Neuropeptide Y: some viewpoints on a multifaceted peptide in the normal and diseased nervous system

Using immunohistochemical and in situ hybridization methodologies the localization of neuropeptide tyrosine (NPY) and two of its receptors, the Y1- and the Y2-receptor (R), has been analysed in various tissues in normal animals and animals subjected to different experimental procedures as well as animals with a genetic and an acquired disease. (1) Dorsal root ganglion (DRG) neurons are discussed with special focus on the effect of peripheral nerve injury. In normal DRG neurons NPY cannot be detected, whereas Y1-R mRNA and Y1-R-like immunoreactivity (LI) are strongly expressed. The Y1-Rs decorate the membrane of the cell soma and are not transported peripherally into the axonal branches. Y2-R mRNA levels are low. After axotomy there is a marked increase in NPY, a decrease in Y1-Rs and an increase in Y2-Rs. The Y2-R is transported centrifugally. These findings suggest that NPY-ergic mechanisms participate in the adaptive changes of sensory neurons in response to injury. (2) Using specific antibodies the cellular and subcellular localization of the Y1-R protein have been analysed in cerebral blood vessels. The results demonstrate high concentrations of receptors in smooth muscle cells around pial arterioles with lower numbers in large vessels on the basal surface of the brain. In many regions the receptors 'disappear' after the arterioles have entered the brain tissue. At the ultrastructural level the receptors are found both on the endothelial and peripheral side of the muscle cells as well as laterally, where muscle cells oppose each other. The receptor protein is often associated with small vesicles. No NPY-positive nerve fibers were found around the Y1-R-rich arterioles, but they were only seen around the arteries with low Y1-R levels. The Y1-R-rich arterioles were, however, seen close to numerous NPY-positive fibers originating from central interneurons. These findings raise the possibility that centrally originating NPY can influence cerebral blood flow, possibly by stimulating NPY-Rs on the peripheral side of the muscle cells. However, also blood borne NPY, released under special conditions, such as stress from sympathetic nerves and the adrenal medulla and transported with blood, may stimulate receptors on the endothelial side of the smooth muscle cells. (3) In the arcuate nucleus Y1- and Y2-Rs are found, whereby the Y1-Rs are located in its ventro-medial portion and co-localized with POMC peptides, and the Y2-R in its ventromedial part, partly co-localized with NPY. NPY nerve endings makes synaptic contact with the POMC/Y1-R-positive neurons. In a mouse model for genetic anorexia very high levels of NPY were observed in arcuate neurons as compared to control mice. However, NPY mRNA levels were not different between the two groups. Taken together these findings are in good agreement with the view that NPY in the arcuate nucleus plays an important role in regulating feeding behaviour. (4) After intracerebral prion inoculation in mice an upregulation of NPY mRNA levels was observed in CA3 pyramidal neurons, and this effect was seen at a time point just before the first behavioural symptoms were manifested. At approximately the same time there was a dramatic decrease in Y2-R binding in strata oriens and radiatum of the CA1 region of the hippocampus, whereas in other regions no changes or much smaller changes were observed. Also, there was only a very slight decrease in Y2-R mRNA levels in CA3 neurons. It thus appears as if the prion disease prevents ligand binding to the Y2-R, perhaps by influencing traffic of receptor proteins, possibly at the level of cell membrane-associated caveolae, which have been implicated in the conversion of normal protein to scrapie protein. It is possible that these changes in NPY-ergic mechanisms may underlie some of the central symptoms associated with the prion disease. (ABSTRACT TRUNCATED)     

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.     

Predictors of early failure of fixation in the treatment of displaced subcapital hip fractures

Malignant hyperthermia is a rare complication in clinical anesthesia, especially associated with the administration of succinylcholine or inhalation anesthetics. A 19-year-old patient, suffering from traumatic mandible fracture, underwent open reduction under general anesthesia. Unfortunately, following administration of succinylcholine, he also suffered severe facial twitch and the first episode of hypercapnia. After adequate management, the symptoms subsided. However, two hours later, the hypercapnia recurred, combined with progressive elevation of body temperature. After administration of intravenous dantrolene 120 mg, the patient's condition became stable and the procedure was completed without sequelae. As the muscle contracture test is not available in Taiwan, the clinical grading scale is presented as an alternative diagnostic method for malignant hyperthermia.