Improvements in the infectivity of cryopreserved third-stage larvae of Angiostrongylus cantonensis using a programmable freezer.

Although there have been some advances in the cryopreservation of Angiostrongylus cantonensis, the degrees of viability and infectivity of the cryopreserved developmental stages have not been high. A two-step freezing protocol using a programmable freezer was determined to be effective in improving the infectivity of the cryopreserved third-stage larvae of this parasite. After washing steps and suspension in 10% (v/v) dimethylsulfoxide and equilibrium at room temperature the larvae were placed into the freezer.

Weaning from mechanical ventilation.

For the ventilator-dependent patient, weaning should be accomplished by withdrawing support safely, efficaciously, and efficiently. Success depends largely on physiologic determinants of respiratory system function, avoidance of ventilator-associated complications, and attention to patient readiness. Recent clinical trials, predictors of weaning, current techniques of weaning, the concept of reloading the respiratory pump, and determinants of ventilator dependency are all discussed.

Muscular dystrophy in mdx mice despite lack of neuronal nitric oxide synthase.

Neuronal nitric oxide synthase (nNOS) is a component of the dystrophin complex in skeletal muscle. The absence of dystrophin protein in Duchenne muscular dystrophy and in mdx mouse causes a redistribution of nNOS from the plasma membrane to the cytosol in muscle cells. Aberrant nNOS activity in the cytosol can induce free radical oxidation, which is toxic to myofibers.

Seven novel mammalian SNARE proteins localize to distinct membrane compartments.

Soluble N-ethylmaleimide-sensitive factor-attachment protein receptor (SNARE) proteins of the vesicle-associated membrane protein (VAMP) and syntaxin families play a central role in vesicular trafficking through the formation of complexes between proteins present on vesicle and target membranes. Formation of these complexes is proposed to mediate aspects of the specificity of vesicle trafficking and to promote fusion of the lipid bilayers. In order to further understand the molecular mechanisms that organize membrane compartments, we have characterized seven new mammalian proteins of the VAMP and syntaxin families.

BCL-2 family: regulators of cell death.

An expanding family of BCL-2 related proteins share homology, clustered within four conserved regions, namely BCL-2 homology (BH1-4) domains, which control the ability of these proteins to dimerize and function as regulators of apoptosis. Moreover, BCL-XL, BCL-2, and BAX can form ion-conductive pores in artificial membranes. The BCL-2 family, comprised of both pro-apoptotic and anti-apoptotic members, acts as a checkpoint upstream of CASPASES and mitochondrial dysfunction.

Molecular characterization of Gluconobacter oxydans recA gene and its inhibitory effect on the function of the host wild-type recA gene.

A DNA fragment containing the recA gene of Gluconobacter oxydans was isolated and further characterized for its nucleotide sequence and ability to functionally complement various recA mutations. When expressed in an Escherichia coli recA host, the G. oxydans recA protein could efficiently function in homologous recombination and DNA damage repair.

Patient-ventilator trigger asynchrony in prolonged mechanical ventilation.

Study Objective: To investigate patient-ventilator trigger asynchrony (TA), its prevalence, physiologic basis, and clinical implications in patients requiring prolonged mechanical ventilation (PMV).

Study Design: Descriptive and prospective cohort study.

Setting: Barlow Respiratory Hospital (BRH), a regional weaning center.

A strategy for rapid, high-confidence protein identification.

A procedure is described for rapid, high-confidence identification of proteins using matrix-assisted laser desorption/ionization tandem ion trap mass spectrometry in conjunction with a genome database searching strategy. The procedure involves excision of copper-stained bands or spots from electrophoretic gels, in-gel trypsin digestion of the proteins, single-stage mass spectrometric analysis of the resultant mixture of tryptic peptides, followed by tandem ion trap mass spectrometric analysis of selected individual peptides, and database searching of the relevant genomic database using the program PepFrag. The scheme provides sensitive, real-time protein identification as well as facile identification of modifications.

