Primary signet ring cell carcinoma of prostate: A rare case report and review of literature.

Primary signet ring cell carcinoma (PSRCC) of the prostate is an extremely rare variant of prostatic adenocarcinoma. A PubMed search of the English language literature from January 2000 to June 2020 using the keywords "signet ring cell carcinoma" and "prostate," identified 20 cases of PSRCC of the prostate. On the basis of the combined data from this study and the literature review, 21 such patients were evaluated for clinical characteristics, histologic diagnoses, special and immunohistochemical staining, and treatment. The mean age at the diagnosis was 68.47 years (range 50-85 years). The prostate-specific antigen (PSA) levels varied from 0.19 to 6658 ng/mL, with a mean of 509.15 ng/mL. Most (50%) presented with Stage 3 cancer. The most common Gleason grade group was 5 (Gleason score 9 to 10), seen in 61.5%. The extent of signet ring cell involvement of the specimen when reported was documented as more than 20% of the tumor-containing signet ring cells, with a range of 25%-90%. For pathologic diagnosis, the most common special stains performed were periodic acid-Schiff and Alcian blue, and among the immunohistochemical stains, the most common were PSA, CK20, and prostate-specific acid phosphatase. A detailed clinicoradiological and pathological workup is essential to rule out primary from other common sites, in view of its grave prognosis and lack of an established treatment protocol.

An In-Vivo Comparative Study of the Soft Tissue Response and Esthetics of the Titanium Implant with Titanium Collar by Flapless and Conventional Flap Technique.

Background: These days, patients want quick results for tooth replacement and esthetic results. However, there is no direct correlation between the achievement of osseointegration and the outcome of successful treatment always. It is vital to sustaining peri-implant soft tissue health for extensive tenure success of the implant. Aim: The purpose of this trial was to determine, estimate, and compare the soft tissue retort and esthetics of the titanium implants with titanium collar at periodic intervals by flapless and conventional flap technique before and after prosthesis placement. Results: The difference in soft tissue indices namely, gingival index, plaque index, and the modified sulcular bleeding index was insignificant between the two implants placed by flapless and open flap technique 2. There was a significant difference amid the probing depths of the two implants after 3 months of prosthesis positioning where the implant placed by flapless technique showed lesser values as compared to the implant placed by the open flap technique 3. The esthetics of the soft tissues surrounding the titanium implant with titanium collar, when compared, presented a significant difference between the two techniques of implant placement. Conclusion: In conclusion, in recent advancements in dentistry, the flapless technique is becoming prominent because of procedure of minimally invasive surgery in implantology. There are advantages of early re-epithelialization and less inflammation around the soft tissue of the implant in the flapless procedure, provided that the prospective for the establishment of a fully functioning along with aesthetically desirable peri-implant soft tissue collar. The flapless technique accomplishes high degrees of gratification by the patients by shortening the surgery time and minimum invasion to both bone and soft tissue.

Biosynthesis of Silver Nanoparticles Using Azadirachta indica and Their Antioxidant and Anticancer Effects in Cell Lines.

In the present study, silver nanoparticles (Ag-NPs) were synthesized using Azadirachta indica extract and evaluated for their in vitro antioxidant activity and cytotoxicity efficacy against MCF-7 and HeLa cells. The silver nanoparticles (Ag-NPs) were formed within 40 min and after preliminary confirmation by UV-visible spectroscopy (peak observed at 375 nm), they were characterized using a transmission electron microscope (TEM) and dynamic light scattering (DLS). The TEM images showed the spherical shape of the biosynthesized Ag-NPs with particle sizes in the range of 10 to 60 nm, and compositional analysis was carried out. The cytotoxicity and antioxidant activity of various concentrations of biosynthesized silver nanoparticles, Azadirachta indica extract, and a standard ranging from 0.2 to 1.0 mg/mL were evaluated. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) activity of the biosynthesized Ag-NPs and aqueous leaf extract increased in a dose-dependent manner, with average IC50 values of the biosynthesized Ag-NPs, aqueous leaf extract, and ascorbic acid (standard) of 0.70 ± 0.07, 1.63 ± 0.09, and 0.25 ± 0.09 mg/mL, respectively. Furthermore, higher cytotoxicity was exhibited in both the MCF-7 and HeLa cell lines in a dose-dependent manner. The average IC50 values of the biosynthesized Ag-NPs, aqueous leaf extract, and cisplatin (standard) were 0.90 ± 0.07, 1.85 ± 0.01, and 0.56 ± 0.08 mg/mL, respectively, with MCF-7 cell lines and 0.85 ± 0.01, 1.76 ± 0.08, 0.45 ± 0.10 mg/mL, respectively, with HeLa cell lines. Hence, this study resulted in an efficient green reductant for producing silver nanoparticles that possess cytotoxicity and antioxidant activity against MCF-7 and HeLa cells.

