Reed-Sternberg cells in Hodgkin's cell lines HDLM, L-428, and KM-H2 are not actively replicating: lack of bromodeoxyuridine uptake by multinuclear cells in culture

We compared the proliferation of mononuclear and multinuclear cells in four Hodgkin's cell lines, HDLM-1, HDLM-1d, L-428, and KM-H2, by examining their capacity to incorporate bromodeoxyuridine (BrdUrd) into nuclei. Approximately 5% of all cells in HDLM-1 cultures had two or more nuclei, a characteristic of Reed-Sternberg (RS) cells. Unlike mononuclear Hodgkin's (H) cells, these RS cells exhibited no uptake, or only minimal uptake of BrdUrd, suggesting that they did not replicate actively. Cytogenetic study showed that 25% of the HDLM-1 cells contained a tetraploid (4X) set of chromosomes with a characteristic two-peak distribution. Following treatment of HDLM-1 cells with phorbol ester, the percentages of 4X cells and RS cells increased to 50% and 12%, respectively. This increase in RS cells was not likely to be due to cell fusion as shown by the absence of hybridization of BrdUrd- positive and -negative nuclei. Phorbol ester has a short-term effect of blocking the exit of cells from G1 into S phase, but no effect on the transition from S phase to G2/M phase. The block is more prominent in 2X cells than in 4X cells, which may explain the increase in percentage of 4X cells in phorbol ester-treated cultures. In addition, phorbol ester induced the differentiation of H-RS cells, which was accompanied by loss of the marker HeFi-1 from the cell surface. Approximately one third of the RS cells did not express HeFi-1, or expressed only minimal amounts. The findings led us to the following conclusions: (1) The 4X cells probably are formed from 2X H cells as a result of disturbed cytokinesis, but not a cell fusion. (2) A considerable number of 4X cells were H cells, because the number of 4X cells consistently exceeded that of RS cells. (3) Since mitotic figures are extremely rare in RS cells and these cells did not show active BrdUrd uptake, the increased number of RS cells must also be a consequence of disturbed cytokinesis of H cells or a result of nuclear transformation (twisting, convolution, or separation of the nucleus) in H cells. (4) Most RS cells lose their proliferating capacity and some RS cells may undergo further differentiation. Uptake of BrdUrd and phorbol ester induction were also studied on the other three H-RS cell lines, HDLM-1d, L-428, and KM-H2, with results similar to those for HDLM-1.     

Bacterial renal infection: role of CT

The imaging studies done on 62 patients hospitalized for acute renal infections were retrospectively reviewed. Thirty-six (58%) had one or more abscesses, 17 (27%) had focal or diffuse acute bacterial nephritis, five (8%) had pyonephrosis, and four (6%) had pyelonephritis. All had prolonged fever (greater than or equal to 72 hours) and leukocytosis. Among 25 patients examined with both ultrasound (US) and computed tomography (CT), US failed to depict three of five (60%) cases of acute bacterial nephritis and seven of 15 (47%) intrarenal and extrarenal abscesses. One renal abscess was misdiagnosed as a tumor at CT. US is not an adequate screening test for detecting lesions that may require invasive therapy. CT is more sensitive for the detection of acute renal inflammatory disease and for defining the extent of disease for planning of radiologic or surgical intervention.     

Treatment of Casualties in a Forward Hospital of Indian Army : Nine year Experience

Background

To analyze the outcome of the management of casualties in a level II trauma centre of a forward hospital of Armed Forces over a nine year period. Retrospective analysis of all casualties received in a single forward hospital of Indian Army was carried out.

Method

During 9 years (1990-1998), a total of 5737 casualties were received in a single level II zonal hospital of the Army in a forward area. Majority of the injuries were caused by bullets, or by fragments of improvised explosive devices. A policy of aggressive resuscitation and early primary repair of injuries was followed. General surgeons routinely performed craniotomies, thoracotomies, laparotomies, stabilization of fractures by fixators and repair of vascular injuries.

