immunoglobulin and histamine-sensitivity response of mice to live Bordetella pertussis

Mice injected intranasally (it.n.) and intraperitoneally (i.p.) with a nonlethal dose (2.5 x 10(5) colony-forming units) of live Bordetella pertussis were examined for 50 days for infection, respiratory tract immunoglobulins (Ig), changes in serum Ig, and histamine sensitivity. With mice infected it.n., respiratory infection markedly declined between day 20 and day 30. Ig classes (A, G(1), G(2a), G(2b), but no M), which had specificity for B. pertussis, were present in tracheobronchial wash (TBW) by day 15; by day 50, TBW immunodiffusion and immunoelectrophoretic precipitin bands were more intense. A sharp rise in serum IgA after day 30 was the only significant change relative to controls among the five serum Ig examined. A high degree of histamine sensitivity developed by day 15 to 20 and persisted for the 50 days. With mice inoculated i.p., no bacteria were recovered, no Ig or only traces were found in TBW and IgA only was specific, and no significant changes in the serum Ig relative to controls occurred. Histamine sensitivity developed somewhat more slowly and to a lesser degree than in it.n.-injected mice but persisted for the 50 days. A similar small number of killed bacteria (pertussis vaccine) injected it.n. or i.p. likewise induced slowly developing histamine sensitivity in contrast to published reports of 4 to 5 day peak sensitivity and decline following i.p. injection of 10(9) or more killed bacteria.     

Real Time-PCR HBV-DNA Analysis: Significance and First Experience in Armed Forces

Background

HBV DNA quantitation is used extensively world wide for the diagnosis and monitoring of treatment of Hepatitis B virus (HBV) infection. However, it has still to be popular in India. The aim of this study was to quantitate HBV – DNA by Real time – PCR method in Hepatitis B and in immuno-compromised patients, to compare the results with HBeAg detection and to monitor the response to therapy of chronic Hepatitis B patients to antivirals.

Methods

Ninety one serum samples of Hepatitis group of patients (all HBsAg positive), 41 samples from immuno-compromised patients (all HBsAg negative) and 49 patients of Chronic Hepatitis B group (all HBsAg positive) were the subjects of this first ever study in Armed Forces. Twenty serum samples from healthy volunteers and non-hepatitis B patients served as negative controls. The amplification detection was carried out in a Rotor-Gene 2000-sequence detector

Results

Amongst Hepatitis B group, 33% (30/91) of the samples were positive for HBV-DNA and 26% (24/91) of samples were positive for HBeAg. In the immuno-compromised group of patients 14.6% (6/11) of samples were positive for HIV-DNA and 9.7% (4/41) were positive for HBeAg. Of the Chronic Hepatitis B patients on treatment, all (100%) were positive by HBV-DNA, whereas 29/49 (59.2%) were positive by HBeAg before treatment. After treatment with antivirals, 06/49 (12.2%) were positive by both tests and 11/49 (22.5%) were positive only by HBV-DNA. 32/49 (65.3%) patients became negative serologically after therapy.

Conclusion

HBeAg status did not necessarily reflect HBV-DNA level in the serum, as 10/91 (11%) in the Hepatitis B group, 2/41 (4.9%) in the immuno compromised group and 20/49 (40.8%) patients in the Chronic Hepatitis B group were positive for HBV-DNA but negative for HBeAg. HBV-DNA was not found to be positive amongst any of the negative controls. Real time – PCR is a sensitive and reproducible assay for HBV-DNA quantitation and may be started in Armed Forces referral centers in the near future.Key Words: Real time – PCR, Chronic Hepatitis B, HBV – DNA, Antivirals     

Assessing relationship betrayals

Relationship betrayals, such as sexual and emotional infidelity, are commonly encountered in the practice of psychotherapy, yet clinicians often report difficulties in assessing and treating them. In this article we offer suggestions for assessing relationship betrayals. We address the definition of relationship betrayals, methods of assessing relationship betrayals, when and whom to assess, confidentiality, and assessing related clinical concerns. We illustrate with case vignettes the assessment and treatment of betrayals in close relationships.     