Development and evaluation of a molecular based protocol for detection and quantification of Cryptosporidium spp. in wastewater.

Infections caused by protozoan parasites are a major public health concern globally. These infections are commonly diagnosed during water-borne outbreaks, necessitating accurate and highly sensitive detection procedures to assure public health protection. Current molecular techniques are challenged by several factors, such as low parasite concentration, inefficient DNA extraction methods, and inhibitors in environmental samples. This study focused on the development and validation of a molecular protocol for DNA extraction, efficient protozoan (oo)cyst recovery and quantification of protozoan parasites from wastewater using droplet digital polymerase chain reaction (ddPCR). Five DNA extraction methods, including commercial kits, custom phenol-chloroform, and in-house modified methods, were evaluated. The efficiency of each method was assessed via spectrophotometric analysis and ddPCR amplification using specific primers. Lastly, the developed protocol was evaluated for the detection and quantification of Cryptosporidium parvum in wastewater from different regions in South Africa. The conventional phenol-chloroform extraction method yielded the highest DNA concentration of 223 (±0.71) ng/µl and detected the highest number of Cryptosporidium parvum (1807 (±0.30) copies/ddPCR reaction) compared to other methods evaluated in this study. Additionally, the phenol-chloroform method demonstrated high sensitivity in extracting DNA from as few as one cyst/L of Cryptosporidium parvum, corresponding to 5.93 copies/ddPCR reaction. It was also observed that analysis of both the filtered supernatant and pellets after centrifugation improves the recovery efficiency of oocysts from wastewater by 10.5%, resulting in a total recovery of 64.1%. This optimized protocol was successfully applied to measure protozoan concentration in wastewater from different regions in South Africa. The improved DNA extraction and quantification method proposed in this study would be effective in monitoring protozoan concentration in the environment, which will help in instituting mitigation measures to reduce water-borne infections.

A review on application of next-generation sequencing methods for profiling of protozoan parasites in water: Current methodologies, challenges, and perspectives.

The advancement in metagenomic techniques has provided novel tools for profiling human parasites in environmental matrices, such as water and wastewater. However, application of metagenomic techniques for the profiling of protozoan parasites in environmental matrices is not commonly reported in the literature. The key factors leading to the less common use of metagenomics are the complexity and large eukaryotic genome, the prevalence of small parasite populations in environmental samples compared to bacteria, difficulties in extracting DNA from (oo)cysts, and limited reference databases for parasites. This calls for further research to develop optimized methods specifically looking at protozoan parasites in the environment. This study reviews the current workflow, methods and provide recommendations for the standardization of techniques. The article identifies and summarizes the key methods, advantages, and limitations associated with metagenomic analysis, like sample pre-processing, DNA extraction, sequencing approaches, and analysis methods. The study enhances the understanding and application of standardized protocols for profiling of protozoan parasite community from highly complexe samples and further creates a resourceful comparison among datasets without any biases.

Anterior segmental osteotomies without orthodontics: practicability of the correction of dentoalveolar deformities.

We designed this study to determine the efficiency and stability of anterior segmental osteotomies (ASO) without orthodontics for various dentofacial deformities. Records of patients treated with maxillary or mandibular ASO, or both, without orthodontics in the past 15 years were analysed. The assessment included postoperative analysis of patients' aesthetics and functional satisfaction using a questionnaire and grading (score 0 - 4) system, and the amount of relapse calculated from 12-month postoperative cephalograms. A total of 26 ASO subjects (age range 13- 31 years) were studied (14 maxillary, two mandibular, and 10 bimaxillary). Long-term stability was acceptable in all cases with no significant relapse (p>0.05). No major complications were encountered. All patients reported good to excellent (score=3 to 4) satisfaction following surgery. Using meticulous planning and a careful surgical technique, ASO without orthodontics is a simple, quick, safe, and stable option for the correction of dentofacial deformities.

Anesthetic challenges while performing emergency laparotomy in a patient having COVID-19 infection.

