[Conventional paraffin embedding technique and double-embedding technique for minute oral biopsies and delicate pulp tissue - a comparative study]. / Traditsionnyi metod zalivki v parafin i metod dvoinoi zalivki dlya malen'kikh bioptatov polosti rta i myagkikh tkanei pul'py ­ sravnitel'noe issledovanie.

BACKGROUND: Among oral biopsies, small incisional tissues, have to be preserved all through the processing and embedding to ensure optimal visualization of all the mucosal layers without compromise. Optimal tissue orientation is the most critical step in tissue processing for demonstration of definitive morphology in the sections, which is often more challenging in cases of minute/small or thinner sections using routine paraffin techniques to evaluate accurate diagnosis. Some modification is needed to handle these samples to get a better result. Double embedding technique with some modification has been widely used for small/ thin/ multiple biopsies and gives excellent results in many other fields like general pathology and biotechnology. The double embedding technique though produced excellent and significant results in mucosal biopsies yet, it is of minimal interest among oral pathologists. To best of our knowledge, this is the first study to use double embedding technique for pulp tissues. OBJECTIVE: The present study was aimed to evaluate and compare the ease of embedding and sectioning sections using Agar-Paraffin double embedding technique for small oral mucosal biopsies and thin pulp tissues. MATERIAL AND METHODS: A total of 40 oral tissue samples categorized into two groups were taken for the present study. Group I included 20 small oral mucosal biopsy samples of size ranging from 0.2 to 0.5 cm and Group II included 20 pulp tissues obtained from freshly extracted non carious tooth. 10 blocks were prepared by routine paraffin method and 10 blocks were prepared by modified double embedding method for each group. Scores were given by comparing all the criteria with that of the routine paraffin technique. Chi-square test was used for statistical analysis. RESULTS: The average ease score for the Agar-Paraffin double embedded small/minute biopsies showed better scores than the pulp tissue with that of the routine technique. However, no statistically significant difference was seen among embedding and sectioning sections between the two groups. CONCLUSION: Modified double embedding method is simple and reliable alternative technique that helps in better orientation, processing and sectioning especially for oral small or thin biopsies and delicate pulp tissues.

Dysplasia severity is associated with poor quality of life in patients with Barrett's esophagus referred for endoscopic eradication therapy.

Limited data exist regarding patient-reported outcomes and quality of life (QOL) experienced by patients with Barrett's esophagus (BE) referred for endoscopic eradication therapy (EET). Specifically, the impact of grade of dysplasia has not been explored. The purpose of this study is to measure patient-reported symptoms and QOL and identify factors associated with poor QOL in BE patients referred for EET. This was a prospective multicenter study conducted from January 2015 to October 2017, which included patients with BE referred for EET. Participants completed a set of validated questionnaires to measure QOL, symptom severity, and psychosocial factors. The primary outcome was poor QOL defined by a PROMIS score >12. Multivariable logistic regression analysis was performed to identify factors associated with poor QOL. In total, 193 patients participated (mean age 64.6 years, BE length 5.5 cm, 82% males, 92% Caucasians) with poor QOL reported in 104 (53.9%) participants. On univariate analysis, patients with poor QOL had lower use of twice daily proton pump inhibitor use (61.5% vs. 86.5%, P = 0.03), shorter disease duration (4.9 vs. 5.9 years, P = 0.04) and progressive increase in grade of dysplasia (high-grade dysplasia: 68.8% vs. 31.3%, esophageal adenocarcinoma: 75.5% vs. 24.5%, P < 0.001). Multivariate analysis demonstrated that high-grade dysplasia was independently associated with poor QOL (OR: 5.57, 95% CI: 1.05, 29.5, P = 0.04). In summary, poor QOL is experienced by the majority of patients with BE referred for EET and the degree of dysplasia was independently associated with poor QOL, which emphasizes the need to incorporate patient-centered outcomes when studying treatment of BE-related dysplasia.

Effect of lambda cyhalothrin and temephos on detoxification enzyme systems in Culex quinquefasciatus (Diptera: Culicidae).

