Hematological and plasma biochemistry in Cirrhinus mrigala (Hamilton 1822)

Hematological and plasma chemistry indices are important parameters for the evaluation of fish physiological status. In this study, we determined the results of selected hematological and plasma biochemical profiles in Cirrhinus mrigala captured from the wild in a tropical climate of India. Blood was analyzed using standard techniques, and differences in hematological parameters including hemoglobin concentration, erythrocyte count, total leukocyte count, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) of fish were compared according to sex and different seasons. Analysis of variance showed that there were significant differences (p?<?0.05) between sexes, and the results indicated that blood parameters levels between the sexes in summer were significantly different than that measure in other seasons except MCH and MCHC value. The number of total leukocyte levels was found to be higher in female fish especially in reproductive seasons (summer), but the levels of hemoglobin, hematocrit, and MCV values were high in male fish in an annual period. However, there was no difference in MCH and MCHC values between the sexes and seasons throughout the study period. These may be related to season of sampling and changing physiological cycles during these months. Differences were noted in plasma lipid, cholesterol, and glucose level in the summer season, but the higher amount of plasma protein was found in spring and winter in male and female fish, respectively. In conclusion, monitoring fish hematological and biochemical parameters essentially can be a way to evaluate the physiological and health status of their populations, which may be a useful indicator of the environmental status.     

Evaluation of Vertical Bone Gain Following Alveolar Distraction Osteogenesis in the Anterior Edentulous Mandible: A Clinical Study

Aim and Objectives

To calculate the gain in bony height and volume of the distracted upper segment using an extraosseous unidirectional device to improve the retention of the future prosthesis.

Materials and Methods

Ten cases with completely or partially edentulous but severely or considerably resorbed anterior mandibles were managed by vertical alveolar distraction osteogenesis. All the patients were evaluated preoperatively, intraoperatively and post-operatively for various parameters clinically and radiographically i.e. on OPG, lateral cephalogram and on CT scan, at different time intervals. In all cases there was increase in vertical bone height.

Results

The study showed mean vertical bone gain (VBG) on OPG as 8.2 mm. The mean VBG on lateral cephalogram was 8.1 mm. The mean VBG on CT scan at right canine was 8.35 mm, at left canine was 8.2 mm and at midline was 8.27 mm.

Conclusion

Alveolar distraction osteogenesis is a predictable method for restoring alveolar ridges prior to implant placement or prosthesis. Distraction osteogenesis is ideally suited for recreating missing tissue in the anterior esthetic zone by increasing vertical bone height as well as good width and soft tissue growth.     

Comparison of bronchoscopic and non-bronchoscopic techniques for diagnosis of ventilator associated pneumonia

Background:

The diagnosis of ventilator associated pneumonia (VAP) remains a challenge because the clinical signs and symptoms lack both sensitivity and specificity and the selection of microbiologic diagnostic procedure is still a matter of debate.

Aims and Objective:

To study the role of various bronchoscopic and non-bronchoscopic diagnostic techniques for diagnosis of VAP.

Settings and Design:

This prospective comparative study was conducted in a medical ICU of a tertiary care center.

Materials and Methods:

Twenty-five patients, clinically diagnosed with VAP, were evaluated by bronchoscopic and non-bronchoscopic procedures for diagnosis. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of various bronchoscopic and non-bronchoscopic techniques were calculated, taking clinical pulmonary infection score (CPIS) of ≥6 as reference standard.

Results:

Our study has shown that for the diagnosis of VAP, bronchoscopic brush had a sensitivity, specificity, PPV and NPV of 94.9% [confidence interval (CI): 70.6–99.7], 57.1% (CI: 13.4–86.1), 85% (CI: 61.1–96) and 80% (CI: 21.9–98.7), respectively. Bronchoscopic bronchoalveolar lavage (BAL) had a sensitivity, specificity, PPV and NPV of 77.8% (CI: 51.9–92.6), 71.8% (CI: 24.1–94), 87.3% (CI: 60.4–97.8) and 55.5% (CI: 17.4–82.6), respectively. Sensitivity, specificity, PPV and NPV for non–bronchoscopic BAL (NBAL) were 83.3% (CI: 57.7–95.6), 71.43% (CI: 24.1–94), 88.2% (CI: 62.3–97.4) and 62.5% (CI: 20.2–88.2), respectively. Endotracheal aspirate (ETA) yield was only 52% and showed poor concordance with BAL (κ-0.351; P-0.064) and NBAL (k-0.272; P-0.161). There was a good microbiologic concordance among different bronchoscopic and non-bronchoscopic distal airway sampling techniques.

Conclusion:

NBAL is an inexpensive, easy, and useful technique for microbiologic diagnosis of VAP. Our findings, if verified, might simplify the approach for the diagnosis of VAP.     

Ras gene product expression in blood and marrow smears of patients with acute leukemia: importance of fixation

Activation of ras protooncogenes by any of several possible mutations in codons 12, 13 or 61 has been demonstrated in a variety of human malignancies, including acute non-lymphoblastic leukemia (ANLL). In situ staining for the ras gene product, p21, has been demonstrated in carcinomas of several sites. High levels of p21 expression have been associated with histologic anaplasia in prostate cancer and regional lymph node metastasis in breast cancer. We examined 16 marrow aspirates and blood smears from patients with acute leukemia, predominantly ANLL, and eight controls. Marrow aspirates or blood were smeared on glass slides and fixed immediately in 10% buffered formalin. p21 was examined with avidin-biotin linked immunoperoxidase visualization. Particular attention must be paid to antibody selection and fixation protocol to demonstrate p21, owing to its rapid degradation ex vivo. Three of 16 patients exhibited occasional high p21 expression primarily in leukemic blasts, but in no case were more than 10% of blast cells positive. Normal reticuloendothelial and myeloid cells occasionally exhibited mild to moderately heavy staining, but megakaryocytes, erythroid precursors, lymphocytes and plasma cells were consistently negative. Most patients, 5 normal volunteers and 3 patients with non-malignant disease, exhibited no reactivity, or only a faint blush. These data suggest that while point mutation and concomitant activation of c-N-ras occurs regularly in ANLL, high levels of ras p21 expression are rarely found with this technique.     

