Design and synthesis of Meldrum's acid based 7-azaindole anchored 1,2,3-triazole hybrids as anticancer agents.

A series of Meldrum's acid, 7-azaindole and 1,2,3-triazole hybrids were synthesized and evaluated for their in vitro anticancer activity against five different cancer cell lines viz. MCF-7 (breast cancer), HeLa (cervical cancer), DU-145 (prostate cancer), HepG2 (liver cancer) and K562 (myelogenous leukemia cell). Among the series, compound 6b containing a 4-methyl substitution showed potent activity against HeLa cell line. Cell cycle analysis revealed that compound 6b induced cell cycle arrest at the G2/M phase and induced apoptosis. Apoptotic activity was further confirmed by Hoechst staining and Annexin V-FITC assay. Compound 6b has been found to exhibit higher activity in all four cell lines, with IC50 values of 6.67 ± 0.39 µM, 4.44 ± 0.32 µM, 12.38 ± 0.51 µM and 9.97 ± 0.25 µM against MCF-7, HeLa, DU-145 and HepG2 cell lines respectively. Compounds 6m (9.68 ± 0.10 µM) and 6n (9.52 ± 0.38 µM), which have dimethoxy and trimethoxy substitutions, respectively, have demonstrated significant anticancer activity against HeLa cells compared to the other cells. The molecular docking study of ligand 6b against the crystal structure of EGFR and Mcl-1 scored notable binding energy values and displayed important interactions like H-bond, π-cation and other hydrophobic interactions.

The core four - A panel of immunohistochemistry markers to diagnose and subtype testicular germ cell tumors.

CONTEXT: Immunohistochemistry (IHC) to differentiate germ cell tumors. AIMS: The aim of the study is to differentiate seminomatous and nonseminomatous germ cell tumors (GCTs) with morphological overlap using a minimal and affordable panel of IHC markers. SETTINGS AND DESIGN: This is a retrospective observational study. SUBJECTS AND METHODS: All testicular GCTs (TGCT) which were diagnosed on biopsies and/or resection specimens (prechemotherapy) between January 2014 and June 2019. The demographic, clinical, and imaging findings were noted from the medical records. Hematoxylin and eosin (H and E)-stained sections were reviewed for morphology. The IHC markers constituted Octamer-binding transcription factor (OCT) 3/4, glypican 3 (GPC3), CD117, CD30, placental-like alkaline phosphatase, Sal-like protein 4, and ß-human chorionic gonadotropin (HCG). IHC markers were performed in various combinations depending on the morphology, and a panel constituting OCT 3/4, CD117, GPC3, and CD30 was performed on cases with diagnostic dilemma and morphological overlaps. STATISTICAL ANALYSIS USED: Sensitivity, specificity, positive (PPV), and negative predictive value (NPV) were calculated for suggested panel of IHC OCT 3/4, CD117, GPC3, and CD30. RESULTS: The study included 36 patients with TGCT with a mean age of 27 (15-58) years. Nonseminomatous tumors were the most common (86%). The concise panel was performed in 20/36 (56%) tumors to resolve the diagnosis. The sensitivity, specificity, PPV, and NPV for OCT3/4 were 80%, 55%, 31%, and 92% in seminomas and 65%, 100%, 100%, and 46% in embryonal carcinomas (EC), for CD117 was 89%, 82%, 73%, and 93% in seminomas and 60%, 77%, 60%, and 77% in yolk sac tumors (YST), for GPC3 was 95%, 90%, 95%, and 90% in YST, CD30 96%, 100%, 100%, and 91% in ECs, respectively. CONCLUSIONS: Designing a novel concise and affordable IHC panel constituting OCT 3/4, CD117, GPC3, and CD30 has good sensitivity and specificity in differentiating seminomas, YST, and EC, respectively. Additional markers, namely ß-HCG, can be used in identifying the choriocarcinoma component.

Quantification of sweat urea in diabetes using electro-optical technique.

OBJECTIVE: To develop a device for quantifying sweats urea concentration. APPROACH: The proposed optical device uses light source, light sensor with time and intensity controlled operation. Sweat samples are collected from a group of volunteers belonging to control and diabetes. After sedimentation and suitable pre-processing, sweat samples are irradiated by primary colour light sources operated sequentially. Reflected light intensity is used to compute the sweat urea concentration. At first the test is performed using aqueous solution with known urea concentration. After ascertaining the correctness of the device, the same test procedure is repeated for sweat samples collected from 90 volunteers (30 per group) belonging to controls, type II diabetes less than 5 years and type II diabetes for more than 5 years. MAIN RESULTS: Computed urea concentration when compared with standard lab techniques like UV-visible absorption spectroscopy and colorimeter, a correlation of 98% with error less than 3% is achieved. The results also demonstrate an elevation in sweat urea level with years of diabetes, in spite of the serum urea level being within limits. We extended the study on a few kidney disease subjects and observed the influence of blood glucose on urea. SIGNIFICANCES: Diabetic kidney disease is one result of prolonged elevation in blood glucose levels. When insulin secretion reduces, the serum urea level increases and vice versa is also true. Hence, monitoring urea level in blood is important in diabetic subjects. Any change in serum urea will have an impact on sweat urea concentration. Therefore, the proposed device can be used to measure sweat urea periodically, so that any change can be observed at an early stage and diabetic nephropathy could be prevented at large.