Mosquitoes serve as vector for transmitting diseases. Among mosquitoes, Culex quinquefasciatus transmits lymphatic filariasis, yellow fever Japanese encephalitis etc. Application of chemical insecticides is still the best option for vector control programmes. Continuous use of these chemicals on mosquito reduces its effects. The present study determined the baseline susceptibility of Cx. quinquefasciatus in response to λ-cyhalothrin and temephos treatments. In addition, the biochemical mechanisms and zymogram analysis involved in insecticide detoxification among larval mosquitoes were studied. The larval bioassay indicated high LC50 value for λ-cyhalothrin (0.1484ppm) as compared to temephos (0.01092ppm). While AChE assay showed increased activity in temephos treatments, glutathione reductase (GR) and esterase levels were increased at both the treatments. Esterase quantitative analysis revealed the expression of three bands at 43kDa, 67kDa and 245kDa. The findings suggest that insensitivity of AChE, esterase and high GR activity may play an important role in developing resistance to synthetic pyrethroid and organophosphate insecticides in Cx. quinquefasciatus population.

Susceptibility status of Aedes aegypti (L.) (Diptera: Culicidae) to temephos from three districts of Tamil Nadu, India.

BACKGROUND & OBJECTIVES: Dengue is the most rapidly expanding arboviral disease in India. Aedes aegypti is the primary vector of dengue fever. Chemical insecticides have long been used in the vector control programmes along with other control measures. However, continuous use of insecticides targeting Ae. aegypti may lead to development of insecticide resistance. Though resistance in Ae. aegypti has been reported, the mutation in ace-1 gene associated with temephos resistance is not reported in natural populations. The present study aims to evaluate the susceptibility/resistance status of Ae. aegypti to temephos from three districts of Tamil Nadu. METHODS: Ae. aegypti larvae were sampled from different locations in three districts, viz., Dharmapuri, Salem and Namakkal. The standard WHO larval bioassay, biochemical assays and spotting of specific mutation (G119S) in the acetylcholinesterase gene, which is associated with organophosphate resistance, were carried out by PCR and sequencing. RESULTS: The results showed that larvae from Namakkal (NKL) population had an alteration in their susceptibility status (RR = 6.9 fold), while the other populations were moderately susceptible to insecticides. Biochemical assay showed increased activity for α- and ß-esterase in NKL, as well as evidence of acetylcholinesterase insensitivity. G119S mutation was detected in this population with high frequency of 0.24. INTERPRETATION & CONCLUSION: The high activity of esterase, mixed-function oxidase (MFO) and ace-1 mutation frequency were closely associated with temephos resistance. Early detection of resistance alleles in natural vector population could be useful for the successful implementation of insecticide resistance management strategies. The results of this study provide baseline data on temephos resistance in Ae. aegypti populations.

Involvement of metabolic resistance and F1534C kdr mutation in the pyrethroid resistance mechanisms of Aedes aegypti in India.

Pesticide resistance poses a serious problem for worldwide mosquito control programs. Resistance to insecticides can be caused by an increased metabolic detoxification of the insecticide and/or by target site insensitivity. In the present study, we estimated the tolerance of Indian Aedes aegypti populations using adult bioassays that revealed high resistance levels of the field populations to permethrin (RR-6, 5.8 and 5.1 folds) compared to our susceptible population. Enzymatic assays revealed increased activities of glutathione S-transferase and carboxylesterase enzymes in the field populations comparatively to the susceptible population. PBO synergist assays did not confirm that cytochrome P450 monooxygenase metabolic detoxification acted as a major cause of resistance. Hence the role of target site resistance was therefore investigated. A single substitution Phe1534Cys in the voltage gated sodium channel was found in domain III, segment 6 (III-S6) of the resistance populations (allele frequency=0.59, 0.51 and 0.47) suggesting its potential role in permethrin resistance in A. aegypti.

Resistance selection and molecular mechanisms of cypermethrin resistance in red hairy caterpillar (Amsacta albistriga walker).

Amsacta albistriga is one of the important pests of oilseed crops in India. This pest has developed high resistance to organophosphate (OP) insecticide in field. Therefore, cypermethrin insecticide was used as an alternative for this pest. After 20 generations of selection with cypermethrin, the LD50 value for A. albistriga was increased by 21.5-folds. The synergism ratio of piperonyl butoxide (PBO) and triphenyl phosphate (TPP) was increased by 10- and 9.6-fold in resistant strains and comparatively, 3.9 and 4.2-fold in susceptible strains. Detoxification enzyme analysis and native PAGE electrophoresis of esterase isoenzyme further revealed that esterase and mixed function oxidase may be involved in cypermethrin resistance in CypRes strain. In addition to enzyme analysis overexpression of CYP4M44, CYP9A77 and CYP6B47 (ortholog) can confer metabolic resistance in the CypRes strain. These data provide a foundation for further study of cypermethrin resistance mechanism observed in A. albistriga.

