Crystallization of agGST1-6, a recombinant glutathione S-transferase from a DDT-resistant strain of Anopheles gambiae.

Glutathione S-transferases (GSTs) belong to a family of detoxification enzymes that conjugate glutathione to various xenobiotics, thus facilitating their expulsion from the cell. GST activity is elevated in many insecticide-resistant insects, including the DDT-resistant malaria vector Anopheles gambiae. Crystals of the recombinant form of a GST from A. gambiae, agGST1-6, have been grown in at least five different crystal forms, with a broad range of diffraction resolution limits. A complete 2.0 A data set has been collected on a C-centered orthorhombic crystal form with unit-cell parameters a = 99.0, b = 199.4, c = 89.6 A. A search for heavy-atom derivatives has been initiated, along with phase-determination efforts by molecular replacement.

Lactate dehydrogenase release assay from Vero cells to distinguish verotoxin producing Escherichia coli from non-verotoxin producing strains.

The Vero cell assay presently used for virulence testing of verotoxigenic Escherichia coli (VTEC) requires at least 48-96 h where cytotoxicity effects are examined under a microscope. Here, a complimentary rapid assay was developed that measures endogenous lactate dehydrogenase (LDH) release from Vero or HEp-2 cells as an indicator of cytotoxicity. Toxin preparations from 24 VTEC strains induced 36-89% LDH from Vero cells and 15-62% LDH from HEp-2 cells in 12-16 h. A verotoxin-positive but enterohemolysin negative strain also showed a similar cytotoxicity effect. In contrast, three VT-negative strains caused only 13-16% LDH from Vero cells and 1-7% LDH from HEp-2 cells. Five presumptive E. coli isolates from naturally contaminated food and clinical sources did not induce significant LDH release from either cell lines. PCR analysis confirmed the presence of vt1 or vt2 genes in E. coli showing positive LDH values. Similarly, RiboPrinter analysis confirmed and identified the test strains as E. coli except for two meat isolates, which were identified as Hafnia alvei. Cytopathic effects of toxin preparations from VTEC revealed severe lysis, vacuole formation and death in Vero cells and multiple vacuoles and cell elongation in HEp-2 cells. The colorimetric cytotoxicity assay described here can provide quantitative data for determining the virulence potential of verotoxigenic E. coli in 12-16 h.

Preparing students with special needs for reintegration: curriculum-based measurement's impact on transenvironmental programming.

The original intent of special education pull-out services was to remove students from general education temporarily for intensive, individualized instruction in deficit areas. In many school districts, however, it seems that once students participate in pull-out services, there is little effort to return them to general education. One explanation for this is that not much is known about how best to prepare students for movement up the cascade of services, including reintegration into the mainstream. A promising reintegration approach is transenvironmental programming (TP). Research suggests that supplementing TP with continuous progress monitoring, specifically via curriculum-based measurement (CBM), better prepares students for reintegration than either TP or CBM alone. Findings from the present study provide a possible explanation: Results suggest that special educators using TP who receive CBM information about their students' academic progress are more likely to plan and implement academic interventions in preparation for students' transition than are those special educators who do not receive this information. This supports the hypothesis that the relatively greater success of TP + CBM is due to increased attention to academic preparation. Implications for practice and future research are discussed.

Effect of leuprolide acetate in patients with moderate to severe functional bowel disease. Double-blind, placebo-controlled study.

Moderate to severe functional bowel disease results in debilitating abdominal pain, nausea, intermittent vomiting, early satiety, bloating, abdominal distension, and/or altered bowel habits. Because it occurs approximately 20-30 times more frequently in women than in men and its symptoms often coincide with the menstrual cycle, we hypothesized that reproductive steroids may antagonize diseased nerves of the gastrointestinal tract, enhancing the expression of symptoms. No effective or consistent therapy has existed for these patients. We prospectively investigated the effect of a gonadotropin-releasing hormone analog, leuprolide acetate, in 30 women with symptoms of moderate to severe functional bowel disease. The study was phase II, randomized, double blind, and placebo controlled. Lupron Depot 3.75 mg (which delivers a continuous low dose of drug for one month) or placebo were given intramuscularly monthly for three months. Symptom scores were assessed at each four-week visit. Follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone levels were assessed before and after therapy. Patients treated with low-dose leuprolide improved progressively and significantly in scores for nausea, vomiting, bloating, abdominal pain, and early satiety, and for overall symptoms (P < 0.01-0.05). All hormone levels decreased significantly (P < 0.05) except luteinizing hormone (P = 0.054).

