Identification of myoglobin in human smooth muscle

Myoglobin (Mb) has been believed to be absent generally from mammalian smooth muscle tissue. Examination of human rectal, uterine, bladder, colon, small intestine, arterial, and venous smooth muscle by immunohistochemical techniques shows that each of these tissues is immunopositive for both smooth muscle myosin and human Mb. Mb-specific primers were used for the polymerase chain reaction to generate cDNA from smooth muscle tissues. Southern hybridization with a Mb-specific probe gave a very strong signal with the cDNA from rectum, weaker signals from small intestine and uterus, a faint signal from colon, and no signal from bladder tissue. High performance liquid chromatography analysis coupled with sequence determination has shown that contaminating heme-binding serum albumin as well as hemoglobin in extracts of smooth muscle seriously compromise any heme-based or spectrophotometric assay of Mb. Combined affinity and size exclusion chromatography, however, provide the necessary resolution. The cDNA-derived amino acid sequence of human smooth muscle Mb was found to be identical to that of Mb from striated muscle.     

Toxoplasmosis in goats in the Netherlands: a pilot study

A pilot-study was carried out on ten Dutch goat farms to see whether there is a relationship between farm management factors and the occurrence of toxoplasmosis. Questionnaires were used to collect information about farm management factors and blood samples were taken to determine the prevalence of toxoplasmosis on these farms. The mean prevalence was 47% (range 5-90%). The presence of kittens on a farm was a risk factor for a higher prevalence of toxoplasmosis.     

A telescope-design directional neutron spectrometer

A directional spectrometer that uses a superheated emulsion of dichlorotetrafluoroethane at the centre of a 30 cm diameter moderating-sphere of nylon-6. The system has a telescope-design wherein the detector views a narrow solid angle of about 1/6 steradians. The hydrogenous sphere effectively attenuates laterally incident neutrons, thus providing a strong angular dependence of the response. The central detector is sequentially operated at seven temperatures between 25 and 55 degrees C in order to generate a matrix of nested response functions suitable for few-channel spectrometry. The response matrix of the system has been determined by calibrations with monoenergetic neutrons and by Monte Carlo neutron transport calculations. The double-differential unfolding method developed for this system applies the principle of maximum entropy and allows for the rigorous use of all a priori information. The spectrometer is intended for use in the mixed neutron/photon fields encountered in the nuclear power industry, being suitable for spatially distributed radiation sources with maximum neutron energies up to 10 MeV.     

Diaphragm activation with intramuscular stimulation in dogs

We studied in 10 supine anesthetized dogs diaphragm contraction produced by electrical activation with intramuscular electrodes surgically implanted in the ventral surface of the diaphragm and compared this with activation of the ipsilateral phrenic nerve (C5, 6, and 7) before it entered the thorax. Repetitive 40-Hz pulse trains with supramaximal current stimulus were used after hyperventilation of the animals to apnea. A single intramuscular electrode within 1 to 2 cm of the site of phrenic nerve entry into the diaphragm produced a mean transdiaphragmatic pressure of 12.0 cm H2O +/- 0.97 SE and mean tidal volume of 0.27 L +/- 0.04 SE. Mean values observed with phrenic nerve stimulation were not statistically different, and both electrode systems produced equivalent outward abdominal motion and upper rib cage paradox, as monitored by inductive plethysmography. There was no difference in gas exchange during stimulation with a single hemidiaphragm electrode and mechanical ventilation compared at the same tidal volume and respiratory rate. Blockade of neuromuscular transmission with curare eliminated intramuscular and phrenic nerve stimulation proportionately, suggesting that activation of the diaphragm is dependent in both cases on the phrenic nerve. This technique does not entail manipulation of the phrenic nerve and may have clinical application as an alternative technique for diaphragm pacing.     

Structural features determining the antibiotic potencies of natural and synthetic hop bitter resins, their precursors and derivatives

Twenty-six hop bitter resins, some hitherto not investigated, were tested for antimicrobial activities. Gram-positive bacteria were much more sensitive than Gram-negative ones. The inhibitory effect against Bacillus subtilis 168 was measured by several methods and the general rule could be established that the antibiotic properties are mainly dependent on the hydrophobic parts of the molecules. Thus the acyl-lupuphenones (2-acyl-3,5-4,4',6-tri(3-methyl-2-butenyl)-cyclohexane-triones (1, 3, 5) having three prenyl and one acyl side chain are the most active substances. Their minimum inhibitory concentration (MIC) increases from the capro (0.5 muM) to the aceto derivative (11 muM). Any substitution with hydrophilic functions or loss of hydrophobic groups causes reductions in biological activity. This is most evident with the corresponding acyl-phloroglucine precursors (2-acyl-1,3,5-trihydroxybenzenes) which lack the three prenyl side chains (MIC, 110 to 5050 muM respectively). Conversion of the central six-membered ring structure into a five-membered one results in additional losses of antimicrobial activity. These findings support the proposal that the lipophilic region of the cell membrane represents the target site for the hop bitter resins.     