Prevalence of Simkania negevensis in chlorinated water from spa swimming pools and domestic supplies.

AIMS: This study aimed to investigate the prevalence of Simkania negevensis in the chlorinated water of spa swimming pools and domestic network systems. METHODS AND RESULTS: A total of 10 and 36 samples were collected from two domestic water supplies and four spa facilities, respectively. Simkania negevensis was isolated in Acanthamoeba castellanii monolayers and detected by Gimenez staining and immunofluorescence test. Simkania negevensis DNA, extracted from the positive samples, was confirmed by a nested PCR assay followed by sequencing. Simkania negevensis was detected in 41·3% of samples (domestic water: 50%; untreated spa supply water: 25%; chlorinated spa pool water: 42·9%) from all the examined water systems in successive samplings performed in 1 year. The presence of S. negevensis was not correlated with the counts of Heterotrophic Bacteria and Pseudomonas aeruginosa. Unlike Legionella spp., which were never isolated from the swimming-pool water samples, S. negevensis was also detected in chlorinated spa water. CONCLUSIONS: This investigation shows that Simkania is widespread in natural and man-made aquatic environments, which may represent possible sources of infection: in the swimming pools, in particular, the aerosol generated by the water movement could increase the risk of inhalation of infected particles. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first evidence of the presence of S. negevensis in spa pool water. Sero-epidemiological surveys on spa users could help to clarify its transmission in this environment.

ST elevation in ECG lead aVR signals severe acute left main coronary artery disease.

Patients with suspected acute coronary syndrome are commonly assessed by acute physicians on arrival in hospital. Although most will recognise the typical ECG features of ST elevation myocardial infarction, the significance of ST elevation in lead aVR may not always be appreciated. This case series describes 6 cases in which this ECG abnormality was the predominant feature in patients whose subsequent angiogram revealed severe acute left main coronary artery disease. The importance of early referral of such patients to a centre in which percutaneous coronary intervention can be performed, is discussed.

Long-term benefits of testosterone replacement therapy on angina threshold and atheroma in men.

INTRODUCTION: In short-term studies, testosterone replacement therapy has been shown to protect male subjects from exercise-induced ischaemia and modify cardiovascular risk factors such as insulin resistance, fat mass and lipid profiles. METHODS: This randomised parallel group controlled trial was designed to assess the treatment effect of testosterone therapy (Nebido) compared with placebo in terms of exercise-induced ischaemia, lipid profiles, carotid intima-media thickness (CIMT) and body composition during 12 months treatment in men with low testosterone levels and angina. RESULTS: A total of 15 men were recruited but 13 (n=13) reached adequate duration of follow-up; seven were treated with testosterone and six with placebo. Testosterone increased time to ischaemia (129+/-48 s versus 12+/-18, P=0.02) and haemoglobin (0.4+/-0.6 g/dl versus -0.03+/-0.5, P=0.04), and reduced body mass index (-0.3 kg/m(2) versus 1.3+/-1, P=0.04) and triglycerides (-0.36+/-0.4 mmol/l versus 0.3+/-1.2, P=0.05). The CIMT decreased in the testosterone group more than placebo, but full between group analyses suggested this was only a statistical trend (-0.5+/-0.1 vs -0.09+/-0.06, P=0.16). There were no significant effects on serum prostate specific antigen, total or high-density lipoprotein cholesterol; or on mood and symptom scores as assessed by Seattle Angina Score and EuroQol. CONCLUSION: The protective effect of testosterone on myocardial ischaemia is maintained throughout treatment without decrement. Previously noted potentially beneficial effects of testosterone on body composition were confirmed and there were no adverse effects.

Endoscopic ablation of intestinal metaplasia containing high-grade dysplasia in esophagectomy patients using a balloon-based ablation system.

