Interaction of substrate and effector binding sites in the ArsA ATPase

The ars operon of plasmid R773 confers resistance to antimonials and arsenicals in Escherichia coli by encoding an ATP-dependent extrusion system for the oxyanions. The catalytic subunit, the ArsA protein, is an ATPase with two nucleotide binding consensus sequences, one in the N-terminal half and one in the C-terminal half of the protein. The ArsA ATPase is allosterically activated by tricoordinate binding of As(3+) or Sb(3+) to three cysteine thiolates. Previous measurements suggested that the intrinsic fluorescence of tryptophans might be useful for examining binding of Mg2+ ATP and antimonite. In the present study an increase in intrinsic tryptophan fluorescence was observed upon addition of Mg2+ ATP. This enhancement was reversed by addition of antimonite. The ArsA protein contains four tryptophan residues: Trp159, Trp253, Trp522, and Trp524. The first two were altered to tyrosine residues by site-directed mutagenesis. Cells expressing both the arsAW159Y and arsAW253Y mutations retained resistance to arsenite, and the purified W159Y and W253Y proteins retained ATPase activity. While the intrinsic tryptophan fluorescence of the W253Y protein responded to addition of Mg2+ ATP, intrinsic tryptophan fluorescence in the purified W159Y protein was no longer enhanced by substrate. These results suggest that Trp159 is conformationally coupled to one or both of the nucleotide binding sites and provides a useful probe for the interaction of effector and substrate binding sites.     

Why clinicians infrequently do research.

Findings from several studies have indicated that the scientist-practitioner model (which stresses the integration of research into clinical practice) is infrequently adopted by practicing clinical psychologists. Although many hypotheses have been offered, there have been no empirical examinations of the reasons why clinicians infrequently engage in research. In this study, 79 practicing clinical psychologists completed a questionnaire focusing on perceived impediments to clinical research. A number of inhibiting factors were identified, but the intrinsic and extrinsic contingencies associated with the conduct of research were most salient. Other concerns included ethics, the impact of research on the clinical process, and the unavailability of appropriate research designs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Radiological case of the month. Reflex sympathetic dystrophy

The authors report a case of pregnancy in a rudimentary horn diagnosed by laparotomy. The location could not be suspected by any clinical sign. This pregnancy stopped at 19 weeks of amenorrhoea. Any attempts of induction of labour failed. Laparotomy was necessary to assess the diagnosis and to allow the resection of the uterine horn. The authors present a review of literature.     

Interleukin-1 beta-converting enzyme mediates cisplatin-induced apoptosis in malignant glioma cells

Increasing the susceptibility of tumor cells to apoptotic cell death following chemotherapy is of importance to the outcome of cancer treatment. Although the tumor suppressor gene p53 is required for efficient induction of apoptosis by chemotherapeutic agents, it is not the only apoptosis mediator gene. The molecular mechanisms mediating apoptosis following chemotherapy via p53-dependent or p53-independent pathways remain unclear. We show here that cis-diamminedichloroplatinum (cisplatin) induces the expression of interleukin-1 beta-converting enzyme (ICE), a mammalian homologue of the Caenorhabditis elegans cell death gene ced-3, in murine and human malignant glioma cells during apoptosis regardless of their p53 status. Furthermore, overexpression of the murine ICE gene induces apoptosis in these tumor cells. The apoptosis induced by cisplatin treatment or murine ICE overexpression can be suppressed by the tetrapeptide ICE inhibitor Ac-YVAD-CMK or the apoptosis inhibitors bcl-2 or bcl-2-related bcl-XL gene. These findings suggest that ICE may mediate apoptosis induced by chemotherapy, and its induction could represent a novel approach for the effective treatment of malignant glioma.     

Placebo-controlled immunotherapy with Cocos nucifera pollen extract

The effectiveness of Cocos nucifera pollen extract immunotherapy (CPE-IT) was studied in 96 patients allergic to C. nucifera pollen. A placebo-controlled study was performed at random for a period of 6-12 months. The clinical status of the patients measured by the symptom-medication scores demonstrated that C. nucifera pollen-allergic patients had significant (p < 0.005) clinical improvement after CPE-IT in comparison to placebo treatment. Serological study resulted a significant reduction (p < 0.001) of specific IgE and significant elevation (p < 0.01) of specific IgG in post-therapeutic patients' sera which were correlated significantly (r = 0.45, p < 0.001); the changes of the above immunoglobulin levels in the placebo-treated patients were nonsignificant. However, there was no correlation between symptom-medication scores and changes in specific serum IgE or IgG levels.     

The use of external, noninvasive pacing for the termination of supraventricular tachycardia in the emergency department setting

STUDY OBJECTIVE: To determine the potential feasibility of external cardiac pacing for the termination of sustained supraventricular tachycardia in the emergency department setting. TYPE OF PARTICIPANTS: Three men and two women (mean age, 34 years) who presented to the ED with a narrow-complex, hemodynamically stable tachycardia that was later proven to be supraventricular in origin. INTERVENTION: Each patient underwent external overdrive pacing using a modified external pacemaker at a pulse amplitude of 120 mA and a rate between 240 and 280 pulses per minute. RESULTS: In four patients, external cardiac pacing was able to successfully terminate the tachycardia without complication. In one patient, the pacemaker was not able to terminate the tachycardia. CONCLUSION: We conclude that external, noninvasive pacing is a feasible means of terminating supraventricular tachycardia in the ED setting.     

Cryptosporidium parvum initiates inflammatory bowel disease in germfree T cell receptor-alpha-deficient mice

Flora-bearing mice with targeted disruption of T cell receptor (TCR)-alpha or -beta genes spontaneously develop intestinal inflammation with features similar to ulcerative colitis in humans. TCR-alpha-deficient mice maintained germfree or colonized with a limited number of intestinal bacteria failed to develop inflammatory bowel disease (IBD)-like lesions. Evidently, inflammation in these mice does not develop spontaneously or result from a generalized antigenic stimulation, but rather requires induction by a heretofore unidentified specific stimulus. We describe the development of IBD-like lesions in germfree TCR-alpha-deficient mice monoassociated with the protozoan Cryptosporidium parvum. Lesions were seen in distal ileum, cecum, and colon and were most severe in the cecum. A prominent leukocytic infiltrate within the lamina propria was a common characteristic of the lesions observed in the C. parvum-infected germfree TCR-alpha-deficient mice. The leukocytic infiltrate was composed of aggregates of B220+ cells, the majority of which expressed surface IgD (ie, conventional B lymphocytes). It has been proposed that antigenic stimulation by a microorganism(s) is needed to initiate intestinal inflammation in TCR-alpha-deficient mice. Our results indicate that a single microbial species, C. parvum, is capable of triggering the development of IBD-like lesions in germfree TCR-alpha-deficient mice.     

Infant affect and affect regulation during the still-face paradigm with mothers and fathers: the role of infant characteristics and parental sensitivity

This laboratory study examined mothers' and fathers' sensitivity during face-to-face interactions with their infants as well as infants' affective and regulatory responses during mother-infant versus father-infant still face (SF). The degree to which infant gender and temperament as well as parental sensitivity predicted SF responses was also examined. Participants included 94 healthy, primarily White, middle-class 4-month-olds and their parents. Results indicated that mothers and fathers were equally sensitive toward their infants. Infants' affect and regulatory behaviors were also significantly stable across mother- and father-infant SF situations, although several differences in mean levels of regulation emerged. Finally, the extent to which exogenous and endogenous variables predicted infant SF responses differed as a function of which affect or regulatory variable was being examined and with which parent the infant was experiencing SF.