Result

38% of patients had injuries to several body parts (polytrauma), resulting in a total of 8578 injuries. Region-wise distribution of injuries was as follows : 14.2% head and neck injuries, 13.3% chest wounds, 13.5% abdominal injury and 59% extremity wounds. The overall mortality rate was 3.6%. The complication rate was about 7% with infection as the major complication. The results of primary repair of colonic injuries were similar to those of staged repairs. The results after primary closure of war wounds were better than those treated with delayed primary closure in selected cases.

Conclusion

Prompt evacuation, speedy resuscitation and early definitive repair of war injuries results in low mortality and morbidity. A motivated and dedicated team and adequate availability of blood and ancillary services adds to the excellent outcome. The policy of primary repair of colonic and selected soft tissue injuries appears justified in selected cases.Key Words: Casualties, Improvised Explosive Devices, Militancy, Trauma     

Norms of the Rorschach Test for Indian Subjects

Background

The clinical utility of the Rorschach test in Indians is hampered by the absence of reliable normative data. Method : The Rorschach by Dlopfer''s method was administrated to 1256 subjects consisting of 300 normal army personnel, 300 normal civilians, 250 schizophrenics, 300 neurotics and 106 patients with organic disorders.

Results

The Rorschach protocols of normal Indian army personnel and normal civilians showed significant differences from one another and also from the western norms. These differences are culturally determined and are not indicative of low intelligence or psychopathology. Patients with schizophrenia, neurosis, head injury and epilepsy show significant differences from the records of normal subjects. The protocols of army schizophrenics show significant deviations from those of normal army personnel and these changes revert to normal with clinical recovery.

Conclusion

The Rorschach test is not a culture fee test as claimed earlier. In view of the differences from Western norms, Rorschach protocols of Indians should be interpreted using the norms for Indians. In the case of army personnel the norms for army personnel should be used. While the use of the Rorschach to study the personality patterns of normal individuals and as an aid to clinical diagnosis was strongly supported, the findings of the study indicate that the test can also be employed to assess therapeutic response of patients with schizophrenia.Key Words: Rorschach test, norms, schizophrenia, neurosis     

Specific and sensitive detection of Neisseria gonorrhoeae in clinical specimens by real-time PCR

Early diagnosis of Neisseria gonorrhoeae infections is important with regard to patients' health and infectivity. We report the development of a specific and sensitive TaqMan assay for the detection of N. gonorrhoeae in clinical samples. The target sequence is a 76-bp fragment of the 5' untranslated region of the opa genes that encode opacity proteins. A panel of 448 well-defined N. gonorrhoeae isolates was used to evaluate and optimize the assay. The method employs two minor-groove binding probes, one of them recognizing a newly identified sequence in the opa genes. Testing a large panel of related and unrelated microorganisms revealed that other Neisseria strains and other microorganisms tested negative in the opa test. With a lower detection limit of one genome per reaction, the opa test appeared more sensitive than both the COBAS AMPLICOR (Roche Diagnostics Nederland BV, Almere, The Netherlands) and a LightCycler 16S rRNA test. Analysis of a panel of 122 COBAS AMPLICOR-positive samples revealed that 68% were negative in both the 16S rRNA test and the opa assay (confirming that the COBAS AMPLICOR test produces false positives), while 30% were positive in both assays. Three samples were opa positive and 16S rRNA negative, which may be due to the higher sensitivity of the opa assay. We conclude that the opa gene-based real-time amplification assay offers a sensitive, specific, semiquantitative, and reliable assay suitable for the detection of N. gonorrhoeae in clinical specimens and/or for confirmation of less specific tests.     

