Carbohydrate supplementation and the lymphocyte proliferative response to long endurance running

This randomized, double-blind, placebo-controlled study examined the influence of 6% carbohydrate ingestion on hormonal and lymphocyte proliferative responses (5 total samples over 9 hours) to 2.5 h of high-intensity running by 30 experienced marathon runners. The T-cell response differed between groups, with the placebo group exhibiting a greater increase immediately post-run and greater decrease at 3 h of recovery. No group differences were observed for Con A-, PHA-, or PWM-induced lymphocyte proliferation. However, when PHA was adjusted per T-cell, group differences were observed, highlighted by a decrease in the placebo group immediately post-run. Glucose and cortisol responses differed between groups, with glucose lower and cortisol higher in the placebo group immediately post-run. Post-run glucose correlated negatively with postrun cortisol (r=-0.670, P< 0.001) and epinephrine (r=-0.540, P=0.002). Post-run cortisol also correlated negatively with total lymphocytes and T-cells at 1.5 hours (r=-0.429, P=0.018 and r=-0.424, P=0.019, respectively) and 3 hours (r=-0.566, P=0.001 and r=-0.523, P=0.003, respectively) of recovery. The pre- to post-run change in glucose correlated to the same changes in PHA/T-cell (r=0.456, P=0.011). The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol. The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol, T-cell trafficking, and cell-adjusted PHA-induced lymphocyte proliferation following long endurance running.     

The pathologic significance of the immunoglobulins expressed by chronic lymphocytic leukemia B-cells in the development of autoimmune hemolytic anemia

Since the discovery of the CD34 stem/progenitor cell antigen, considerable progress has been made in further purifying human lymphohematopoietic stem cells (HSC). These studies have identified a number of antigens which can be targeted to subfractionate the CD34+ cell population. In particular, several lines of evidence suggest that the rare CD38(-)subpopulation of CD34+ cells may be enriched in HSC. This review briefly summarizes relevant knowledge concerning the CD38 molecule and the results of in vitro and in vivo studies of CD34+38(-)cells. Possible clinical uses for purified CD34+38(-)cells are outlined.     

The Advanced Breast Biopsy Instrumentation (ABBI), a system for stereotactic excision of mammographically suspect nonpalpable findings in the breast

Advanced Breast Biopsy Instrumentation (ABBI) allows radiologically guided stereotactic excision of non-palpable radiodense lesions with high accuracy. Tissue cylinders of 5, 10, 15 or 20 mm diameter and of variable lengths can be removed very accurately under local anaesthesia and on an outpatient basis. Thirty-six patients with suspicious clusters of microcalcifications (n = 29) and with round lesions (n = 7) of the breast were qualified for ABBI. We were able to perform the excisional biopsy in a total of 34 patients. The breast of one woman was too small to safely fit into the system and in another woman the lesion could not be visualized by the system. In 2/34 cases (6%), the excision was imprecise due to slight dislocation of the breast parenchyma by the advancing cylinder knife. In one case (3%), ABBI missed the target within a dense mastopathic breast. In all cases the excisions were well tolerated. No wound complications occurred and the cosmetic result was excellent. Histology revealed 28 benign (82%) and 6 malignant (18%) lesions. Among the 27 small microcalcifications there were 3 invasive carcinomas, 3 ductal carcinomas in situ (DCIS), 1 lobular hyperplasia, 14 mastopathies, 1 fibroadenoma, 1 duct papilloma and 4 calcifications in scars. Four of the 7 round-shaped lesions were found to be fibroadenomas, 1 lobular hyperplasia, and 2 mastopathies. With the ABBI system, non-palpable breast lesions can be precisely localized and excised.     

Effects of immobilization restraint on Syrian golden hamsters

A 2 x 2 x 4 factorial design was used to study variation of protein and fat contents in beef broths as affected by cut type (flank, shank), salt treatments (addition of salt to the medium, no salt), and initial temperatures of simmering (25, 70, 75, and 100 degrees C). Flank portions yielded slightly more protein (0.29 g/100 mL) and had three-fold less fat (0.39 g/mL) than those of shank (0.25 and 1.12 g/mL, respectively) (P < 0.05). No linear relationship of temperature and amount of extractable components was observed, but it was clear that the greatest protein extraction was accomplished when meat was immersed in cooking water at boiling point (P < 0.05). In general, salting of water reduced fat content of beef broths. However, a significant Salting x Cut type interaction showed this effect was only present in shanks (P < 0.05). Conversely, the reducing effect (P < 0.05) of salting on amount of protein extracted from flank was not observed in shanks. Based on these data, we conclude that larger amounts of protein and less fat could be transferred from meat pieces to the medium by immersing beef in salted water at the boiling point.     

