Increased antibiotic resistance of E. coli exposed to static magnetic fields.

The present study demonstrates that exposure of bacteria to medium strength static magnetic fields can significantly alter antibiotic sensitivity. Cultures of Escherichia coli were exposed to fields produced by permanent magnets. Samples of bacterial cultures continuously growing in the presence and in the absence of static magnetic fields were left untreated or were treated with an antibiotic and measured at 45 min intervals for cell growth and survival. It was found that exposure of E. coli to the static fields significantly increased antibiotic resistance. Bioelectromagnetics 22:129-137, 2001. Published 2001 Wiley-Liss, Inc.

Histological findings after brown recluse spider envenomation.

Histologic specimens from 41 rabbits were studied for changes resulting from the manual injection of brown recluse spider venom. Major findings included a mixed inflammatory cell infiltrate, coagulative tissue necrosis, and vasculitis. All specimens demonstrated a well-delineated zone of eosinophilic staining recognizable as "mummified" coagulative necrosis of the epidermis and dermis. A dense band of neutrophils bordered the zone of necrosis. Immediately adjacent to the neutrophilic band, small vessel vasculitis was a universal finding. Degranulated eosinophils and neutrophils and macrophages filled with eosinophilic granules were common. Inflammatory foci were often centered on groups of lipocytes within the dermis. Large vessel vasculitis resembling that seen in polyarteritis nodosa was present deep to 7 of the 40 eschars. Large vessel vasculitis may contribute to the large zones of necrosis seen after some brown recluse spider bites. Eosinophils may play a role in tissue damage after envenomation.

Drawing specimens for coagulation testing: is a second tube necessary?

Three recent studies discussed the possibility that the National Committee for Clinical Laboratory Standards (NCCLS) recommendations that the coagulation specimen should be the second or third tube collected are unnecessary. However, only one reagent/instrument was used in each study. Our protocol differed from the previous studies because we performed the assays on three different reagent/instrument systems on the same samples. Our study used photo-optic, mechanical, and nephelometric systems of clot detection. After obtaining informed consent, we obtained two blue-stoppered tubes of blood from 95 subjects: 15 normal patients and 80 patients currently on coumadin therapy. No discard tube was drawn for coagulation testing. A prothrombin time with an international normalized ratio and an activated partial thromboplastin time, were performed on each tube. Laboratory One used a MLA 1600C (Hemoliance) with Thromboplastin DS (Pacific-Hemostasis, ISI of 1.11) and APTT-LS (Pacific-Hemostasis). Laboratory Two used an STA (Diagnostica-Stago) with Neoplastine CI+ (Diagnostica-Stago, ISI of 1.14) and PTT-LT (Diagnostica-Stago). Laboratory Three used an ACL 300 with Plastinex (Biodata, ISI of 1.67) and Actin FSL (Dade Behring). No clinical or statistically significant differences were seen between the first or second tubes on any of the three reagent/instrument combinations in the PT in seconds, international normalized ratio reporting, or APTT results. Our results indicate that the NCCLS guidelines for obtaining a second tube when performing coagulation testing should be considered for elimination when new revisions are published.

Oral anticoagulant therapy and international normalized ratios in swine.

While monitoring coagulation testing in Yucatan miniature swine being given oral anticoagulants, we noticed instances of high international normalized ratios (INR) without clinical complications in our animal model. All pigs (n = 17) weighed approximately 35.2 kg and were dosed daily with 2 to 3 mg of coumadin. Plasma samples were obtained and assayed for prothrombin time (PT), calculated INR, and activated partial thromboplastin time (APTT) at baseline, and after 7 and 14 days of coumadin therapy. Results of initial testing indicated high INR values after anticoagulation and short APTT values at baseline, which led us to consider the activity of vitamin K-dependent coagulation factors in the pig. This information was not available in literature concerning this strain of swine, and was surprising given that pigs are frequently used cardiac research models. Using the same plasma samples, we repeated the PT, INR, and APTT determinations using different reagents and a different analyzer. We also determined activities of coagulation factors II, VII, IX, and X. Large PT and INR differences were seen between the two instrument/reagent combinations, possibly due to the differences in the thromboplastins used and differences in the photo-optic versus manual clot-detection method of the instruments. Vitamin K-dependent factors in all pigs responded to coumadin by decreasing to < 30.0% activity, except for factor IX. The high INR values were not as pronounced when the second instrument/reagent combination was used, and the results seemed more in line with the animals' clinical condition. With this instrument/reagent combination, the pig can be considered a good model for research requiring oral anticoagulant medication.