Selection of antigens and development of prototype tests for point-of-care leprosy diagnosis

Leprosy can be a devastating chronic infection that causes nerve function impairment and associated disfigurement. Despite the recent reduction in the number of registered worldwide leprosy cases as a result of the widespread use of multidrug therapy, the number of new cases detected each year remains relatively stable. The diagnosis of leprosy is currently based on the appearance of clinical signs and requires expert clinical, as well as labor-intensive and time-consuming laboratory or histological, evaluation. For the purpose of developing an effective, simple, rapid, and low-cost diagnostic alternative, we have analyzed the serologic antibody response to identify Mycobacterium leprae proteins that are recognized by leprosy patients. More than 100 recombinant antigens were analyzed in a protein array format to select those with discriminatory properties for leprosy diagnosis. As expected, multibacillary leprosy patients recognized more antigens with stronger antibody responses than paucibacillary leprosy patients. Our data indicate, however, that multibacillary patients can be distinguished from paucibacillary patients, and both of these groups can be segregated from endemic control groups. We went on to confirm the diagnostic properties of antigens ML0405 and ML2331 and the LID-1 fusion construct of these two proteins by enzyme-linked immunosorbent assay. We then demonstrated the performance of these antigens in rapid test formats with a goal of developing a point-of-care diagnostic test. A serological diagnostic test capable of identifying and allowing treatment of leprosy could reduce transmission, prevent functional disabilities and stigmatizing deformities, and facilitate leprosy eradication.     

Review of egg-related salmonellosis and reduction strategies in United States,Australia, United Kingdom and New Zealand

Globally, Salmonella enterica subsp. enterica is one of the most commonly reported causes of foodborne illness in humans. Contaminated food products of animal origin, particularly egg and egg products are frequently implicated in outbreaks of human salmonellosis. Salmonella enteritidis is frequently involved in egg and egg products-associated foodborne outbreaks in the USA and UK. However, in Australia and New Zealand, human infections caused by this serovar occur as a result of infection acquired while overseas travel, with Salmonella typhimurium being a predominant cause of local foodborne outbreaks. In this paper, an overview of Salmonella epidemiology on laying farms, egg-related Salmonella outbreaks in humans, and regulatory practises to control Salmonella across USA, UK, Australia and New Zealand is provided. Considering the estimated production of eggs in the USA, UK, Australia and New Zealand in 2015, the risk of foodborne illness in general is quite low for humans consuming eggs. Salmonella diagnostics, reporting and surveillance systems have improved over the years and will continue to improve in the years to come. However, given the number of different emerging Salmonella serovars a regular review of Salmonella control strategies from farm to fork is required.     

Endotoxemia increases the clearance of mPEGylated 5000-MW quantum dots as revealed by multiphoton microvascular imaging

Imaging the microcirculation is becoming increasingly important in assessing life-threatening disease states. To address this issue in a highly light absorbing and light scattering tissue, we use laser scanning multiphoton microscopy and fluorescent 655-nm 5000-MW methoxy-PEGylated quantum dots to image the functional microcirculation deep in mouse hind limb skeletal muscle. Using this approach, we are able to minimize in vivo background tissue autofluorescence and visualize complete 3-D microvascular units, including feeding arterioles, capillary networks, and collecting venules to depths of 150 to 200 microm. In CD1 mice treated with lipopolysaccharide to model an endotoxemic response to bacterial infection, we find that these quantum dots accumulate at microvascular bifurcations and extravasate from the microcirculation in addition to accumulating in organs (liver, spleen, lung, and kidney). The quantum dots are cleared from the circulation with a first-order elimination rate constant seven times greater than under normal conditions, 1.6+/-0.06 compared to 0.23+/-0.05 h(-1), P<0.05, thereby reducing the imaging time window. In vitro experiments using TNFalpha treated isolated leukocytes suggest that circulating monocytes (phagocytes) increased their nonspecific uptake of quantum dots when activated. In combination with multiphoton microscopy, quantum dots provide excellent in vivo imaging contrast of deep microvascular structures.     

Preparation of a Noncytotoxic Hemocompatible Ion Gel by Self‐Polymerization of HEMA in a Green Deep Eutectic Solvent

Self‐polymerization of optimized amount of 2‐hydroxyethylmethacrylate (HEMA) in a natural deep eutectic solvent (NADES) is obtained by the complexation between choline chloride and fructose (ChCl:Fruc. 2:1) in the presence of indomethacin, an anti‐inflammatory drug resulted formation of a drug‐immobilized ion gel. The ion gel thus obtained is found to be stable in gastrointestinal fluid pH (GIF) and sustained release in simulated lower intestinal fluid for 20 h (88% release) at pH 6.8 and 96% release at pH 7.4 for 15 h (colon & blood pH) at physiological human body temperature (37 °C) of the drug is recorded. The drug can be stored in the gel matrices at room temperature for long durations such as 6 months without any degradation. The ion gel of poly‐HEMA obtained in the NADES does not appear to inhibit the growth of mammalian cells in vitro and is found to be compatible to human blood thus implying toward their potential for therapeutic applications as pH‐sensitive hydrophobic drug carrier for oral application as well as in tissue engineering.