Localization and expression of clarin-1, the Clrn1 gene product, in auditory hair cells and photoreceptors

The Usher syndrome 3A (CLRN1) gene encodes clarin-1, which is a member of the tetraspanin family of transmembrane proteins. Although identified more than 6 years ago, little is known about its localization or function in the eye and ear. We developed a polyclonal antibody that react with all clarin-1 isoforms and used it to characterize protein expression in cochlea and retina. In the cochlea, we observe clarin-1expression in the stereocilia of P0 mice, and in synaptic terminals present at the base of the auditory hair cells from E18 to P6. In the retina, clarin-1 localizes to the connecting cilia, inner segment of photoreceptors and to the ribbon synapses. RT-PCR from P0 cochlea and P28 retina show mRNAs encoding only isoforms 2 and 3. Western blots show that only isoform 2 is present in protein extracts from these same tissues. We examined clarin-1 expression in the immortomouse-derived hair cell line UB/OC-1. Only isoform 2 is expressed in UB/OC-1 at both mRNA and protein levels, suggesting this isoform is biologically relevant to hair cell function. The protein co-localizes with microtubules and post-transgolgi vesicles. The subcellular localization of clarin-1 in hair cells and photoreceptors suggests it functions at both the basal and apical poles of neurosensoriepithelia.     

Elevated liver enzymes in individuals with undiagnosed diabetes in the U.S.

ObjectiveWe investigated the association of diabetes diagnosis and medication type with liver injury in individuals with clinical diabetes.MethodsWe analyzed 2426 patients with clinical diabetes in the National Health and Nutrition Examination Survey conducted from 1999 to 2008. Elevated alanine transaminase (ALT) and aspartate transaminase (AST) levels were used as markers of liver injury. Participants were categorized into one of three categories: 1) clinical diabetes without physician’s diagnosis or diabetes medication; 2) diagnosed diabetes without diabetes medication; or 3) diagnosed diabetes with diabetes medication, further divided by classes of diabetes medications prescribed. We conducted logistic regression analysis to examine the relationship between diabetes diagnosis and medication type and elevated ALT and AST, adjusting for race/ethnicity, education, health insurance status, BMI category, alcohol consumption, physical activity, antihyperlipidemic agents, glycohemoglobin, C-reactive protein, viral hepatitis and liver disease.ResultsParticipants with undiagnosed diabetes were more likely to have elevated ALT and AST levels (OR = 1.82, 95% CI 1.47, 2.42; OR = 1.99, 95% CI 1.46, 2.71, respectively). In contrast, there was no association between specific diabetes medication (i.e., sulfonylureas, biguanides/thiazolidinediones) and elevated ALT or AST levels among the treated. Our findings were confirmed in sensitivity analyses employing a lower threshold for ALT, and excluding individuals with viral hepatitis or liver disease.ConclusionWe found that undiagnosed diabetes is associated with liver injury, compared to diagnosed diabetes with treatment. The effect of diabetes treatment on liver injury in individuals with diabetes remains uncertain.     

Metabolism of oestrone and oestradiol-17beta to conjugated steroids by the accessory sex glands of the male pig.

Oestrogens are secreted in large amounts by boar testes and are known to have a synergistic effect with testosterone on the production of large volumes of seminal plasma. Thus, oestrogens play a role in regulating the large accessory sex glands in the boar. Since testosterone metabolites (e.g. 5alpha-dihydrotestosterone) account for much of its action in target tissues we have looked at the metabolism of oestrogens in the accessory sex glands of the male pig. Tissues from seminal vesicles and bulbourethral glands of 6-week-old castrate and intact males, and 12-week-old castrate animals, were incubated with (3)H-labelled oestrone and oestradiol-17beta. Aliquots of spent culture medium and of methanolic tissue extracts were taken to measure radioactivity, prior to separation of unconjugated and conjugated steroids on Waters C(18) Sep-Pak cartridges. About one-third of the radioactivity appeared as conjugates in the media from both glands with each oestrogen. Subsequently, sulphoconjugated steroids and glucuronidates were recovered in series from C(18) cartridges after solvolysis and enzyme hydrolysis respectively. Furthermore, about one-third of the conjugated fraction in each case remained unhydrolysed after these treatments. In conclusion, it is clear that a study of the actions of oestrogens on these glands must consider the dynamics of metabolism of the oestrogens presented to them by the testes and would include conjugation of steroids by the glands themselves.     

Pain, quality of life, and coping in sickle cell disease

This study examined the frequency and severity of sickle related pain, its impact on quality of life, and methods of coping for 25 children with sickle cell disease, aged 6-16 years. Subjects were matched with non-affected peers and asked to complete the Central Middlesex Hospital Children's Health Diary for four weeks. Results indicated that sickle pain occurred on average one in 14 days, and total summary pain scores indicated significantly greater pain than for controls. Children with sickle cell disease could discriminate sickle pain and did not adopt sick role responses to ordinary childhood ailments. Nearly all sickle pain was dealt with at home. Sickle pain resulted in over seven times increased risk of not attending school and was highly disruptive of social and recreational activities. Careful assessment of sickle pain in the home environment is an essential part of a community focused pain management service, which effectively supports children's resilience and improves their quality of life.