Update and review of blastomycosis.

Blastomycosis has a wide spectrum of clinical presentations. When a patient presents with chronic pneumonia, especially coexisting with cutaneous lesions, blastomycosis infection needs to be considered in the differential diagnosis. Erythema nodosum can rarely be associated with pulmonary blastomycosis.

BCL-XL-regulated apoptosis in T cell development.

Thymocyte differentiation progresses through well-defined stages in which apoptosis is central to the selection of a functional TCR repertoire. In the present study, we explored the developmental effects of BCL-XL, a repressor of apoptosis. We found that endogenous BCL-XL is down-regulated by both positive and negative selection signals at the CD4+ CD8+ stage of thymocyte development.

A phase II study of continuous infusion 5-fluorouracil (5-FU) with epirubicin and cisplatin in metastatic, hormone-resistant prostate cancer: an active new regimen.

The aim of this phase II study was to examine the efficacy of combination chemotherapy comprising epirubicin, cisplatin and protracted infusion 5-fluorouracil in patients with metastatic prostate cancer. 21 patients were treated, of whom 9 (43%) responded for a minimum of 7 months. Continuation of previously effective second-line hormone therapies appeared to be a determinant of response; only 1 of 6 patients responded after its discontinuation.

Post-ICU mechanical ventilation: treatment of 1,123 patients at a regional weaning center.

Study Objectives: To update our database, reporting changes in the results of weaning attempts and profile of patients transferred to us after prolonged mechanical ventilation (PMV) in the ICU.

Design: Retrospective record review, with prospective recording of physiologic measurements on admission from mid-1994.

Setting: Regional weaning center (RWC).

BRCA1 is a component of the RNA polymerase II holoenzyme.

The familial breast-ovarian tumor suppressor gene product BRCA1 was found to be a component of the RNA polymerase II holoenzyme by several criteria. BRCA1 was found to copurify with the holoenzyme over multiple chromatographic steps. Other tested transcription activators that could potentially contact the holoenzyme were not stably associated with the holoenzyme as determined by copurification.

Two-dimensional electrophoretic analyses of urinary proteins in Chinese male urinary stone patients.

This study was aimed to detect the differences in soluble urinary proteins between normal Chinese male individuals and urinary stone formers by 2-dimensional electrophoresis. Twenty urine samples were obtained from normal male adults and 35 from recurrent male urinary stone formers. The stone compositions included calcium oxalate, uric acid, carbonate apatite, and brushite.

Protein interactions regulating vesicle transport between the endoplasmic reticulum and Golgi apparatus in mammalian cells.

The proposed cis-Golgi vesicle receptor syntaxin 5 was found in a complex with Golgi-associated SNARE of 28 kDa (GOS-28), rbet1, rsly1, and two novel proteins characterized herein: rat sec22b and membrin, both cytoplasmically oriented integral membrane proteins. The complex appears to recapitulate vesicle docking interactions of proteins originating from distinct compartments, since syntaxin 5, rbet1, and GOS-28 localize to Golgi membranes, whereas mouse sec22b and membrin accumulate in the endoplasmic reticulum. Protein interactions in the complex are dramatically rearranged by N-ethylmaleimide-sensitive factor.

Functional antagonism between RNA polymerase II holoenzyme and global negative regulator NC2 in vivo.

Activation of eukaryotic class II gene expression involves the formation of a transcription initiation complex that includes RNA polymerase II, general transcription factors, and SRB components of the holoenzyme. Negative regulators of transcription have been described, but it is not clear whether any are general repressors of class II genes in vivo. We reasoned that defects in truly global negative regulators should compensate for deficiencies in SRB4 because SRB4 plays a positive role in holoenzyme function.

Nitric oxide synthase and cyclic GMP-dependent protein kinase concentrated at the neuromuscular endplate.