Peripheral blood lymphocyte apoptosis: a clue to the diagnosis of acute infectious mononucleosis.

OBJECTIVE: To describe the morphology and significance of apoptotic lymphocytes in peripheral blood smears of patients with acute infectious mononucleosis. To our knowledge this has not been previously reported. DESIGN: Peripheral blood smears from 27 patients with a positive heterophile antibody test were collected and reviewed for the presence of apoptotic lymphocytes. Flow cytometry was performed on three cases to document the previously described low expression of bcl-2 in lymphocytes in infectious mononucleosis. Four control patient populations comprising 80 cases were similarly screened for the presence of apoptotic lymphocytes. SETTING: The specimens were collected over a 3-month period in two laboratories at our tertiary care hospital; all specimens were processed according to a standard protocol. PATIENTS: Young adult military recruits and their spouses, military dependent adolescents, and retired military personnel. RESULTS: Twenty-four (88.9%) of 27 peripheral blood smears of patients with acute infectious mononucleosis contained readily identifiable apoptotic lymphocytes. Three (3.75%) of 80 control peripheral blood smears were identified with rare apoptotic lymphocytes, all occurring in patients with viral upper respiratory infections. CONCLUSIONS: The finding of apoptotic lymphocytes in a peripheral blood smear is useful in the differential diagnosis of infectious mononucleosis and neoplastic hematolymphoid processes.

Patterns of virus burden and T cell phenotype are established early and are correlated with the rate of disease progression in human immunodeficiency virus type 1-infected persons.

Human immunodeficiency virus (HIV)-1 DNA and RNA levels and T lymphocyte cell surface markers were measured in blood serum and cell fractions from asymptomatic infected patients to find novel virologic and immunologic features in early disease predictive of subsequent clinical disease course. Thirty-two patients with rapid disease progression (rapid CD4+ cell loss and progression to clinical AIDS) were compared with 25 patients with stable infections (constant or rising CD4+ cell counts, no clinical disease manifestations). All HIV-1 burdens measured by polymerase chain reaction were consistently higher in specimens from rapid progressors than slow progressors. For each patient, virus burden remained relatively constant throughout the study period (mean, 42-44 months). Flow cytometry also disclosed stable lymphocyte immunophenotype patterns that correlated strongly with subsequent rapid progression to clinical disease. Thus, in early HIV-1 infection, a constellation of high virus burden and in vivo costimulatory antigen and lymphocyte activation abnormalities is predictive of rapid disease course.

Canine epidermotropic cutaneous lymphoma.

The histologic features of an epidermotropic cutaneous lymphoma in a dog are presented. The neoplasm, which was intensely epidermotropic, thoroughly invaded the follicular and apocrine gland epithelium. The intense epidermotropism was retained in the tumor stage. By comparison to human mycosis fungoides, the cell nucleus was less convoluted and the infiltrate was less polymorphic. The lymphoid cell appeared to undergo neoplastic transformation into a cell with a large pale nucleus, often with a prominent nucleolus and variable cytoplasm. In situ marker studies are needed to clarify the cell type in this neoplasm. These studies could have relevance for the pathogenesis of human epidermotropic lymphoma.

Circadian variation of cytosol glucocorticoid receptors in human polymorphonuclear leukocytes (PMN) and mononuclear cells (MN) in a normal population.