A new parameter for measuring metastatic bone involvement by prostate cancer: the Bone Scan Index

In this report, we describe a method for quantitative bone scan interpretation (the Bone Scan Index or BSI) in advanced prostate cancer. The BSI estimates the fraction of the skeleton that is involved by tumor, as well as the regional distribution of the metastases in the bones. The purpose of this report is to describe the development and validation of this method in terms of reproducibility and the application of BSI for determining extent of disease and monitoring disease progression. We analyzed 263 bone scans from 90 patients being studied under four protocols at Memorial Sloan-Kettering Cancer Center for progressive, androgen-independent prostate cancer (AIPC), who had bone scans as a part of their work-up. We determined: (a) the intraobserver and interobserver variability of the BSI; (b) the comparison between a change in BSI and prostate-specific antigen (PSA); (c) the regional distribution of bony metastases in early stage D prostate cancer (<3% skeletal involvement); and (d) the rate of growth of bony metastases from prostate cancer. A cube root transformation of the percentage of involvement of the entire skeleton was used to stabilize the variance over the entire span of values (0-60% tumor involvement). The range of interobserver variability between readers was 0.2-0.5 times the cube root of the BSI (69 scans, 18 patients). Intraobserver variability was minimal when the same reader read the same scans after a 2-year interval, showing a correlation coefficient of 0.97 (reader 1) and 0.99 (reader 2), P < 0.001. There was a parallel rise in the BSI and the PSA in 24 patients (105 scans) treated for AIPC with hydrocortisone followed by suramin at PSA relapse (Pearson's moment correlation, 0.71). In a group of 27 patients with limited bone involvement by AIPC (i.e., <3% BSI), the distribution of early metastases was not random within the skeleton but was distributed in the central skeleton in a manner that matched the distribution of the normal adult bone marrow. Also, in a group of 21 patients (62 scans), the change in BSI as a function of time after diagnosis was explored graphically. The progression of bone scan changes in AIPC, from early involvement (<3%) to late involvement, was fitted to a Gompertzian equation. It showed a rapid exponential growth phase, with an estimated tumor doubling time of 43 days when the BSI was 3.3%. The change in BSI rapidly approached a more gradual slope as the percentage of skeletal involvement increased. The BSI provides a reproducible new parameter for quantitative assessment of bone involvement by AIPC. These results suggest that the BSI will be useful for stratifying patients entering treatment protocols for extent of tumor involvement of bone. Although further study is necessary, serial bone scan BSI appears capable of quantifying both the progression of bony involvement by tumor as well as the response to treatment.     

Engineering an enzyme to resist boiling

In recent years, many efforts have been made to isolate enzymes from extremophilic organisms in the hope to unravel the structural basis for hyperstability and to obtain hyperstable biocatalysts. Here we show how a moderately stable enzyme (a thermolysin-like protease from Bacillus stearothermophilus, TLP-ste) can be made hyperstable by a limited number of mutations. The mutational strategy included replacing residues in TLP-ste by residues found at equivalent positions in naturally occurring, more thermostable variants, as well as rationally designed mutations. Thus, an extremely stable 8-fold mutant enzyme was obtained that was able to function at 100 degrees C and in the presence of denaturing agents. This 8-fold mutant contained a relatively large number of mutations whose stabilizing effect is generally considered to result from a reduction of the entropy of the unfolded state ("rigidifying" mutations such as Gly --> Ala, Ala --> Pro, and the introduction of a disulfide bridge). Remarkably, whereas hyperstable enzymes isolated from natural sources often have reduced activity at low temperatures, the 8-fold mutant displayed wild-type-like activity at 37 degrees C.     

Direct association of interleukin-6 with a 130-kDa component of the interleukin-6 receptor system

Affinity cross-linking of membrane bound 125I-interleukin-6 (IL-6) on several cell lines revealed a three-band pattern of IL-6-containing cross-linked complexes with molecular masses of 100, 120, and 150 kDa. To identify the membrane components that were associated with IL-6 in the three complexes, we employed the Denny-Jaffe reagent, a heterobifunctional, cleavable cross-linker that allows the transfer of 125I from the ligand to its receptor. Samples cross-linked with Denny-Jaffe reagent were analyzed by two-dimensional SDS-polyacrylamide gel electrophoresis in which the cross-linker was cleaved prior to the second dimension. This analysis revealed that IL-6 directly associates with a 130-kDa membrane protein thus allowing the formation of the 150-kDa complex. In addition, both the 100- and 120-kDa cross-linked complexes were shown to include an 80-kDa membrane glycoprotein associated with one and two IL-6 molecules, respectively.     

Neolignans inhibit Trypanosoma cruzi infection of its triatomine insect vector, Rhodnius prolixus

Two neolignans, burchellin and nordihydroguaiaretic acid (NDGA), were toxic only to Trypanosoma cruzi clone Dm28c maintained in brain heart infusion (BHI) medium at a concentration of 100 microg/ml, not 10 microg/ml. When Rhodnius prolixus was fed with epimastigotes of T. cruzi and treated simultaneously with a single dose of burchellin or NDGA at 10 pg/ml of blood meal the number of parasites in the gut decreased. Whereas burchellin was only partially active, NDGA drastically reduced the number of epimastigotes and metacyclic trypomastigotes of T. cruzi in the excreta (urine plus feces). When the insect larvae were pretreated with burchellin or NDGA at 20 days before the infection with T. cruzi a significant reduction in the number of parasites in the gut occurred. However, when both compounds were applied at 20 days after the establishment of T. cruzi infection, although burchellin significantly reduced the gut infection, neither compound could abolish the infection entirely within the subsequent 15 days.