No abstract

Cytoplasmic dynein is the only known kinetochore protein capable of driving chromosome movement toward spindle poles. In grasshopper spermatocytes, dynein immunofluorescence staining is bright at prometaphase kinetochores and dimmer at metaphase kinetochores. We have determined that these differences in staining intensity reflect differences in amounts of dynein associated with the kinetochore. Metaphase kinetochores regain bright dynein staining if they are detached from spindle microtubules by micromanipulation and kept detached for 10 min. We show that this increase in dynein staining is not caused by the retraction or unmasking of dynein upon detachment. Thus, dynein genuinely is a transient component of spermatocyte kinetochores.We further show that microtubule attachment, not tension, regulates dynein localization at kinetochores. Dynein binding is extremely sensitive to the presence of microtubules: fewer than half the normal number of kinetochore microtubules leads to the loss of most kinetochoric dynein. As a result, the bulk of the dynein leaves the kinetochore very early in mitosis, soon after the kinetochores begin to attach to microtubules. The possible functions of this dynein fraction are therefore limited to the initial attachment and movement of chromosomes and/or to a role in the mitotic checkpoint.

SummaryBackground Expression of P2X 7 , an ATP-gated calcium channel, increases cancer cell proliferation and invasiveness. A variant of P2X 7 (termed nfP2X 7 ), in which a normally hidden epitope (E200) is exposed for antibody binding, is observed in a variety of different cancers. Objectives To investigate the safety, tolerability and pharmacokinetics and assess indicative efficacy of a novel antibody ointment as a therapeutic for basal cell carcinoma (BCC). Methods An open-label, phase I clinical trial was undertaken at three dermatology clinics to evaluate the safety and tolerability of topical administration of an ointment containing 10% sheep polyclonal anti-nfP2X 7 antibodies (BIL010t) to primary BCC lesions twice daily for 28 days. Twenty-one patients with primary BCC lesions at least 0Á5 cm 2 in area and less than 2Á0 cm in diameter were enrolled. The primary end points were safety, tolerability and pharmacokinetics. Change in lesion size after treatment was determined and histology was performed on pretreatment and end-of-treatment (EOT) biopsies. Results Compliance was very high, with treatment being well tolerated. The most common adverse events were treatment site erythema, pruritus, dryness and pain. There was no evidence of systemic penetration of the sheep antibody. Lesions were measured prior to and after 28 days of treatment, with 65% of patients showing a reduction in lesion area, 20% showing no change and 15% showing an increase. Histopathology of post-treatment excision of lesion sites showed eight patients with stable disease, nine with partial response and three with complete response. Conclusions Antibodies against nfP2X 7 (BIL010t) provide a novel, safe and welltolerated treatment for BCC.

Aim To determine whether there is an association between complement factor H (CFH) or LOC387715 genotypes and response to treatment with photodynamic therapy (PDT) for exudative age-related macular degeneration (AMD). Methods Sixty-nine patients being treated for neovascular AMD with PDT were genotyped for the CFH Y402H and LOC387715 A69S polymorphisms by allele-specific digestion of PCR products. AMD phenotypes were characterized by clinical examination, fundus photography, and fluorescein angiography. Results Adjusting for age, pre-PDT visual acuity (VA), and lesion type, mean VA after PDT was significantly worse for the CFH TT genotype than for the TC or CC genotypes (P ¼ 0.05). Post-PDT VA was significantly worse for the CFH TT genotype in the subgroup of patients with predominantly classic choroidal neovascular lesions (P ¼ 0.04), but not for the patients with occult lesions (P ¼ 0.22). For the LOC387715 A69S variant, there was no significant difference among the genotypes in response to PDT therapy. Conclusions The CFH Y402H variant was associated with a response to PDT treatment in this study. Patients with the CFH TT genotype fared significantly worse with PDT than did those with the CFH TC and CC genotypes, suggesting a potential relationship between CFH genotype and response to PDT.

This study describes the common symptoms in children and adolescents following a traumatic brain injury (TBI) as reported by their primary caregivers. Utilizing data from a large-scale state demonstration project, 681 children who had sustained a TBI were ascertained from both Hospital Emergency Departments (n = 409) and Pediatric Inpatient settings (n = 272). The sample ranged in age from infancy to 18 years, was largely male (59.7%), and had equal numbers of Caucasian and minority patients. Most of the participants experienced a mild TBI (83%), with about 5.1% being moderate and 12% severe. Caregivers described the presence of current symptoms (neurological, neurocognitive, behavioral, school problems) using a series of dichotomous questions regarding their child via a structured telephone interview at 1, 4, and 10 months post-injury. Inpatient children were described as manifesting more symptoms at each of the follow-up time points than their ED counterparts. At 1 month inpatients were described as having more symptoms across all 4 domains. At 4 and 10 months, the inpatients were described as having more neurocognitive symptoms and as not returning to school on a full-time basis, with behavior problems approaching significance at the 10-month point. A large number of individuals from both groups also reported persistent symptoms 10 months post-injury including headaches, attention and memory problems, low frustration tolerance, sleep problems, personality changes, and new school problems. Practical implications of these findings for the ongoing management of these children are discussed.