A technique for identifying the subgroup membership of certain misarticulating children

Cluster analysis was used to identify two homogeneous clusters of 8-9 1/2-year-old children who misarticulated /s/, /r/, or both. The analysis was based on the children's scores on 40 measures of language, reading, auditory processing, and other variables. Discriminant function analysis was then used to identify a subset of five measures and a means of computing classification scores. These measures and the classification scores can be used to identify the cluster membership of new subjects. The use of classification scores for identifying cluster membership was cross-validated against cluster analysis of a second group of children. The two clusters are described in terms of their performance on language and reading measures.     

Amifostine protects normal tissues from paclitaxel toxicity while cytotoxicity against tumour cells is maintained

The objectives of this study were to evaluate the protective effects of amifostine against paclitaxel-induced toxicity to normal and malignant human tissues. Haematopoietic progenitor colony assays were used to establish the number of CFU-GEMM and BFU-E colonies after incubation with WR-1065 alone, Amifostine alone, paclitaxel (2.5 or 5 microM) +/- WR-1065 or amifostine. MTT and alkaline elution assays evaluated the in vitro growth inhibitory and DNA damaging effects, respectively, of paclitaxel with or without amifostine against normal human fibroblasts and human non-small cell lung cancer (NSCLC) cells. This combination was also evaluated in vivo using severe combined immune deficient (scid) mouse models of early (non-palpable tumours) and advanced (palpable tumours) human ovarian cancer. Human 2780 ovarian cancer cells were inoculated subcutaneously while paclitaxel and amifostine were administered intraperitoneally. A brief exposure (15 min) to amifostine not only protected human haematopoietic progenitor colonies from paclitaxel toxicity, but stimulated the growth of CFU-GEMM and BFU-E beyond control values. Amifostine protected normal human lung fibroblasts from paclitaxel-induced cytotoxicity and DNA single-strand breaks. However, paclitaxel cytotoxicity and DNA single-strand breaks were actually enhanced by pretreatment with amifostine in the NSCLC model. Importantly, amifostine did not interfere with paclitaxel antitumour activity even with prolonged exposure (24.5 h) of the lung cancer cells to high concentrations (1.2 mM) in vitro or following five repetitive high doses (200 mg/kg) given to scid mice with human ovarian cancer xenografts. Indeed, under certain circumstances, amifostine resulted in sensitisation of tumour cells to paclitaxel. Our results confirm previous reports of the ability of amifostine to protect normal tissues from the toxic effects of chemotherapy drugs and now extend these observations to paclitaxel.     

Personalities and alimentary behaviors in obese patients

The ability of lonidamine (LND), an energolytic derivative of indazole-carboxylic acid, to modulate the cytotoxic activity of cisplatin (CDDP) and epidoxorubicin (EPI), singly or in combination, was investigated in two human breast cancer cell lines (MCF7 and T47D). A 72-hr post-incubation with a non-cytotoxic concentration of LND (75 microM) increased the activity of a 1-hr CDDP treatment as well as that of a 1 to 16-hr EPI treatment. A different pattern of interaction among the drugs and modulator was observed as a function of the sequence of drug treatment. Specifically, supra-additive or additive effects of the combination were obtained in the two cell lines according to the different treatment schemes. In particular, the maximum potentiation was observed in MCF7 cells simultaneously exposed to CDDP, EPI and LND for 1 hr and then post-incubated with LND for 72 hr, and in T47 first exposed to EPI and LND, then to CDDP and LND, and finally post-incubated with LND. Flow cytometric analysis of MCF7 cell distribution in the different cycle phases showed that combined treatment with EPI/CDDP/LND was able to stabilize cell cycle perturbations (mainly G2M accumulation) induced by individual agents. The ability of LND to potentiate CDDP and EPI cytotoxicity, and the consideration that LND causes side effects different from those caused by alkylating agents and anthracyclines, make this compound an attractive candidate for multidrug combination therapy in breast cancer.