Field evaluation of ELISA using Wuchereria bancrofti mf ES antigen for bancroftian filariasis

An enzyme-linked immunosorbent assay using Wuchereria bancrofti microfilarial excretory-secretory antigen was used in field studies to screen blood samples collected on filter-paper from persons residing in areas endemic for bancroftian filariasis. This assay system, when compared with examination of night wet blood smears for microfilariae, gave a relative sensitivity of 98% and a relative specificity of 86%. Daytime blood samples can also be used in this test, which can thus replace tedious examination of night blood samples in field surveys in endemic areas.     

Fractionation and characterization of urinary filarial antigen in bancroftian filariasis

Urine samples from microfilaraemic patients were concentrated and fractionated by gel chromatography on Ultrogel AcA 44. Four protein fractions labelled as UFA C1, UFA C2, UFA C3 and UFA C4 were tested for filarial antigenicity by sandwich ELISA. UFA C1 and UFA C2 showed antigenic activity. On further analysis by SDS-PAGE, UFA C1 and UFA C2 showed antigenic components with MW ranging from 10.4 K to 123 K. UFA C1-1 and UFA C2-2 showed high antigen titre in ELISA. Urinary albumin was observed as a major component in UFA C2. Absorption of albumin from UFA C2 enhanced its antigenic activity considerably. As little as 0.01 pg antigenic protein per test was found to be sufficient for the detection of filarial antibody in ELISA. Biochemical characterization indicated a glycoprotein nature of UFA C2.     

Automated system for detailed measurement of respiratory mechanics

Objective. The mechanical properties of the respiratory system (i.e., elastance and resistance) depend on the frequency, tidal volume, and shape of the flow waveform used for forcing. We developed a system to facilitate accurate measurements of elastance and resistance in laboratory and clinical settings at the frequencies and tidal volumes in the physiologic range of breathing.Methods. A personal computer (PC) is used to drive a common clinically used ventilator while simultaneously collecting measurements of airway flow, airway pressure, and esophageal pressure from the experimental subject or animal at different frequencies and tidal volumes. Analysis analogous to discrete Fourier transform at the fundamental frequency (i.e., ventilator setting) is used to calculate elastances and resistances of the total respiratory system and its components, the lungs and the chest wall. We have shown that this analysis is independent of the high-frequency harmonics that are present in the waveform from clinical ventilators.Results. The system has been used successfully to make measurements in anesthetized/paralyzed dogs and awake or anesthetized human volunteers in the laboratory, and in anesthetized humans in the operating room and intensive care unit. Elastances and resistances obtained with this approach are the same as those obtained during more controlled conditions, e.g., sinusoidal forcing. Conclusions. Accurate, standardized measurements of lung and chest wall properties can be obtained in many settings with relative ease with the system described. These properties, and their frequency and tidal volume dependences in the physiologic range, provide important information to aid in the understanding of changes in respiratory function caused by day-to-day conditions, clinical intervention and pathologies.The authors thank Colin Mackenzie for his suggestions throughout the experiments.This work was supported by the National Heart, Lung, and Blood Institute grants HL-33009 and HL-44128.     

Comparative Evaluation of Clinical Efficacy of Leukocyte-Rich Platelet-Rich Fibrin with Advanced Platelet-Rich Fibrin in Management of Gingival Recession Defects: A Randomized Controlled Trial