Nitric oxide mediates diverse functions in development and physiology of vertebrate skeletal muscle. Neuronal type nitric oxide synthase-mu is enriched in fast-twitch fibers and binds to syntrophin, a component of the sarcolemmal dystrophin glycoprotein complex. Here, we show that cyclic GMP-dependent protein kinase type I, a primary effector for nitric oxide, occurs selectively at the neuromuscular junction, in mice and rats, and both neuronal type nitric oxide synthase-mu and cyclic GMP-dependent protein kinase type I remain at skeletal muscle endplates at least two weeks following muscle denervation.

The trouble with family medicine.

Background: The trouble with family medicine is that the perceptual framework it uses to view the phenomena of health and illness is at variance with the frameworks traditionally used by medicine generally. This creates difficulties in communication between those in family medicine and those in other disciplines, and sometimes leads to misunderstanding of the nature of the discipline of family medicine and its place in the health care system. Those who practise family medicine need to be 'multilingual', able to understand and speak the language and use the metaphors of family medicine, yet equally able to use the language and metaphors of other disciplines.

Two theoretical and practical aspects of knitting technique: invariants and a new class of Petri net.

We present two aspects of knitting technique, the structural properties (especially the P- and T-invariants), and the synchronized choice net (a new class of Petri net), that are of both theoretical importance and practical uses to the verification of structural correctness of a Petri net or to detect the structural problem of a Petri net. This work first proves that the ordinary Petri nets synthesized with knitting technique are structurally bounded, consistent, conservative and safe (when each home place holds one token) using the well-known linear algebra approach. It also provides a procedure for finding P- and T-invariants for Petri net synthesized using the knitting technique.

Impact of renal dysfunction on weaning from prolonged mechanical ventilation.

BACKGROUND: In the intensive care unit (ICU) setting, the combination of mechanical ventilation and renal replacement therapy (RRT) has been associated with prolonged length of hospital stay, high cost of care and poor outcome. We gathered outcome data on patients who had severe renal dysfunction on transfer to our regional weaning center (RWC) for attempted weaning from prolonged mechanical ventilation (PMV). We screened the admission laboratory values of 1077 patients transferred to our RWC over an 8-year period.

Regulation of neuronal nitric oxide synthase through alternative transcripts.

Nitric oxide (NO) participates in diverse physiological processes ranging from neurotransmission to muscle relaxation. Neuronal-derived NO can be either beneficial or detrimental depending on the cellular context. Neuronal NO synthase (nNOS) must therefore be tightly regulated.

Supernatant from cultured intestinal cells inhibits the growth of gram-negative bacteria.

Bacterial growth chambers of Transwell units bearing intestinal epithelial monolayers (C2BBe) was consistently observed to be stagnant during the course of transmigration studies with Salmonella typhi. This limitation could not be explained by varying the bacterial load in the inoculum. Conditioned media produced by cultured C2BBe cells would not support bacterial growth.

The importance of continued endocrine treatment during chemotherapy of hormone-refractory prostate cancer.

Objective: It is unclear if continuing with androgen deprivation in hormone-refractory prostate cancer is of any benefit. We report here 3 cases where continuation of androgen deprivation was essential in maintaining the response to chemotherapy.

Results: In the 3 patients there was an initial response to chemotherapy before relapse, as assessed by prostate-specific antigen levels.

BAX-induced cell death may not require interleukin 1 beta-converting enzyme-like proteases.

Expression of BAX, without another death stimulus, proved sufficient to induce a common pathway of apoptosis. This included the activation of interleukin 1 beta-converting enzyme (ICE)-like proteases with cleavage of the endogenous substrates poly(ADP ribose) polymerase and D4-GDI (GDP dissociation inhibitor for the rho family), as well as the fluorogenic peptide acetyl-Asp-Glu-Val-Asp-aminotrifluoromethylcoumarin (DEVD-AFC). The inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD-fmk) successfully blocked this protease activity and prevented FAS-induced death but not BAX-induced death.