UNLABELLED: Glucocorticoid cytosol and whole cell receptors from human PMN's have been quantified, and compared to those of human MN leukocytes on the same blood sample. The normal cytosol PMN receptor density (N = 15) averaged 1,254 +/- 105 (SE) molecules bound/cell at 0900 h and increased significantly to 1,497 +/- 98 at 2,100 h (P less than 0.02). MN cell cytosol receptor density was 1,198 +/- 145 at 0900 h and increased significantly to 1,551 +/- 117 molecules bound/cell at 2,100 h (P less than 0.01). Corresponding whole cell receptor densities at 0900 h were 2,845 +/- 273 (PMN) and 3,547 +/- 290 (MN) and these did not change significantly at 2,100 h. CONCLUSIONS: Cytosol receptors in normal human PMN and MN cells increased significantly at 2,100 h from the 0900 h level while serum cortisol levels were dropping. Whole cell receptors in the same PMN and MN cell samples did not change significantly between 0900 and 2,100 h. The normal circadian variation in serum cortisol influences the distribution of the glucocorticoid receptor between the cytosol and the nucleus, but does not influence the amount of receptor available to the whole cell. This is the first time that an endogenous physiological variation of cortisol concentration has been utilized to demonstrate a corresponding change in receptor capacity in vivo.

Cortisol secretion by an incidentally discovered nonfunctional adrenal adenoma.

We describe a middle-aged man with late-onset multiple sclerosis and an incidentally discovered asymptomatic adrenal mass. He had no symptoms or signs of hypercortisolism. A 24-h profile revealed fluctuating serum cortisol values (between 15.1 and 4.7 micrograms/dl) and inappropriately low plasma ACTH values. Urinary cortisol excretion was 89 and 106 micrograms/day on two occasions. After a 4-h ACTH infusion, serum cortisol rose from 6.3 to 108 micrograms/dl. The serum dehydroepiandrosterone level, 33 ng/dl before ACTH stimulation, did not change. During dexamethasone administration, the lowest daily urinary cortisol excretion was 37 micrograms/day, and 17-ketosteroid excretion was 8 mg/day. The response to metyrapone showed a rise of serum 11-deoxycortisol to 25.6 micrograms/dl and of ACTH to 169.5 pg/ml. After removal of the tumor, most likely an adenoma, the circadian pattern of cortisol and ACTH was normal. During a 4-h ACTH infusion, the serum cortisol level rose from 10 to 27 micrograms/dl, and dehydroepiandrosterone rose from 62 to 90 ng/dl. During dexamethasone administration, daily urinary cortisol excretion decreased to 12 micrograms/day, and 17-ketosteroid excretion dropped to 3.9 mg/day. These data show that while the tumor appeared clinically to be nonfunctional, it was producing cortisol and possibly androgens autonomously, albeit at levels too low to cause complete suppression of the pituitary-adrenal axis.

Steroid use in the elderly. Guidelines for avoiding adverse effects.

The effects of aging on adrenal function should be considered when prescribing steroid for the elderly. For glucocorticoids, the changes of decreased production and decreased clearance are relatively balanced, resulting in levels that are still within the normal range. Responses of the hypothalamic-pituitary-adrenal axis to stress are not altered. However, androgens appear to be affected by an "adrenopause," the mechanism(s) and the clinical relevance of which remain to be elucidated. The risk/benefit factors in steroid use are always of utmost concern. The physician may consider lowering the steroid dose in elderly patients of asthenic build because of the diminution of muscle mass and plasma volume that occurs with aging. Despite the physiologic changes that accompany aging, steroid used carefully and appropriately can be both safe and effective in the elderly.

Vitamin B-12 (cobalamin) deficiency: a heretofore undescribed control mechanism for plasma corticosteroid-binding globulin concentration in man.

Six of eight patients with Addisonian pernicious anemia were found to have diminished corticosteroid-binding globulin (CBG) concentrations, which returned within 2 weeks to normal after the im administration of cobalamin. T4-binding globulin was found to be completely normal before and after cobalamin replacement. Other estrogen-responsive proteins, i.e. ceruloplasmin, alpha 1-trypsin inhibitor, haptoglobin, transferrin, and alpha 2-macroglobulin, also did not follow CBG concentration in a parallel fashion. The immunoglobulins similarly did not follow a course parallel to that of CBG. The CBG concentration in two untreated patients had a normal increase in response to estrogen administration, similar to that described in CBG deficiency from other causes. No clue was found regarding the mechanism by which B12 influences the putative hepatic control of CBG concentration. It is speculated that at least two control points may be necessary for a normal CBG concentration. Patients with a genetic deficiency of CBG may also have to have cobalamin deficiency in order for them to reach a concentration approaching zero.