A randomised, placebo controlled trial of inhaled salbutamol and beclomethasone for recurrent cough

AIMS: To test the hypothesis that inhaled salbutamol or beclomethasone will reduce the frequency of cough in children with recurrent cough. A secondary aim was to determine if the presence of airway hyperresponsiveness (AHR) can predict the response. DESIGN: Randomised, double blind, placebo controlled trial. METHODS: During a coughing phase, 43 children (age 6-17 years) with recurrent cough were randomised to receive inhaled salbutamol or placebo (phase I) for 5-7 days and then beclomethasone or placebo (phase II) for 4-5 weeks, and in a subgroup of children for 8-9 weeks. The children used an ambulatory cough meter, kept cough diaries, and performed the capsaicin cough sensitivity, hypertonic saline bronchoprovocation, and skin prick tests. RESULTS: Salbutamol or beclomethasone had no effect on cough frequency or score, irrespective of the presence of AHR. CONCLUSIONS: Most children with recurrent cough without other evidence of airway obstruction, do not have asthma and neither inhaled salbutamol nor beclomethasone is beneficial.     

Gastric tonometry in septic shock

OBJECTIVES: To investigate the prognostic value of intramucosal pH (pHi) and the relation among pHi, arterial pH, base excess, and lactate in children with septic shock. DESIGN: Children admitted to the paediatric intensive care unit with a diagnosis of septic shock were prospectively enrolled. A gastrointestinal tonometer (Tonometrics Division, Instrumentarium Corporation, Helsinki, Finland) was placed into the stomach and intramucosal pH, arterial pH, base deficit, and lactate were measured on admission and six hours later. Sequential data were analysed on 24 patients (17 survivors, seven non-survivors), median age 46 months (range: 2.8-168 months). RESULTS: Median pHi on admission was 7.39 (interquartile range 7.36-7.51) in survivors compared with 7.2 (interquartile range 7.18-7.35) in non-survivors (p = 0.01). There was no significant difference in arterial pH, base excess, or lactate among survivors and non-survivors. Admission pHi < 7.32 predicted mortality with sensitivity (57%), specificity (94%), and positive predictive value (80%). Patients with admission pHi < 7.32 who failed to improve > or = 7.32 within six hours (n = 3) had 100% mortality. CONCLUSION: In children with septic shock the admission pHi is significantly lower in non-survivors. pHi is a better prognostic indicator of mortality than either standard acid-base values or lactate. pHi < 7.32 that does not improve within six hours is associated with a poor prognosis.     

Impact of zinc supplementation on persistent diarrhoea in malnourished Bangladeshi children

To evaluate the impact of zinc supplementation on the clinical recovery and body weight of children with persistent diarrhoea, a randomized, double-blind, controlled trial was conducted in 190 children with persistent diarrhoea aged between 3 and 24 months. Children were randomly allocated to receive either zinc (20 mg d⁻¹) syrup with multivitamin (2 × RDA) or multivitamin alone in three divided daily doses for 2 weeks. The trial was conducted in a diarrhoeal disease hospital in Dhaka, Bangladesh. Duration until clinical recovery (d), impact on body weight and serum zinc level after 2 weeks of zinc supplementation were recorded. The duration of illness was significantly reduced (33%) with zinc supplementation among children who were underweight (≤70% wt/age, p = 0:03). Supplemented male children also had a significant reduction (27%) in duration for recovery compared with unsupplemented children ( p = 0:05). From baseline to convalescence, zinc-supplemented children maintained their serum zinc concentration (13.4 vs 13.6/ μ mol l⁻¹), whereas unsupplemented children had a decrease in serum zinc after the 2 weeks of diarrhoea (13.6 vs 11.8 μ mol l⁻¹, p < 0:03). The mean body weight of the children in the supplemented group was maintained (5.72 vs 5.70 kg, p = 0:62) during hospitalization, unlike that of the control group, in which there was a reduction in body weight (5.75 vs 5.67 kg, p = 0:05). Five children in the unsupplemented group and one child in the zinc-supplemented group died during the 2 weeks of supplementation ( p = 0:06). Zinc supplementation in persistent diarrhoea significantly reduced the length of the recovery period in malnourished children and prevented a fall in body weight and serum zinc concentration, indicating that zinc is a beneficial therapeutic strategy in this high-risk childhood illness.