An elderly hypertensive lady presented with fever, respiratory symptoms, and mild abdominal discomfort and was diagnosed to have COVID-19 pneumonia. Respiratory symptoms improved with steroids, awake proning, high flow nasal cannula oxygen therapy and antibiotics. After 4 days, she developed non-occlusive superior mesenteric artery thrombosis, which initially responded to anticoagulants but was complicated on tenth day by intestinal obstruction necessitating emergency surgery. Challenges encountered perioperatively were multi systemic involvement, pneumonia, ventilation- perfusion mismatch, sepsis along with technical difficulties like fogging of goggles, stuck expiratory valve on anesthesia machine, inaudibility through stethoscope and discomfort due to personal protective equipment. Perioperative focus should be on infection prevention, maintenance of hemodynamics, and optimization of oxygenation with preoperative high flow nasal cannula oxygen therapy. Ultrasound lung helps in correct placement of endotracheal tube. We recommend daily machine check, taping of N95 mask to face and ambient operation theatre temperatures of 20-22°C to reduce technical problems.

Expression of p27 and p16 and their clinical significance in gastric cancer.

BACKGROUND: Deregulated expression of cell cycle regulators p27 and p16 is associated with cancer progression. p27kip1 and p16INKa are a cyclin dependent kinase inhibitor whose major target is the cyclinE/CDK2 and cyclinD/CDK4/6 complex, respectively, that governs cell cycle transition from late G1 to S phase. METHODS: We recruited biopsies of a total of 84 subjects including 72 primary tumor biopsies from histopathologically proven gastric carcinoma, 8 adjacent controls and 12 independent controls. We used gastric cancer cell line, AGS, for validation of our data. Expression profiling at transcript level was done by semi-quantitative RT-PCR and at proteome level by immunohistochemistry and immunofluorescence. Receiver operator characteristics analysis was done for determining the diagnostic utility of p27 and p16 with respect to the sensitivity and specificity. RESULTS: We demonstrate that p27 and p16 are frequently over expressed in early stages of gastric carcinoma. Our semi-quantitative data show a significant upregulation of p27 (Mean ± SEM, 0.4771 ± 0.0895; p = 0.0001) and p16 (Mean ± SEM, 0.4676 ± 0.04305; p = 0.0001) at mRNA level. Concordant to semi-quantitative data, immunohistochemistry data also showed a significant upregulation of p27 (Mean ± SEM, 196.4 ± 10.84; p < 0.0001) and p16 (Mean ± SEM, 100.4 ± 23.71; p < 0.0001) at protein level. CONCLUSIONS: The present study showed that the significant upregulation of p27 and p16 were associated with early events in gastric carcinogenesis. Our data suggests that clinical correlation of these differentially expressed genes may be useful as diagnostic biomarkers for early detection of gastric carcinoma and promising therapeutics target for GC patients.

Association of UGT1A6 gene polymorphism with clinical outcome in pediatric epileptic patients on sodium valproate monotherapy

Pediatric epilepsy comprises chronic neurological disorders characterized by recurrent seizures. Sodium valproate is one of the common antiseizure medications used for treatment. Glucuronide conjugation is the major metabolic pathway of sodium valproate, carried out by the enzyme uridine 5′-diphosphate (UDP) glucuronosyl transferase (UGT) whose gene polymorphisms may alter the clinical outcome. The objective of this study was to assess the association between UGT1A6 genetic polymorphism and clinical outcome in terms of efficacy and tolerability in pediatric epileptic patients on sodium valproate monotherapy. Pediatric epileptic patients (n=65) aged 2-18 years receiving sodium valproate monotherapy for the past one month were included. Genetic polymorphism patterns of UGT1A6 (T19G, A541G, A552C) were evaluated by PCR-RFLP. Clinical outcome was seizure control during the 6 months observation period. Tolerability was measured by estimating the hepatic, renal, and other lab parameters. Out of 65 patients, TT (40%), TG (57%), and GG (3%) patterns were observed in UGT1A6 (T19G) gene, AA (51%), AG (40%), and GG (9%) in (A541G) gene, and AA (43%), AC (43%), and CC (14%) in (A552C) gene. No statistical difference in clinical outcome was found for different UGT1A6 genetic polymorphism patterns. We concluded that different patterns of UGT1A6 genetic polymorphism were not associated with the clinical outcome of sodium valproate in terms of efficacy and tolerability. Sodium valproate was well-tolerated among pediatric patients with epilepsy and can be used as an effective antiseizure medication.

A comparison of poly-ethylene-glycol-coated and uncoated gold nanoparticle-mediated hepatotoxicity and oxidative stress in Sprague Dawley rats.