GATA-3 Expression in all Grades and Different Variants of Primary and Metastatic Urothelial Carcinoma.

Urothelial carcinoma has a varied and wide histological spectrum posing a diagnostic challenge in H&E examination alone. Immunohistochemical markers like GATA-3 along with other appropriate panel of IHC can be used. However, the percentage positivity and its intensity may vary in different variants and grades of primary and metastatic urothelial carcinoma. To observe the GATA-3 expression patterns in all the grades and different variants of primary and metastatic urothelial carcinomas. It is a prospective and retrospective observational study. All the clinically suspected urothelial carcinoma (UC) during January 2016 to December 2017 were included in the study. Depending on the differential diagnosis considered, immunohistochemistry (IHC) markers including CK7, CK20, p63, AMACR, CDX2, and p16 were done to differentiate UC from other primary carcinomas. The tumors confirmed as UC were analyzed further for GATA-3 expression by Chi-square test. The number of UC in the present study was 126 including 122 (bladder in 107, ureter in 7, renal pelvis in 5, and urethra in 3) primary and 4 metastatic UC (3 in lung and 1 in liver). Age of the patients ranged from 29 to 80 (mean 61.28) years with male/female ratio 4:1. GATA-3 showed positivity in 97 (79.5%) primary UC. GATA-3 was positive in all normal urothelium and non-invasive UC (100%), while it was positive in 69/94 (73.4%) invasive UC including variants. GATA-3 was positive in 35/39 LP invasive (89.74%) and 34/55 (61.81%) MP invasive UC. GATA-3 was positive in 39/40 papillary cases (97.5%) and 45/59 (76.27%) cases of non-papillary UC. GATA-3 showed strong expression in all metastatic UC (100%). GATA-3 expression was seen in 101/126 (80.15%) of UC including primary and metastatic carcinomas and hence was a useful marker in diagnosing UC. The GATA-3 positivity decreased from normal urothelium to UC; low-grade UC to high-grade UC; non-invasive to invasive UC; lamina propria invasive to muscle invasive UC; papillary to non-papillary UC.

Comparison of Clinician Versus Parental Perspectives of Outcomes in Cochlear Implantees: A South Indian Experience.

Outcomes of cochlear implantation (CI) are generically assessed using standard validated measures like CAP, SIR, MAIS and MUSS scales. Although this reflects the improvement in auditory verbal skills among the implantees with habilitation over one year, the overall perception of their skill development may vary between the parents of these children and the clinicians who provide the habilitation. This study aimed to compare the CAP and SIR scores sequentially over habilitation and further analyzes the correlation between clinician assessment (with CAP/SIR scores) and parental perspective (with MAIS/MUSS scores), at the end of one year of habilitation. 388 children aged 1-6 years who underwent unilateral CI were included in the study. Their baseline CAP and SIR scores were recorded post implantation. All children received 1 year of intensive auditory verbal therapy and their 12 month CAP, SIR, MAIS and MUSS scores were then recorded. The baseline CAP/SIR scores were compared with 12 month CAP/SIR scores and then their 12 month CAP/SIR scores were correlated with 12 month MAIS/MUSS scores respectively. There was significant difference between baseline and the 12 month CAP/SIR scores (p < 0.001). There was strong positive correlation between CAP and SIR scores after 12 months of habilitation (r = 0.7), while there was moderate positive correlation between CAP and MAIS scores (r = 0.59) and between SIR and MUSS scores (r = 0.49) respectively. Though the parents note significant improvement in child's communication abilities, the parental perspective of final outcomes does not always match with the clinician's assessments at the end of habilitation, as highlighted by the moderate correlations. A more precise method of holistic assessment is lacking currently and stands warranted.

Diagnostic utility of immunohistochemical marker prostein for evaluation of primary and metastatic prostatic carcinomas.