BackgroundThe aim of this research was to determine and compare the clinical efficacy of leukocyte platelet-rich fibrin (L-PRF) and advanced platelet-rich fibrin (A-PRF) in combination with coronally advanced flap (CAF) in the treatment of gingival recession defects.MethodsSystemically healthy subjects presenting with 30 Miller''s class I or II gingival recession defects in maxillary anteriors and premolars, were treated with either CAF + L-PRF or CAF + A-PRF. Clinical parameters such as recession height (RH), width, probing pocket depth, clinical attachment level (CAL), keratinized tissue height (KTH), and width of attached gingiva (WAG) were measured at baseline, 3, and 6 months. Gingival biotype was evaluated at baseline and 6 months post-surgery. Mean root coverage percentage (MRC%) was evaluated at 3 and 6 months.ResultsStatistically significant reduction in mean RH was observed from baseline (2.53 ± 0.74 mm, 2.63 ± 0.82 mm) to 6 months (0.87 ± 0.83 mm, 0.53 ± 0.91 mm) in CAF + L-PRF and CAF + A-PRF groups, respectively. The MRC% achieved at 6 months was 67.20 ± 32.81 in the CAF + L-PRF group and 81.66 ± 28.21 in the CAF + A-PRF group. Statistically significant gain in CAL, WAG, and KTH was observed in both therapeutic groups (p < 0.05). Intergroup analysis revealed no statistically significant differences among study parameters between groups at any time point (p > 0.05).ConclusionBased on the findings of this study, both L-PRF and A-PRF may be suggested as viable treatment options for the management of gingival recession in maxilla.     

Design,Synthesis, Antioxidant,and Anti‐Breast Cancer Activities of Novel Diethyl(alkyl/aryl/heteroarylamino)(4‐(pyridin‐2‐yl)phenyl)methylphosphonates

A series of new diethyl(alkyl/aryl/heteroarylamino)(4‐(pyridine‐2‐yl)phenyl)methylphosphonates ( 4a–t ) were synthesized via three‐component Kabachnik–Field's reaction of 4‐(pyridin‐2‐yl)benzaldehyde, diethylphosphite and various primary amines, catalyzed by cupric acetate monohydrate [Cu(OAc)₂ · H₂O] under solvent‐free and microwave irradiation conditions. Their computational docking analysis supported them as good therapeutic agents to the breast cancer aromatase enzyme and ascertained 4a , 4h , 4m , 4n , and 4t as potential molecules with good binding affinities varying from ?9.0 to ?9.6 kcal/mol and containing the 4‐(pyridine‐2‐yl)phenyl moiety as a pharmacophore. Their in vitro screening performed for the anti‐cell proliferation activity against MBC‐MCF7 cells by MTT and Trypan blue assays confirmed 4m , 4n , and 4q as promising compounds to sustain a low percentage of cell viability at 20 µg/mL concentration. These compounds were also evaluated for their antioxidant activity by the DPPH method and the results established that compounds 4m , 4n , and 4q show around 10% higher activity than the standard antioxidant ascorbic acid.     

Improved laboratory diagnosis of tuberculosis – The Indian experience

Tuberculosis (TB) is the leading cause of death worldwide attributable to a single infectious disease agent. India has more new TB cases annually than any other country. In 2008, India accounted for a fifth of the estimated 9.4 million TB cases globally. There is an overwhelming need for improving TB diagnostics in India through the use of cost effective, patient-friendly methods appropriate to different tiers of the country health system. Substantial progress has been made in India in the field of TB diagnosis and serious efforts have been made to herald the development of diagnostic tests for pulmonary TB, extra pulmonary TB and MDR-TB. Diverse approaches have been attempted towards improving smear microscopy, rapid culture and for differentiation between the Mycobacterium tuberculosis complex and non-tuberculous mycobacteria. Several laboratories have developed in-house PCR assays for diagnosing TB with high accuracy. Approaches for distinguishing M. tuberculosis and/or Mycobacterium bovis infection and disseminated Mycobacterium avium complex infection in HIV-AIDS patients have also been described. Serological tests to detect antigens or antibodies to M. tuberculosis specific components by using cocktails of Excretory/Secretory protein antigens, Ag85 complex antigens, Hsp 65 antigen, RD1 antigens and Rapid Reverse Line Blot Hybridization assays to detect MDR-TB (mutations to rifampicin, isoniazid and streptomycin) have also been developed. Other methods like measurement of adenosine deaminase activity and use of luciferase reporter phages have also been explored for TB diagnosis. These advances in the Indian context are detailed in the present chapter. The validation and application of these methods in laboratory and public health settings is likely to result in improved TB diagnosis and contribute to effective disease management in India.