser curvature of stomach suggestive of acute pancreatitis with necrotic collection, modified CT severity index score was 10. Interventional Radiologist’s opinion was taken in view of necrotic collection. An USG-guided catheter was inserted, and 300 ml output was obtained in five days. The drain was removed when there was less than 20 mL output per day. The patient developed anterior abdominal wall swelling within three days after the drain was removed [Table/Fig-1]. Local examination revealed pus draining through the catheter insertion site along with anterior abdominal wall swelling [Table/Fig-2]. As the abdominal distension increased CECT abdomen was repeated. It revealed acute pancreatitis with necrotic collection in subcutaneous and intramuscular plane of anterior abdominal wall, in the left hypochondriac and supra umbilical region communicating through a tract [Table/Fig-3]. The patient was planned for incision and drainage and 50 cc of necrotic material was drained [Table/Fig-4]. Describe patients condition before discharge and is awaiting followup after one month. This case reports an uncommon complication that happened during the treatment of a reasonably frequent acute surgical illness. Enteroatmospheric fistulas are known to have a notably higher fatality rate than their analogue, enterocutaneous fistulas [3-5]. Pancreaticoatmospheric fistulas may present in similar pattern and is associated with high mortality (28.6%) [3]. When compared to primary open necrosectomy, the Patients with Acute Necrotising Pancreatitis (PANTER) study found that the minimally invasive stepup method lowered the rate of complications and mortality. In this way, more than one-third of patients were effectively treated with percutaneous drainage only [6]. Management of severe acute necrotising pancreatitis necessitate multidisciplinary approaches, including resuscitative and supportive measures, as well as prompt management of fistula drainage for optimal wound management with eventual wound coverage and fistula closure.

Polyglobulia is secondary polycythaemia commonly due to underlying non haematological diseases like Chronic Obstructive Pulmonary Disease (COPD), obstructive sleep apnoea and also sometimes in people living in hilly or forest areas. It can occur in any venous or atrial thrombosis of the vessels, but cardiac, cerebral, and mesenteric vessels are usually involved. One of the rare causes of abrupt severe abdominal discomfort is portal vein thrombosis, usually associated with liver cirrhosis and thrombophilia. In this case report, the authors have highlighted a case of a 36-year-old male residing in the hilly area of Maharashtra, India, who reported to hospital with severe abdominal pain due to superior mesenteric vein thrombosis. On investigation, he had increased haemoglobin with raised haematocrit diagnosed as polyglobulia and became part of polycythaemia with positive JAK2 V617F mutation.

Tuberculosis (TB) is a multiorgan disease that can affect any part of the body. Though it is thought to be affecting mainly the pulmonary system, genitourinary TB is a rare but important manifestation of TB. It has been reported in