CONTEXT: The diagnosis of prostatic adenocarcinoma on histopathology depends on architectural and cytomorphological features supported by immunohistochemistry (IHC). Though all the prostate markers show excellent specificity, the sensitivity and percentage positivity vary. AIMS: In this study, we aim to study the expression of prostein in normal, benign, and malignant (primary and metastatic) lesions with particular emphasis on its utility in the differential diagnosis of poorly differentiated and metastatic prostatic adenocarcinoma along with a standard panel of IHC markers. SETTINGS AND DESIGN: This was both a prospective and retrospective as well as descriptive and observational study. SUBJECTS AND METHODS: All samples from patients with clinically suspected carcinoma prostate from both primary and metastatic sites from June 2015 to May 2016 were included in the study. Samples with difficulty in diagnosis on hematoxylin and eosin staining were subjected to a panel of IHC markers along with prostein. STATISTICAL ANALYSIS USED: Receiver operating curve analysis and Chi-square test. RESULTS: Prostein showed a 100% sensitivity and specificity to identify normal prostatic epithelium, benign and premalignant lesions, and prostatic adenocarcinoma. Prostein showed a specificity of 100% in differentiating prostatic carcinoma from poorly differentiated urothelial carcinoma and in differentiating metastatic prostatic carcinoma from adenocarcinoma of nonprostatic origin. CONCLUSIONS: Prostein is a new and promising prostate-specific marker that showed slightly more sensitivity and specificity than prostate-specific antigen. Thus, adding prostein to the IHC panel will greatly improve the detection of poorly differentiated primary and metastatic lesions of the prostate.

Predicting the molecular subtype of breast cancer based on mammography and ultrasound findings.

AIM: To determine the correlation between mammography and ultrasound features of breast cancer with molecular subtypes and to calculate the predictive value of these features. MATERIALS AND METHOD: This is a prospective study of consecutive patients with breast cancer presenting between January 2016 and July 2017, who underwent mammography and/or ultrasound of breast and excision of the breast mass. Patients with contralateral breast mass, metastases, h/o prior cancer treatment, and other malignancies were excluded. On mammography, the presence or absence of microcalcification was noted. On ultrasound examination size, margins, microcalcification, posterior acoustic features, vascularity, and axillary nodes were assessed. Margins were categorized into circumscribed and non-circumscribed. Posterior acoustic features were classified into four categories: shadowing, enhancement, mixed, and no changes. Vascularity was assessed based on Adler's index into grades 0, 1, 2, and 3. Grades 0 and 1 were considered low and 2 and 3 were high. RESULTS: Tumors with non-circumscribed margins and posterior acoustic shadowing were likely to be luminal A or B subtype of breast cancer [odds ratio (OR) 5.78; 95% confidence interval (CI) 3.68-9.80; P < 0.0001]. Tumors with non-circumscribed margins, posterior acoustic shadowing, and high vascularity were more likely to be luminal B subtype (OR 2.88; 95% CI 2-4.14; P- <0.0001). Tumors with microcalcification and posterior mixed acoustic pattern were strongly associated to be HER2-positive (OR 5.48; 95% CI 3.06-9.80; P < 0.0001). Tumors with circumscribed margins and posterior acoustic enhancement were highly suggestive of triple-negative breast cancer (OR 7.06; 95% CI 4.64-10.73; P < 0.0001). CONCLUSION: Microcalcification detected on mammography and certain ultrasound features such as circumscribed or non-circumscribed margins, posterior acoustic features, and vascularity are strongly correlated in predicting the molecular subtypes of breast cancer, and thus may further expand the role of conventional breast imaging.

Convolutional Neural Network for Segmentation and Measurement of Intima Media Thickness.

The measurement of Carotid Intima Media Thickness (IMT) on Common Carotid Artery (CCA) is a principle marker of risk of cardiovascular disease. This paper presents a novel method of using deep Convolutional Neural Network (CNN) for identification and measurement of IMT on the far wall of the artery. The Region of Interest (ROI) is extracted using CNN architecture with 8 layers. 110 subjects are taken for the study. Each subject is recorded with one Right Common Carotid Artery (RCCA) and Left Common Carotid Artery (LCCA) frame resulting in 220 recordings. Patch based segmentation with 2640 patches are given to the training network for ROI localization. Intima Media Complex (IMC) is the area where IMT is measured. This region is extracted after defining the ROI. Keeping in mind the end objective of measurement of IMT values binary threshold with snake algorithm is applied to extract the lumen-intima and media-adventitia boundary. IMT values are measured for 20 cases and mean difference is found to be 0.08 mm.

A Case of AML-M2 with Sole Interstitial Deletion in 9q Without AML1-ETO/Inv 16 Rearrangement and FLT3/NPMI Mutations.