ACTH-induced cortisol production in multiple sclerosis.

We measured the total daily urinary cortisol produced by each of 8 patients with multiple sclerosis during intravenous therapy with adrenocorticotropic hormone (ACTH). Results indicated marked daily variation from patient to patient as well as in the same patient, suggesting that ACTH infusions do not result in reliable, consistent, high-level production of cortisol. We attempted to express the quantity of cortisol produced as an equivalent amount or oral prednisone. This comparison is difficult to make; but, given certain assumptions, the figures suggest that 40 U or 80 U ACTH infusions do not elicit the quantities of steroid generally considered to be indicated for the therapy of major allergic, autoimmune, or neoplastic diseases.

Comparison of injected and oral polio vaccine for booster immunization during the 1979 polio outbreak in Lancaster County, Pennsylvania.

During a 1979 outbreak of poliomyelitis in Lancaster County, Pennsylvania, we investigated the necessity for and the response to booster vaccination of hospital personnel. The immune response of hospital employees who received booster injections of Salk vaccine (n = 38) was compared with that of residents in the surrounding community who received Sabin trivalent OPV boosters (n = 43). Serum neutralizing antibody titers to the three polio serotypes were measured before and after booster immunization. Results indicated that 38% of the subjects in both groups had low initial antibody titers. Salk vaccine was in all circumstances as effective or better than Sabin vaccine in increasing neutralizing IgG antibody titers.

Effects of (Bu2)-cAMP and clomiphene on serum concentrations of several proteins and hormones in gonadectomized monkeys.

Sixteen mumol kg-1 of dibutyryl cyclic adenosine 3',5'-monophosphate [(Bu)2cAMP] given intramuscularly to gonadectomized green monkeys (Cercopithecus aetheops sabeus) for two-week periods, produced a significant elevation in the serum concentrations of thyroxine-binding globulin (TBG), fibrinogen, and haptoglobin. No changes were noted in cortisol-binding golbulin (CBG), cortisol, thyroxine, transferrin, total proteins, and albumin. Clomiphene citrate (12.5 mg orally) given concomitantly with (Bu)2cAMP blocked partially the (Bu)2cAMP-induced fibrinogen increment, but not the change in TBG concentration.

Surgical management of Cushing's syndrome.

Twenty-seven operations for Cushing's syndrome were reviewed. Included were five cases of adenoma and 22 of adrenal cortical hyperplasia. Preoperative laboratory data, particularly the metyrapone test, were highly accurate in distinguishing adenoma from hyperplasia. Bilateral flank incisions are preferable to the transabdominal approach, with fewer complications, less postoperative ileus, and shorter hospital stays. The inability to inspect both glands simultaneously is of little consequence because biochemical testing data are sufficiently accurate to obviate the need for gross evaluation. The use of perioperative prophylaxis seems to be of importance for preventing thromboembolism in these hypercoagulable patients. The preoperative use of adrenal cortical blocking agents has not proved to yield significant advantages.

Pituitary pigmentary hormones. Relationship of melanocyte-stimulating hormone to lipotropic hormone.

Pituitary control of pigmentation has known for more than 60 years. Since 1969, beta-melanocyte-stimulating hormone (beta-MSH) has been accepted as the main pituitary pigmentary hormone in man. Its "constant companionship" with adrenocorticotrophic hormone (ACTH) has also been repeatedly demonstrated. Current investigations challenge both of these concepts. Human beta-MSH immunoreactivity has been shown to be actually due to beta-lipotropic hormone (beta-LPH), a larger molecule that within itself contains the entire amino acid sequence of beta-MSH. Human beta-MSH does not exist in vivo; it is merely an extraction artifact formed by enzymatic degradation of beta-LPH. It would appear likely that beta-LPH, not beta-MSH, is the constant companion of ACTH.