Structure and assembly of hemidesmosomes

The hemidesmosome is a complex junction containing many proteins. The keratin cytoskeleton attaches to its cytoplasmic plaque, while its transmembrane elements interact with components of the extracellular matrix. Hemidesmosome assembly involves recruitment of alpha 6 beta 4 integrin heterodimers, as well as cytoskeletal elements and cytoskeleton-associated proteins to the cell surface. In our cell culture models, these phenomena appear to be triggered by laminin-5 in the extracellular matrix. Cell interaction with laminin-5 apparently induces both phosphorylation and dephosphorylation of subunits of alpha 6 beta 4 integrin. There is emerging evidence that such events are necessary for subsequent cytoskeleton anchorage to the hemidesmosome cytoplasmic plaque. Once assembled, the hemidesmosome plays an essential role in maintaining firm epithelial adhesion to the basement membrane, with hemidesmosome disruption being a hallmark of certain devastating blistering diseases. However, the hemidesmosome is more than just a stable anchor, as it may also be the site of signal transduction, mediated by its alpha 6 beta 4 integrin component. This review discusses our current knowledge of the structure and assembly of the hemidesmosome.     

Genomic organization of six tomato polygalacturonases and 5'' upstream sequence identity with tap1 and win2 genes

A growing number of residency programs are preparing their graduates for the realities of managed care practice. In 1996, The Cleveland Clinic Foundation, a private, nonprofit academic medical center, hosted a two-day conference on managed care education to develop innovative instructional and evaluative approaches that, where appropriate, would build on existing expertise. The conference was attended by invited national experts who had a stake in residents' education: clinical faculty, residents, medical educators, executives of managed care organizations, and representatives of other interested organizations. Participants spent much of their time in four small break out groups, each focusing on one of the following topics that were judged particularly relevant to managed care: preventive and population-based medicine, appropriate utilization of resources, clinician-patient communication, and interdisciplinary team practice. Participants shared existing materials, discussed teaching goals and objectives, and generated ideas for teaching methods, teaching materials, and evaluative methods for their respective topics. The authors summarize the recommendations from the four groups, with an overview of the issues that emerged during the conference concerning curriculum development, integration of managed care topics into existing curricula, staging of the curriculum, experiential teaching methods, negative attitudes and resistance, evaluation of trainees and profiling, program assessment, faculty development, and cooperation between academic medical centers and managed care organizations.     

The Ile164 beta2-adrenergic receptor polymorphism adversely affects the outcome of congestive heart failure

The beta2-adrenergic receptor (beta2AR), an important modulator of cardiac inotropy and chronotropy, has significant genetic heterogeneity in the population. Because dysfunctional betaARs play a role in the pathogenesis of the failing ventricle, we tested the hypothesis that beta2AR polymorphisms alter the outcome of congestive heart failure. 259 patients with NYHA functional class II-IV heart failure due to ischemic or dilated cardiomyopathy were genotyped and prospectively followed, with the endpoint defined as death or cardiac transplantation. The allele frequencies between this group and those of 212 healthy controls also were compared and did not differ between the groups. However, those with the Ile164 polymorphism displayed a striking difference in survival with a relative risk of death or cardiac transplant of 4.81 (P < 0.001) compared with those with the wild-type Thr at this position. Age, race, gender, functional class, etiology, ejection fraction, and medication use did not differ between these individuals and those with the wild-type beta2AR, and thus the beta2AR genotype at position 164 was the only clear distinguishing feature between the two groups. The 1-yr survival for Ile164 patients was 42% compared with 76% for patients harboring wild-type beta2AR. In contrast, polymorphisms at amino acid positions 16 (Arg or Gly) or 27 (Gln or Glu), which also alter receptor phenotype, did not appear to have an influence on the course of heart failure. Taken together with cell-based and transgenic mouse results, this study establishes a paradigm whereby genetic variants of key signaling elements can have pathophysiologic consequences within the context of a disease. Furthermore, patients with the Ile164 polymorphism and heart failure may be candidates for earlier aggressive intervention or cardiac transplantation.     

