Dephosphorylation of cyclin-dependent kinases by type 2C protein phosphatases

Activating phosphorylation of cyclin-dependent protein kinases (CDKs) is necessary for their kinase activity and cell cycle progression. This phosphorylation is carried out by the Cdk-activating kinase (CAK); in contrast, little is known about the corresponding protein phosphatase. We show that type 2C protein phosphatases (PP2Cs) are responsible for this dephosphorylation of Cdc28p, the major budding yeast CDK. Two yeast PP2Cs, Ptc2p and Ptc3p, display Cdc28p phosphatase activity in vitro and in vivo, and account for approximately 90% of Cdc28p phosphatase activity in yeast extracts. Overexpression of PTC2 or PTC3 results in synthetic lethality in strains temperature-sensitive for yeast CAK1, and disruptions of PTC2 and PTC3 suppress the growth defect of a cak1 mutant. Furthermore, PP2C-like enzymes are the predominant phosphatases toward human Cdk2 in HeLa cell extracts, indicating that the substrate specificity of PP2Cs toward CDKs is evolutionarily conserved.     

Urological needs assessment for primary care practice: implications for undergraduate medical education

PURPOSE: The majority of graduating medical students will become primary care physicians. We discuss what urological topics primary care physicians need to know, proficiency in urological skills of primary care physicians and undergraduate urological curriculum recommendations that would serve the needs of primary care physicians. MATERIALS AND METHODS: A written survey instrument was mailed to all 452 family practice residency directors in the United States. Subjects rated how well family practice physicians need to know urological topics and how proficient they must be in urological skills. RESULTS: A total of 329 responses were received (response rate 73%). Respondents indicated that they need to know urinary tract infection, sexually transmitted disease, epididymitis, benign prostatic hyperplasia diagnosis, hematuria and erectile dysfunction diagnosis very well. They indicated a greater need to know diagnostics than staging or management of prostate, bladder, renal or testis cancer (p <0.05). Among diagnostics respondents indicated that generalists should be highly proficient in urinalysis interpretation. Proficiency was ranked greater for interpretations of semen analysis than excretory urogram, abdominal computerized tomography or magnetic resonance imaging, or renal ultrasound (p <0.05). Proficiency in catheter insertion and circumcision was ranked highly, and vasectomy was ranked moderately. CONCLUSIONS: Primary care physicians need to know and be proficient in many urological topics and skills. They need to know diagnostics better than staging and management information, and urological infectious topics better than urological cancers. They need to be proficient in physical examination, urinalysis interpretation, catheter insertion, circumcision and vasectomy. Undergraduate medical education should reflect these needs.     

Identification of a pathogenicity island required for Salmonella survival in host cells.

We have identified a region unique to the Salmonella typhimurium chromosome that is essential for virulence in mice. This region harbors at least three genes: two (spiA and spiB) encode products that are similar to proteins found in type III secretion systems, and a third (spiR) encodes a putative regulator. A strain with a mutation in spiA was unable to survive within macrophages but displayed wild-type levels of epithelial cell invasion. The culture supernatants of the spi mutants lacked a modified form of flagellin, which was present in the supernatant of the wild-type strain. This suggests that the Spi secretory apparatus exports a protease, or a protein that can alter the activity of a secreted protease. The "pathogenicity island" harboring the spi genes may encode the virulence determinants that set Salmonella apart from other enteric pathogens.     

Ultrasound Densitometric Analysis: Comparison Between an Online Digital Acquisition Acoustic Program and an Offline Analog Program

Videodensitometric analysis of myocardial contrast echocardiography is traditionally performed off line. Recently, an online contrast ultrasound analysis system, Acoustic Densitometry (Hewlett-Packard), was introduced. We compared pixel intensities acquired with Acoustic Densitometry to pixel intensities derived from videodensitometry. A tissue phantom was imaged in phase I using three transducer frequencies (2.5, 3.5, and 5.0 MHz). In phase II, an in vitro flowing tube model with various concentrations of Albunex® was imaged at two flow rates, 0.6 and 1.2 m/sec, and at two transducer frequencies, 2.5 and 3.5 MHz. The relationship between pixel intensities yielded by the two systems for identical ultrasound signals was determined with linear regression. Intensities derived with Acoustic Densitometry strongly correlated with those derived from the offline videodensitometry system. The intensities were related by a predictive multiplicative factor based on display characteristics of the two systems. These results suggest that semiquantitative, online perfusion analysis with Acoustic Densitometry is as sensitive as analysis offline with videodensitometry.     

Unintentional intraoperative awareness during sufentanil anaesthesia for cardiac surgery

Purpose

The aim of this clinical report is to describe a case of unintentional intraoperative awareness during sufentanil anaesthesia in a patient undergoing elective aortocoronary bypass grafting.

Clinical findings

After premedication with morphine (5 mg) and scopolamine (0.2 mg), this 51-yr-old woman received sufentanil (10 μg · kg^?1), midazolam (4 mg) and isoflurane (0.3–0.4% end-tidal). The patient recalled specific events and discussions which took place in the operating room during surgery. This patient’s report was clear and corroborated by operating room personnel. The patient denied having felt pain, anxiety or emotional distress.

Conclusion

Although awareness during opioid anaesthesia has been previously described with morphine and fentanyl, as far as we know this is the first clinical report of awareness with sufentanil. Given that recent efforts of early extubation in cardiac surgery patients may involve a reduction in the amount of opioid administered, this report serves as a reminder of the ever present potential for this disturbing complication.     

Duodenal ulcer and working-class mobility in an African population in South Africa

The number of Africans in Johannesburg presenting with duodenal ulcers has steadily increased over the past 50 years. The characteristics of 105 patients with duodenal ulcer who presented a Baragwanath Hospital were compared with those of matched and unmatched samples of patients without gastrointestinal conditions in the same hospital. Men with duodenal ulcers were found to be significantly better educated than their controls, most had been born in the town, and more of them were employed at higher, though not the highest, educational levels. These data were used to test Susser's proposition that duodenal ulcers are associated with "early urbanisation." Johannesburg blacks with duodenal ulcer did seem to fit the pattern, but the relation between stress and duodenal ulcer remains unclear.     

Cyclic nucleotide phosphodiesterase. Insulin activation detected in adipose tissue by gel electrophoresis.

In rat adipose tissue, insulin (100 muU./ml.) increases the activity of cyclic AMP (but not cyclic GMP) phosphodiesterase (PDE). Radioisotopic assay, autoradiography, and histochemical stains demonstrated that cyclic nucleotide PDE activity is associated with multiple bands of protein separable by polyacrylamide gel electrophoresis. The insulin activation of cyclic AMP PDE, however, was limited to a single band corresponding to the "low" Km enzyme specific for cyclic AMP; insulin had no effect on the "high" Km enzyme or on PDE bands with activity directed toward cyclic GMP. These data support the concept that insulin may modulate the activity of at least one of the cyclic AMP phosphodiesterases.