BACKGROUND: Gold nanoparticles (GNPs) and their functional derivatives are of great interest because of their many biomedical applications. GNPs are increasingly being incorporated into new diagnostic and therapeutic approaches in medicine. Consequently, there has been a strong push to fully understand their interactions with blood components. The agglomeration of cells reflects the interaction of nanoparticles with blood components. METHODS: The main aim of this study was to compare the effects of poly-ethylene-glycol (PEG)-oated and uncoated GNPs on the generation of reactive oxygen species (ROS); on the actions of distinct hepatotoxicity biomarkers such as alanine (ALT) and aspartate (AST) aminotransferases, and alkaline phosphatase (ALP); and on the histology of liver tissues in the rat model. Four distinct doses of PEG-coated and uncoated GNPs (12.5, 25, 50, and 100 µg/kg body weight) were used. Each group consisted of three rats receiving an oral administration of PEG-coated and uncoated GNPs for 5 days with one dose per 24 hours. The control group consisted of three rats that received deionized water. Twenty-four hours after the last treatment, samples were collected following standard procedures. RESULTS: PEG-coated and uncoated GNPs enhanced the generation of ROS and the activity of serum aminotransferases (ALT/AST) and ALPs relative to the negative control. A liver histology assessment of GNP-exposed rats revealed statistically significant responses in the variation of the morphologies of tissues relative to those of the negative control. Nonetheless, uncoated GNPs demonstrated enhanced hepatotoxic outcomes relative to those of PEG-coated GNPs. The results demonstrated that both GNPs may be able to promote hepatotoxicity in Sprague Dawley rats through mechanisms of oxidative stress. However, uncoated GNPs have more harmful effects than PEG-coated GNPs relative to the negative control. CONCLUSION: Taken together, the results of this study indicate that PEG-coated GNPs may be safer to use in nanomedicinal applications than uncoated GNPs. However, more studies must be performed to confirm the outcomes of PEGylation.

Utility of a Single-Tube, Six-Color Flow Cytometry Panel for the Diagnosis of Myelodysplastic Syndrome: Experience of a Tertiary Care Centre in India.

Background: Diagnosis of myelodysplastic syndromes (MDS) is challenging in the presence of morphological mimickers. Flow cytometric immunophenotyping (FCI) has been added to the diagnostic armamentarium, but its use in clinical practice is variable. Materials and Methods: Bone marrow aspirate samples from 54 patients with a clinical and/or morphological suspicion of MDS were subjected to FCI using a single-tube, 6-colour panel comprising of monoclonal antibodies against CD13, CD11b, CD16, CD34, CD45 and CD56. Analysis was centered on the abnormal maturation pattern of granulocytes, blast percentage (≥3%) and ratio of side scatter peak channel value (SSC-PCV) of granulocytes and lymphocytes. Each of these parameters was assigned a score of 1. Overall sensitivity and specificity of this panel was ascertained to differentiate cytopenia/s of MDS from non-MDS cases. Results: Forty MDS and 14 non-MDS cases were diagnosed based on morphology and cytogenetic results. Twenty control samples were also processed simultaneously for FCI to assign the cutoff for various flow cytometric parameters. A score ≥2 was defined as positive for MDS. Hypogranularity was present in 62.5% cases of MDS. The median SSC-PCV of granulocytes and lymphocytes was 6.16 in the MDS group, 7.9 in the non-MDS group and 8.90 in the control group (p <0.05). This cut-off value of 6.16 had a specificity of 92.5% based on the ROC curve analysis. Abnormal granulocyte maturation patterns for CD13/16, CD13/11b and CD11b/16 dot plots were observed in 95.3, 69.8 and 74.4% cases, respectively. The overall sensitivity and specificity of the panel was found to be 87.5% and 64.2%, respectively. Conclusion: FCI is now an important tool for diagnostic workup in patients presenting with persistent cytopenia with or without morphological evidence of dyspoiesis. Inclusion of objective parameters like SSC-PCV would also reduce inter-lab variability in MDS diagnosis.

Sulfamic acid promoted one-pot synthesis of phenanthrene fused-dihydrodibenzo-quinolinones: Anticancer activity, tubulin polymerization inhibition and apoptosis inducing studies.

