Identification and characterization of a novel 9.2-kDa membrane sector-associated protein of vacuolar proton-ATPase from chromaffin granules

Vacuolar proton-translocating ATPase (holoATPase and free membrane sector) was isolated from bovine chromaffin granules by blue native polyacrylamide gel electrophoresis. A 5-fold excess of membrane sector over holoenzyme was determined in isolated chromaffin granule membranes. M9.2, a novel extremely hydrophobic 9.2-kDa protein comprising 80 amino acids, was detected in the membrane sector. It shows sequence and structural similarity to Vma21p, a yeast protein required for assembly of vacuolar ATPase. A second membrane sector-associated protein (M8-9) was identified and characterized by amino-terminal protein sequencing.     

An introduction to forensic dentistry

Dental science has much to offer law enforcement agencies in the detection and solution of crime. The permanent teeth develop throughout the first two decades of life, and physiologic variations, pathoses, and the effects of dental therapy may be recorded in the hard tissues of the remaining dentition throughout life and beyond. It is the role of the forensic dentist to extract this information and use it in the identification of the unknown body. The teeth may also be used as weapons and, under certain circumstances, may leave information as to the identity of the biter. Analysis of bite marks is the second major responsibility of the forensic dentist. The general practitioner has a major role to play in providing the accurate dental records on which much of forensic activity is based.     

Characteristics of human fetal spinal cord grafts in the adult rat spinal cord: influences of lesion and grafting conditions

The present study evaluated the growth potential and differentiation of human fetal spinal cord (FSC) tissue in the injured adult rat spinal cord under different lesion and grafting conditions. Donor tissue at 6-9 weeks of gestational age was obtained through elective abortions and transplanted either immediately into acute resection (solid grafts) or into chronic contusion (suspension and solid grafts) lesions (i.e., 14-40 days after injury) in the thoracic spinal cord. The xenografts were then examined either histologically in plastic sections or immunocytochemically 1-3 months postgrafting. Intraspinal grafts in acute lesions demonstrated an 83% survival rate and developed as well-circumscribed nodules that were predominantly composed of immature astrocytes. Solid-piece grafts in chronic contusion lesions exhibited a 92% survival rate and also developed as nodular masses. These grafts, however, contained many immature neurons 2 months postgrafting. Suspension grafts in chronic contusion lesions had an 85% survival rate and expanded in a nonrestrictive, diffuse pattern. These transplants demonstrated large neuronally rich areas of neural parenchyma. Extensive neuritic outgrowth could also be seen extending from these grafts into the surrounding host spinal cord. These findings show that human FSC tissue reliably survives and differentiates in both acute and chronic lesions. However, both the lesion environment and the grafting techniques can greatly influence the pattern of differentiation and degree of host-graft integration achieved.     

Photodynamic therapy results in induction of WAF1/CIP1/P21 leading to cell cycle arrest and apoptosis

Photodynamic therapy (PDT) is a promising new modality that utilizes a combination of a photosensitizing chemical and visible light for the management of a variety of solid malignancies. The mechanism of PDT-mediated cell killing is not well defined. We investigated the involvement of cell cycle regulatory events during silicon phthalocyanine (Pc4)-PDT-mediated apoptosis in human epidermoid carcinoma cells A431. PDT resulted in apoptosis, inhibition of cell growth, and G0-G1 phase arrest of the cell cycle, in a time-dependent fashion. Western blot analysis revealed that PDT results in an induction of the cyclin kinase inhibitor WAF1/CIP1/p21, and a down-regulation of cyclin D1 and cyclin E, and their catalytic subunits cyclin-dependent kinase (cdk) 2 and cdk6. The treatment also resulted in a decrease in kinase activities associated with all the cdks and cyclins examined. PDT also resulted in (i) an increase in the binding of cyclin D1 and cdk6 toward WAF1/CIP1/p21, and (ii) a decrease in the binding of cyclin D1 toward cdk2 and cdk6. The binding of cyclin E and cdk2 toward WAF1/CIP1/p21, and of cyclin E toward cdk2 did not change by the treatment. These data suggest that PDT-mediated induction of WAF1/CIP1/p21 results in an imposition of artificial checkpoint at G1 --> S transition thereby resulting in an arrest of cells in G0-G1 phase of the cell cycle through inhibition in the cdk2, cdk6, cyclin D1, and cyclin E. We suggest that this arrest is an irreversible process and the cells, unable to repair the damages, ultimately undergo apoptosis.     

Scaffolding: a broader view

PURPOSE: To define measures of assessing success and subsequent ways to improve excimer laser treatment of astigmatism. METHODS: We studied 97 eyes of 79 patients, followed for 12 months, that underwent photorefractive keratectomy (PRK) for myopia and astigmatism with a VISX 20/20 excimer laser. Preoperative spherical equivalent refraction at the corneal plane was between -1.00 and -15.00 D. Mean preoperative refractive astigmatism at the spectacle plan was -2.17 +/- 1.05 D (range, -1.25 to -6.00 D), which is -1.81 +/- 0.86 D (range -1.04 to -4.97 D) when calculated at the corneal plane. All patients were examined before and after surgery; examination included refraction, keratometry, and topography measurement. RESULTS: The success in treatment of astigmatism appeared measurably less than the treatment of sphere when analogous indices were used for assessment. Success in astigmatism surgery improved, as measured by all parameters, after an additional 20% was applied to astigmatism treatment magnitude indicated by the VISX computer algorithm. The sequential modes of treatment undercorrected astigmatism magnitude to a greater extent than elliptical, but equivalent success rates were present in view of the greater astigmatic changes attempted using the sequential mode. The elliptical mode tended to produce a greater undercorrection of associated sphere (p = 0.313). Results measured by refraction showed a larger change than those measured by topography and keratometry. CONCLUSION: During PRK with the VISX 20/20 laser, adjustment for undercorrection of astigmatism treatment achieves a fuller correction of astigmatism. When measuring astigmatic changes, results are different when comparing refractive astigmatism changes with corneal astigmatism changes measured by keratometry and topography.