Decrease of capsular opacification with adjunctive mitomycin C in combined glaucoma and cataract surgery

OBJECTIVE: The authors investigated the incidence of capsular opacification requiring YAG capsulotomy after primary trabeculectomy combined with phacoemulsification and implantation of all polymethylmethacrylate intraocular lenses. DESIGN: A prospective randomized study. PARTICIPANTS: One hundred seventy-four eyes of 174 nonselected patients with primary open-angle glaucoma (POAG) were randomized to either no adjunctive mitomycin C (MMC) control group of 93 eyes of 93 patients) or adjunctive subconjunctival MMC (MMC group of 81 eyes of 81 patients) during the primary glaucoma triple procedure (PGTP). INTERVENTION: Primary glaucoma triple procedure with and without MMC and YAG laser capsulotomy for posterior capsular opacification (PCO) was performed. MAIN OUTCOME MEASURES: The incidences of YAG capsulotomy for PCO were compared between the control and MMC groups and also between the control group and the MMC subgroups (1 minute, 3 minutes, and 5 minutes of MMC application) using Kaplan-Meier analysis with Mantel-Cox log-rank test. Cox proportional hazard regression analysis also was performed to identify significant factors affecting capsular opacification. RESULTS: The control and MMC groups were similar in preoperative characteristics. However, the probability of PCO requiring YAG capsulotomy was significantly lower in the MMC group than in the control group (P = 0.004). Among the MMC subgroups, MMC application for 3 minutes was most effective and significant when compared with that of the control group (P = 0.002). Although not as significant as the intraoperative use of MMC (P = 0.002), old age (P = 0.026) and presence of diabetes mellitus (P = 0.035) were also identified as significant beneficial factors for decreasing the incidence of YAG capsulotomy for PCO in Cox proportional hazard regression analysis. CONCLUSION: Intraoperative subconjunctival MMC application during combined glaucoma and cataract surgery has a beneficial effect of inhibiting PCO after combined surgery in patients with POAG. Thus, after intraoperative subconjunctival application of MMC at the concentration of 0.5 mg/ml for 3 minutes, the aqueous MMC level must have been great enough to inhibit the lens epithelial cell proliferation to result in a long-term decrease in PCO.     

Interaction of Arabidopsis kinesin-like calmodulin-binding protein with tubulin subunits: modulation by Ca(2+)-calmodulin

Kinesin-like calmodulin-binding protein (KCBP) is a recently identified novel kinesin-like protein that appears to be unique to and ubiquitous in plants. KCBP is distinct from all other known KLPs in having a calmodulin-binding domain adjacent to its motor domain. We have used different regions of KCBP to study its interaction with tubulin subunits and the regulation of this interaction by Ca(2+)-calmodulin. The results show that the carboxy-terminal part of the KCBP, with or without calmodulin-binding domain, binds to tubulin subunits and this binding is sensitive to nucleotides. In the presence of Ca(2+)-calmodulin the motor with calmodulin-binding domain does not bind to tubulin. This Ca(2+)-calmodulin modulation is abolished in the presence of antibodies specific to the calmodulin-binding domain of KCBP. Similar binding studies with the carboxy-terminal part of KCBP lacking the calmodulin-binding domain show no effect of Ca(2+)-calmodulin. These results indicate that Ca(2+)-calmodulin modulates the interaction of KCBP with tubulin subunits and this modulation is due to the calmodulin-binding domain in the KCBP. Calcium-dependent calmodulin modulation of KCBP interaction with tubulin suggests regulation of KCBP function by calcium, the first such regulation of a kinesin heavy chain among all the known kinesin-like proteins.     

Characterization of microtubule binding domains in the Arabidopsis kinesin-like calmodulin binding protein

The kinesin-like calmodulin binding protein (KCBP) is a new member of the kinesin superfamily that appears to be present only in plants. The KCBP is unique in its ability to interact with calmodulin in a Ca2+-dependent manner. To study the interaction of the KCBP with microtubules, we expressed different regions of the Arabidopsis KCBP and used the purified proteins in cosedimentation assays with microtubules. The motor domain with or without the calmodulin binding domain bound to microtubules. The binding of the motor domain containing the calmodulin binding region to microtubules was inhibited by Ca2+-calmodulin. This Ca2+-calmodulin regulation of motor domain interactions with microtubules was abolished in the presence of antibodies specific to the calmodulin binding region. In addition, the binding of the motor domain lacking the calmodulin binding region to microtubules was not inhibited in the presence of Ca2+-calmodulin, suggesting an essential role for the calmodulin binding region in Ca2+-calmodulin modulation. Results of the cosedimentation assays with the N-terminal tail suggest the presence of a second microtubule binding site on the KCBP. However, the interaction of the N-terminal tail region of the KCBP with microtubules was insensitive to ATP. These data on the interaction of the KCBP with microtubules provide new insights into the functioning of the KCBP in plants.     

The role of Staphylococcus epidermidis persistence factors in the infectious process

The species composition of microflora was determined and the etiological role of S. epidermidis was shown in some purulent inflammatory diseases (maxillary sinusitis, urethritis). The role of the complex of S. epidermidis persistence factors in the chronization of the infectious process and the formation of the resident carrier state was established.