A new video image analysis system to study red blood cell dynamics and oxygenation in capillary networks

OBJECTIVE: The authors present a Measurement and Analysis System for Capillary Oxygen Transport (MASCOT) to study red blood cell (RBC) dynamics and oxygenation in capillary networks. The system enables analysis of capillaries to study geometry and morphology and provides values for capillary parameters such as diameter and segment length. It also serves as an analysis tool for capillary RBC flow characteristics, including RBC velocity, lineal density, and supply rate. Furthermore, the system provides a means of determining the oxygen saturation of hemoglobin contained within RBCs, by analysis of synchronized videotapes containing images at two wavelengths, enabling the quantification of the oxygen content of individual RBCs. METHODS: Video recordings of RBC flow at two wavelengths, 420 nm (isosbestic) and 436 nm (oxygen sensitive), are made using a dual camera video microscopy system. The 420-nm recording is used to generate images based on the variance of light intensity fluctuations that help to identify capillaries in a given field of view that are in sharp focus and exhibit flow of individual RBCs separated by plasma gaps. A region of interest enclosing the desired capillary is defined and a fixed number of successive video frames at the two wavelengths are captured. Next a difference image is created, which delineates the RBC column, whose width is used to estimate the internal diameter of the capillary. The 420-nm images are also used to identify the location and centroid of each RBC within the capillary. A space-time image is generated to compute the average RBC velocity. Lineal density is calculated as the number of RBCs per unit length of a capillary segment. The mean optical density (OD) of each RBC is calculated at both wavelengths, and the average SO(2) for each cell is determined from OD(436)/OD(420). RESULTS AND CONCLUSIONS: MASCOT is a robust and flexible system that requires simple hardware, including a SGI workstation fitted with an audio-visual module, a VCR, and an oscilloscope. Since the new system provides information on an individual cell basis from entire capillary segments, the authors believe that results obtained using MASCOT will be more accurate than those obtained from previous systems. Due to its flexibility and ease of extension to other applications, MASCOT has the potential to be applied widely as an analysis tool for capillary oxygen transport measurements.     

Embryonic behavior,hatching and neuromuscular development in the chick following a transient reduction of spontaneous motility and sensory input by neuromuscular blocking agents

Embryos were immobilized with neuromuscular blocking agents for one to four days between days 10 and 15 of incubation. This treatment reduces spontaneous motility, as well as movement-initiated proprioceptive and cutaneous stimulation. Although the major aim of these experiments was to determine the effects of such treatment on subsequent behavioral development, several indices of neuromuscular and general morphological development were also examined. A single injection of curare on day 10 continues to depress spontaneous motility for virtually the entire remaining incubation period. This effect is due to the persistence of unmetabolized curare in the closed system of the egg. When a comparable dose of a rapidly metabolized neuromuscular blocking agent (succinylcholine) is given, this long term behavioral depression is not found. Embryos treated with SC can remain totally immobilized for up to 60 hours with no apparent repercussions on subsequent behavior; spontaneous embryonic motility, reflex sensitivity, hatching, and several general posthatching behaviors, all appeared normal following such treatment. Embryos immobilized for as little as 48 hours developed joint malformations and were retarded in general growth by about one day whereas 24 hours of paralysis was not sufficient to induce these effects. Twenty-four hours of total paralysis from days 10 to 11, plus a 40–50% reduction in motility until days 15 or 16, does not appear to cause any abnormalities in muscle or spinal cord development. Muscle histology, motor endplates, cell number in the spinal cord and choline acetyltransferase and acetylcholinesterase activity in spinal cord and muscle were all comparable to controls. An 80–90% reduction in motility on days 11 through 15, however, induces an apparent alteration of the intensity and distribution of histochemically demonstrable AChE in the anterior and posterior latissimus dorsi muscles. The present findings suggest that the suppression of overt motility for a period comprising ca. 5–12% of the total incubation time (21 days) does not modify subsequent behavioral development or the underlying neurogenetic mechanisms. Though these findings provide some support for the notion that the embryonic nervous system develops in forward reference to and without benefit from function or sensory input, only a more complete reduction of neural function, especially CNS activity, can provide a critical test of this concept.