A facile one-pot method for the synthesis of new phenanthrene fused-dihydrodibenzo-quinolinone derivatives has been successfully accomplished by employing sulfamic acid as catalyst. These new compounds were evaluated for their in vitro cytotoxic potential against human lung (A549), prostate (PC-3 and DU145), breast (MCF-7) and colon (HT-29 and HCT-116) cancer cell lines. Among all the tested compounds, one of the derivatives 8p showed good anti-proliferative activity against A549 lung cancer cell line with an IC50 of 3.17 ±â€¯0.52 µM. Flow cytometric analyses revealed that compound 8p arrested both Sub G1 and G2/M phases of cell cycle in a dose dependent manner. The compound 8p also displayed significant inhibition of tubulin polymerization and disruption of microtubule network (IC50 of 5.15 ±â€¯0.15 µM). Molecular docking studies revealed that compound 8p efficiently interacted with critical amino acid Cys241 of the α/ß-tubulin by a hydrogen bond (SH…O = 2.4 Å). Further, the effect of 8p on cell viability was also studied by AO/EB, DCFDA and DAPI staining. The apoptotic characteristic features revealed that 8p inhibited cell proliferation effectively through apoptosis by inducing the ROS generation. Analysis of mitochondrial membrane potential through JC-1 staining and annexin V binding assay indicated the extent of apoptosis in A549 cancer cells.

Does platelet-rich fibrin have a role in osseointegration of immediate implants? A randomized, single-blind, controlled clinical trial.

Immediate implants are a valuable treatment option to replace natural teeth in the aesthetic region. The hypothesis of this randomized controlled clinical trial was that immediate implants grafted with autologous platelet-rich fibrin (PRF) have better clinical and radiographic outcomes than non-grafted controls. Forty-one implants were placed in 31 subjects with one or more non-restorable single-rooted teeth. Autologous PRF was placed in the peri-implant region of the study group (n=21) and no augmentation was done in the control group (n=20). A staged protocol was followed for implant restoration. The patients received a definitive restoration after 3 months and were followed up for a period of 1year. The statistical analysis included 39 implants sites in 29 subjects. A significant increase in implant stability was noted in both groups over the 3-month period (implant stability quotient: study group 56.58±18.81 to 71.32±7.82; control group 60.61±11.49 to 70.06±8.96; P=0.01). No significant difference was observed between the groups in terms of implant stability. The hypothesis was thus rejected, as there was no significant effect of PRF on immediate implants with adequate primary stability.

Porcine sapelovirus among diarrhoeic piglets in India.

Porcine sapelovirus (PSV) A belongs to the genus Sapelovirus, family Picornaviridae. PSV infections in pigs have been reported from European countries, United States, Japan, China, Korea and Brazil. The virus has been isolated/detected from faeces of healthy pigs as well as those affected with diarrhoea, respiratory signs, encephalitis, skin lesions and fertility disorders. This study was planned to investigate whether PSV is prevalent among pigs in India and to characterize PSV encountered in the study population. The study revealed that five of 70 (7.14%) faecal samples were found positive for PSV using RT-PCR. Three viruses were successfully isolated from faecal samples using IB-RS-2 cell line. Complete genome sequencing and analysis of one Indian PSV isolate revealed highest homology (88%) with V13 strain from England. Phylogenetic analysis of the complete polyprotein nucleotide sequences of 14 strains of PSV classified the viruses into four distinct clades. This first report from India adds to our knowledge on genetic diversity of PSV detected so far among pigs in different countries. A large-scale surveillance of the virus is required to understand its genomic diversity and economic impact.

Synthesis of substituted phenanthrene-9-benzimidazole conjugates: Cytotoxicity evaluation and apoptosis inducing studies.

A series of new phenanthrene-9-benzimidazole conjugates has been synthesized by condensing phenanthrene aldehydes with various substituted o-phenylenediamines. The title compounds were evaluated for their in vitro cytotoxic potential against various human cancer cell lines like breast (BT-549), prostate (PC-3 and DU145), triple negative breast cancer (MDA-MB-453), and human colon cancer (HCT-116 and HCT-15) cells. Among the tested compounds, 10o displayed significant in vitro cytotoxic activity against PC-3 prostate cancer cells with an IC50 value of 6.32 ± 0.09 µM. Further, the cell cycle analysis indicated that it blocks G2/M phase of the cell cycle in a dose dependent manner. In order to determine the effect of the compound 10o on cell viability; phase contrast microscopy, AO/EB staining, DAPI staining, and DCFDA staining studies were performed. In these studies, apoptotic features were clearly observed indicating that the compound inhibited cell proliferation by apoptosis. JC-1 staining and annexin binding assays indicated the extent of apoptosis in PC-3 cells. Further, relative viscosity measurements and molecular docking studies indicated that these compounds bind to DNA by intercalation.