Serologic and molecular detection of granulocytic ehrlichiosis in Rhode Island

A new indirect fluorescent-antibody (IFA) assay with antigen produced in vitro in the human promyelocytic leukemia cell line HL60 was used to identify the first recognized case of human granulocytic ehrlichiosis in Rhode Island. This IFA assay was used to detect granulocytic ehrlichiae in white-footed mice and in a dog inhabiting the area surrounding the patient's residence. Host-seeking Ixodes scapularis ticks found in the same habitat also were infected. I. scapularis ticks collected from other locations were fed on dogs and New Zealand White rabbits to assess the competency of these species as hosts of granulocytotropic Ehrlichia. Tick-induced infections of dogs were confirmed by serologic testing, tissue culture isolation, and PCR amplification, whereas several rabbits seroconverted but were PCR and culture negative. PCR amplification of the 16S rRNA gene and DNA sequencing of the PCR products or culture isolation was used to confirm granulocytic Ehrlichia infections in humans, dogs, white-footed mice, and ticks.     

The novel inotropic agent CGP 48506 alters force primarily by Ca(2+)-independent mechanisms in porcine skinned trabeculae

OBJECTIVES: The aim was to determine whether, and by what mechanism(s), a novel inotropic agent 5-methyl-6-phenyl-1,3,5,6-tetrahydro-3, 6-methano-1,5-benzodiazocine-2,4-dione (BA 41899) and its enantiomers directly alter the Ca2+ sensitivity of cardiac myofilaments. METHODS: Porcine ventricular trabeculae were permeabilised with Triton X-100. The relationship between force and pCa (-log[Ca2+]) was determined in the presence and absence of ATP. Troponin I was extracted, using vanadate, to produce unregulated maximally activated myofilaments. Force and actomyosin ATPase activity were measured simultaneously to determine tension cost (ATPase activity/tension). The effects of the (+) enantiomer (CGP 48506) on the twitch of intact muscle were demonstrated using rat papillary muscle. RESULTS: 100 microM BA 41899 had a pronounced Ca2+ sensitising effect on force production by porcine skinned cardiac fibres, increasing the pCa required for 50% maximal activation by 0.64 units, while suppressing maximum force by 18.3%. Resting tension was unaffected. These actions were primarily caused by CGP 48506 and were concentration dependent. At concentrations less than 100 microM, CGP 48506 also increased twitch amplitude in intact papillary muscles with no effect on resting tension, whereas 100 microM CGP 48506 increased resting force due to a slowing of relaxation. 100 microM CGP 48506 potentiated Ca(2+)-independent rigor tension in skinned trabeculae, indicating a Ca2+ sensitising mechanism unrelated to Ca2+ binding to troponin C. Tension cost was unaffected by 100 microM CGP 48506 over the entire range of activating Ca2+ concentrations. Suppression of maximum force by CGP 48506 was independent of both Ca2+ concentration and the regulatory troponin complex. CONCLUSIONS: Both the increase in Ca2+ sensitivity during submaximal activation and the depression of maximum force which are induced by CGP 48506 in skinned trabeculae occur at least partly through Ca(2+)-independent mechanisms.     

Mechanism of simultaneous solutes separation and concentration by size exclusion cyclic separation

The semicontinuous separation/concentration of two solutes with different molecular size by the size exclusion cyclic separation method is based on the opposite swelling responses of two gels to a temperature change. Experimental results of separation and concentration of solutes are verified qualitatively by the theoretical models based on the local equilibrium assumption. Separation of two solutes is shown by the breakthrough curves in coupled gel columns. In closed coupled columns, the volumetric space for the large molecules which are totally excluded from the gels becomes smaller, creating a concentrating effect as the gels swell when temperature changes. A mechanistic model is suggested to predict the large molecule concentration to increase to its solubility limit as cycle repeats.     

Evaluation of serum gamma-glutamyltransferase activity as a predictor of passive transfer status in crias

OBJECTIVE: To determine the relationship between serum gamma-glutamyltransferase (GGT) activity and serum IgG concentration in neonatal crias. DESIGN: Prospective observational study. ANIMALS: 21 llama and 4 alpaca crias from 0 to 5 days old. PROCEDURE: Serum GGT activity was measured, using a commercially available kit. Serum IgG concentration was determined by use of radial immunodiffusion. With a serum IgG concentration of 1,000 mg/dl (considered adequate passive transfer), specificity and sensitivity of serum GGT activity in the detection of failure of passive transfer were determined. Regression models were developed to determine the relationship between serum GGT activity and serum IgG concentration. RESULTS: Sensitivity ranged from 0.56 to 0.89, and specificity ranged from 0.88 to 0.31, depending on the value of serum GGT activity chosen as a threshold. Proportion of crias correctly classified ranged from 0.76 to 0.52. Regression models failed to demonstrate a significant relationship between serum GGT activity and serum IgG concentration. CLINICAL IMPLICATIONS: Passive transfer status in crias cannot be accurately predicted on the basis of serum GGT activity.     