Conventional/molecular cytogenetics is important in identification of genomic abnormalities, for prognostication and in risk stratification of de novo patients with acute myeloid leukemias (AML). Here we present an AML M2 case showing the sole karyotypic abnormality, the rare interstitial deletion in the long arm of chromosome 9 with the loss of segment q12-q13.

Evaluation of antioxidant, antibacterial and cytotoxic effects of green synthesized silver nanoparticles by Piper longum fruit.

Silver nanoparticles synthesized through bio-green method has been reported to have biomedical applications to control pathogenic microbes as it is cost effective compared to commonly used physical and chemical methods. In present study, silver nanoparticles were synthesized using aqueous Piper longum fruit extract (PLFE) and confirmed by UV-visible spectroscopy. The nanoparticles were spherical in shape with an average particle size of 46nm as determined by scanning electronic microscopy (SEM) and dynamic light scattering (DLS) particle size analyzer respectively. FT-IR spectrum revealed the capping of the phytoconstituents, probably polyphenols from P. longum fruit extract and stabilizing the nanoparticles. Further the ferric ion reducing test, confirmed that the capping agents were condensed tannins. The aqueous P. longum fruit extract (PLFE) and the green synthesized silver nanoparticles (PLAgNPs) showed powerful antioxidant properties in in vitro antioxidant assays. The results from the antimicrobial assays suggested that green synthesized silver nanoparticles (PLAgNPs) were more potent against pathogenic bacteria than the P. longum fruit extract (PLFE) alone. The nanoparticles also showed potent cytotoxic effect against MCF-7 breast cancer cell lines with an IC 50 value of 67µg/ml/24h by the MTT assay. These results support the advantages of using bio-green method for synthesizing silver nanoparticles with antioxidant, antimicrobial and cytotoxic activities those are simple and cost effective as well.

HTLV 1 associated adult T cell lymphoma/leukemia a clinicopathologic, immunophenotypic tale of three cases from non-endemic region of south India.

Adult T cell lymphoma/leukemia is a peripheral T-cell neoplasm caused by human T-cell lymphotrophic virus-1, affects mostly adults with systemic involvement and poor prognosis. Diagnosis of adult T-Cell leukemia/Lymphoma is challenging. The clinico-pathologic and immuno-phenotypic features of the three cases will be presented.

CALLA negative precursor B lymphoblastic leukemia with MLL gene translocation and an unusual FISH signal pattern.

Rearrangements of the mixed lineage leukemia (MLL) gene at 11q23 commonly occur in infants with CALLA negative B lymphoblastic leukemia (B-ALL). Most often, these are detected by conventional karyotyping; however, fluorescent in-situ hybridization (FISH) with the help of a dual-color break-apart probe is used to identify cryptic translocations. When there is an MLL gene translocation, the usual FISH signal pattern is 1 red-1 green-1 yellow fusion signal pattern. We present a case of an infant with CALLA negative precursor B-ALL with a characteristic translocation t(4;11) (q21;q23), however, with an unusual MLL FISH signal pattern.

Undifferentiated sarcoma of the mitral valve with secondaries in brain in a girl of 22 years.

During a mitral valve replacement surgery in a girl of 22 years of age, it was accidentally discovered that the valve was destroyed due to a tumor and the histopathology and immunohistochemistry findings have proved it to be undifferentiated sarcoma. She was advised by the surgeon to go for chemotherapy. There was a delay of three months from the side of the patient to reach us and during that interval she has developed secondaries in the brain. This case is being presented here for its rarity.

Cloning and characterisation of the ahpA gene of Pasteurella multocida serogroup b:2 (strain P52): short communication.

Pasteurella multocida B:2 is responsible for haemorrhagic septicaemia in cattle and buffaloes, causing severe economic losses in the developing countries. In the present study, the ahpA gene of P. multocida B:2 (P52) was cloned, sequenced and compared with the previously reported ahpA gene sequence in P. multocida A:1, which is responsible for its haemolytic phenotype. E. coli DH5a cells were further transformed with recombinant plasmid carrying the ahpA gene from P. multocida B:2 (P52) but SDS-PAGE analysis failed to show the expression of haemolysin protein. Slight haemolysis was albeit observed in horse blood agar plates streaked with recombinant E. coli carrying the ahpA gene. Our study indicates that there is 99.6% similarity and 0.4% divergence between ahpA gene of P. multocida B:2 (P52) and P. multocida A: 1, while membrane topology analysis has predicted that ahpA is an inner membrane protein with two strong hydrophobic regions at the N and C terminals. The presence of significant homology in ahpA sequence in A: 1 and B:2 perhaps suggests a common mechanism of pathogenesis in different species of animals.