Effect of leuprolide acetate in patients with functional bowel disease. Long-term follow-up after double-blind, placebo-controlled study.

We initially investigated the effects of a gonadotropin-releasing hormone analog, leuprolide acetate, in 28 patients with moderate to severe functional bowel disease in a phase-II, randomized, double-blind, and placebo-controlled study using Lupron Depot 3.75 mg (which delivers a continuous low dose of drug for one month) or placebo given intramuscularly. After completing that 12-week study period during which their symptoms had improved significantly (P < 0.01-0.5), the 28 patients were allowed to continue receiving leuprolide acetate; they were monitored for an additional 40 weeks. Of those 28, 25 (89%) finished the 52-week treatment. Drug administration was changed from the monthly low-dose form of leuprolide acetate to a daily subcutaneous dose that was gradually increased from 0.5 mg daily to an effective therapeutic dose (1.0-1.5 mg). All subjects received estrogen replacement during this period. Continued use of leuprolide acetate at maximum therapeutic dosage and over longer periods of time produced even more striking and significant changes in the disabling and debilitating symptoms of functional bowel disease. Nausea, abdominal pain, early satiety, anorexia, and abdominal distension decreased markedly (P < 0.0001) and vomiting was also reduced (P < 0.01) more than in the short-term, low-dosage, double-blind study. Combined total symptom scores and overall assessment also changed significantly in the long-term phase (both P < 0.0001).

Simply a case of good practice. Avoiding catheter-related sepsis in total parenteral nutrition.

Catheter-related sepsis is a well-known complication associated with the administration of total parenteral nutrition. A strict aseptic approach, supplemented with a knowledge of how the infection occurs, will do much to avoid this problem.

Monocondylar fractures of the femur: a review of 13 patients.

We reviewed 13 patients with monocondylar fractures of the femur. Undisplaced fractures may require extensive radiological investigation to visualize the fracture. Anatomically reduced fractures may displace when treated non-operatively. Joint line incongruity and resultant callus formation produces an abnormal condylar profile leading to early secondary osteoarthritis. We recommend internal fixation in all patients with monocondylar fractures.

Internal sterilization using the Falope ring.

PIP: The article reports on results obtained using a Falope ring for interval sterilization in 2 series of patients, one of 263 patients, and another of 1583 patients, both studies conducted between 1976-78. There were only 2 pregnancies in the 1st series; over 90% of patients were satisfied with the procedure. There were 3 pregnancies in the 2nd series, and 2 cases of bleeding requiring laparotomy. Complications were minimal in both series. Interval sterilization with a Falope ring can be recommended over electrocoagulation when there are no adherent tubes, and it has the added advantage of being, at least potentially, more reversible than other procedures.^ieng

Chondrosarcoma of the bones of the hand.

Nineteen chondrosarcomas are reported arising in proximal phalanges or metacarpal bones of the hand mainly in elderly patients, predominantly women. The usual clinical presentation was of a progressively painful large tumour, often arising in a dormant lesion near the metacarpo-phalangeal joint. Radiologically most showed some bone expansion with a poorly defined area of destruction and a considerable soft-tissue swelling. Histologically, malignancy was usually obvious, but confusion might arise from the inclusion of bland areas of chondromatous tissue that probably represented the original lesion. Four tumours, initially curetted and grafted, recurred locally and necessitated amputation of the digit or ray. Amputation was the primary treatment for fourteen other tumours and was curative except in one patient who eventually needed amputation through the forearm for a large second recurrence. One tumour was satisfactorily controlled by excision of the affected phalanx. None of these nineteen tumours is known to have metastasised. Correct treatment implies a carefully considered balance between conservation of function and complete removal of all tumour tissue.