Human papillomavirus investigation of patients with cervical intraepithelial neoplasia 3, some of whom progressed to invasive cancer

Unstable expansion of the CTG repeats in the 3' untranslated region encoding a member of the protein kinase family in the q13.3 band on chromosome 19 is a mutation specific for myotonic dystrophy. To examine the correlation between clinical expression and CTG trinucleotide repeat length, we carried out Southern blot analysis in a family with myotonic dystrophy. In this pedigree, the expanded CTG repeats were transmitted maternally. The mother had three female children. The mother had about 200 CTG repeats, and the number of repeats for each child was about 800, 1500 and 1600 in birth order. The mother and the patient with 800 repeats were unaware of muscle weakness or myotonia. Symptoms were present from age 3 years in the patient with 1500 repeats and from birth in the one with 1600 repeats. Although the mother menstruated regularly, the patients with 800 and 1500 repeats both menstruated irregularly, and the one with 1600 repeats has never menstruated. The age of onset and severity of the disease were correlated with the size of the expanded repeats. Endocrinological studies revealed that the basal levels of the gonadotropins, PRL and E2 were within normal range, and a pituitary response to LHRH was observed. These data suggest that the amenorrhea and menstrual irregularities were caused by a suprahypophyseal dysfunction. When expanded CTG repeats are transmitted maternally, abnormal products resulting from the metabolic disturbance in the affected mother may harm the fetus in utero. A heterozygous fetus, who has more CTG repeats, may be unable to metabolize the pathologic products sufficiently and therefore may become more severely affected. This may explain the exclusive maternal transmission of congenital myotonic dystrophy.     

Acute aortic dissection: association with patent ductus arteriosus

The Cuban recombinant vaccine against the hepatitis B virus was applied to a group of individuals over 65 from a geriatric home in Havana City. Dose of 20 ìg were administered within a scheme from 0.1 to 6 months in order to study its immunogenicity, which was evaluated at 2, 7, and 12 months after the first dose. 47.1% of protective titres and geometrical mean of 6.04 UI/L were obtained after 60 days. 100% of seroconversion (UI/L) and seroprotection (10 UI/L) and geometric mean of 136.93 UI/L were obtained when the immune response was evaluated on the 7th month. Finally, after a year, seroconversion and seroprotection were also of 100%, and the geometric mean was of 204.61 UI/L. It was proved the high immunogenic power of this vaccine, which makes it recommendable to protect this age group from the hepatitis B virus.     

Esmolol potentiates reduction of minimum alveolar isoflurane concentration by alfentanil

The beta subunit of human chorionic gonadotropin is potentially encoded by six genes, which can be categorized into two types based on a sequence change at codon 117: GCC for the type I and GAC for the type II genes. We previously showed that, whereas type I genes were exclusively expressed in normal breast tissues, expression of type II genes was associated with malignant transformation (Bellet, D., et al. Cancer Res., 57: 516-523, 1997). We designed a simple and robust test (the CG117 assay) that measures the percentage of type II over both types of chorionic gonadotropin beta mRNAs. Normal breast tissues consistently had a negative CG117 index, whereas cancer breast tissues showed indexes ranging from 0 to 100%. The prognostic significance of the CG117 index was investigated in a series of 99 unilateral invasive primary breast cancer patients with known long-term outcome (median follow-up, 9 years). The CG117 index was positive in 48 (48.5%) of the 99 tumor mRNA samples. The index was not significantly associated with standard prognostic parameters, including clinical and macroscopic tumor size, histopathological grade, and lymph node status or steroid receptor status. Patients with a positive CG117 index in primary tumor mRNA had significantly shorter metastasis-free survival (P = 0.014) and overall survival (P = 0.038) after surgery, compared to patients with a negative index. The prognostic significance of the CG117 index persisted in Cox multivariate regression analysis, both for metastasis-free survival (P = 0.008) and overall survival (P = 0.016), together with lymph node status (P = 0.027 and P = 0.009, respectively). These findings indicate that the CG117 index may contribute to the identification of high-risk breast cancer patients.