BACKGROUND: This study aimed to determine the optimal treatment parameters for the ablation of intestinal metaplasia (IM) containing high-grade dysplasia (HGD) using a balloon-based ablation system for patients undergoing esophagectomy. METHODS: Immediately before esophagectomy, patients underwent ablation of circumferential segments of the esophagus containing IM-HGD using the HALO360 system. The treatment settings were randomized to 10, 12, or 14 J/cm2 for two, three, or four applications. After esophagectomy, multiple sections from ablation zones were microscopically evaluated. Histologic end points included maximum ablation depth (histologic layer) and complete ablation of all IM-HGD (yes/no). RESULTS: Eight men with a mean age of 57 years (range, 45-71 years) were treated, and 10 treatment zones were created. There were no device-related adverse events. At resection, there was no evidence of a transmural thermal effect. Grossly, ablation zones were clearly demarcated sections of ablated epithelium. The maximum ablation depth was the lamina propria or muscularis mucosae. The highest energy (14 J/cm2, 4 applications) incurred edema in the superficial submucosa, but no submucosa ablation. Complete ablation of IM and HGD occurred in 9 of 10 ablation zones (90%), defined as complete removal of the epithelium with only small foci of "ghost cells" representing nonviable, ablated IM-HGD and demonstrating loss of nuclei and cytoarchitectural derangement. One focal area of viable IM-HGD remained at the margin of one ablation zone (12 J/cm2, 2 applications) because of incomplete overlap. CONCLUSION: Complete ablation of IM-HGD without ablation of submucosa is possible using the HALO360 system. Ablation depth is dose related and limited to the muscularis mucosae. In one patient, small residual foci of IM-HGD at the edge of the ablation zone were attributable to incomplete overlap, which can be avoided. This study, together with nonesophagectomy IM-HGD trials currently underway, will identify the optimal treatment parameters for IM-HGD patients who would otherwise undergo esophagectomy or photodynamic therapy.

Thoracoscopic resection of esophageal duplication cysts.

Esophageal duplication is a rare congenital esophageal disorder. Surgical excision is the standard treatment for symptomatic esophageal duplication cysts. Traditionally, the resection is accomplished via thoracotomy; however, a minimally invasive approach is possible, avoiding the long hospital stay, the discomfort and the long recovery time due to a thoracotomy. The authors describe two cases of esophageal duplication resected via a left thoracoscopic approach.

Active inflammatory bowel disease and coronary artery dissection.

The case of a young woman with flare-up of Crohn's disease, who had an acute myocardial infarction due to the spontaneous dissection of the left anterior descending coronary artery, is reported. The literature on this rare condition is reviewed and a mechanism postulated for spontaneous coronary artery dissection in inflammatory bowel disease.

Role of human amniotic epithelial cell transplantation in spinal cord injury repair research.

Human amniotic epithelial cells (HAEC) possess certain properties similar to that of neural and glial cells. In the present work, the potential of HAEC as stem cells for spinal cord injury repair was tested. HAEC obtained from human placenta were labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyllindocarbocyanine perchlorate (Dil) in the culture medium. These labeled cells were transplanted into the transection cavities in the spinal cord of bonnet monkeys. Results were analyzed after 15 and 60 days of post-transplantation. HAEC cells survived in the monkey spinal cord for up to the maximum period of observation in the present study, i.e. 60 days. HAEC graft was penetrated by the host axons. There was no glial scar at the transection lesion site. Some of the host spinal neurons and axons were labeled with Dil (used to label HAEC) whereas in lesion control group, there was no such host-neuron labeling. This may be either due to the prevention of death in the axotomized neuron's ensuing lesion or due to the neurotrophic effect exhibited by the transplanted HAEC. Further studies would be required to verify these speculations. Therefore from this pilot study it appears that HAEC survive in the transplanted environment, support the growth of host axons through them, prevent the formation of glial scar at the cut ends and may prevent death in axotomized cells or attract the growth of new collateral sprouting. The abovementioned properties, i.e. serving as a suitable milieu for the host axons to grow, preventing glial scar at the lesion site and rescuing axotomized neurons from death were previously reported in the case of neural transplantation studies. Thus it is speculated that HAEC may be having certain properties equal to the beneficial effects of neural tissue in repairing spinal cord injury. Apart from this speculation, there are two more reasons for why HAEC transplantation studies are warranted to understand the long-term effects of such transplantations. First, there was no evidence of immunological rejection probably due to the non-antigenic nature of the HAEC. Second, unlike neural tissue, procurement of HAEC does not involve many legal or ethical problems.