Changes to insulin sensitivity in glucose clearance systems and redox following dietary supplementation with a novel cysteine-rich protein: A pilot randomized controlled trial in humans with type-2 diabetes.

We recently developed a novel keratin-derived protein (KDP) rich in cysteine, glycine, and arginine, with the potential to alter tissue redox status and insulin sensitivity. The KDP was tested in 35 human adults with type-2 diabetes mellitus (T2DM) in a 14-wk randomised controlled pilot trial comprising three 2×20 g supplemental protein/day arms: KDP-whey (KDPWHE), whey (WHEY), non-protein isocaloric control (CON), with standardised exercise. Outcomes were measured morning fasted and following insulin-stimulation (80 mU/m2/min hyperinsulinaemic-isoglycaemic clamp). With KDPWHE supplementation there was good and very-good evidence for moderate-sized increases in insulin-stimulated glucose clearance rate (GCR; 26%; 90% confidence limits, CL 2%, 49%) and skeletal-muscle microvascular blood flow (46%; 16%, 83%), respectively, and good evidence for increased insulin-stimulated sarcoplasmic GLUT4 translocation (18%; 0%, 39%) vs CON. In contrast, WHEY did not effect GCR (-2%; -25%, 21%) and attenuated HbA1c lowering (14%; 5%, 24%) vs CON. KDPWHE effects on basal glutathione in erythrocytes and skeletal muscle were unclear, but in muscle there was very-good evidence for large increases in oxidised peroxiredoxin isoform 2 (oxiPRX2) (19%; 2.2%, 35%) and good evidence for lower GPx1 concentrations (-40%; -4.3%, -63%) vs CON; insulin stimulation, however, attenuated the basal oxiPRX2 response (4%; -16%, 24%), and increased GPx1 (39%; -5%, 101%) and SOD1 (26%; -3%, 60%) protein expression. Effects of KDPWHE on oxiPRX3 and NRF2 content, phosphorylation of capillary eNOS and insulin-signalling proteins upstream of GLUT4 translocation AktSer437 and AS160Thr642 were inconclusive, but there was good evidence for increased IRSSer312 (41%; 3%, 95%), insulin-stimulated NFκB-DNA binding (46%; 3.4%, 105%), and basal PAK-1Thr423/2Thr402 phosphorylation (143%; 66%, 257%) vs WHEY. Our findings provide good evidence to suggest that dietary supplementation with a novel edible keratin protein in humans with T2DM may increase glucose clearance and modify skeletal-muscle tissue redox and insulin sensitivity within systems involving peroxiredoxins, antioxidant expression, and glucose uptake.

Rates of referable eye disease in the Scottish National Diabetic Retinopathy Screening Programme.

AIMS: Diabetic retinopathy screening aims to detect people at risk of visual loss due to proliferative diabetic retinopathy, but also refers cases of suspected macular oedema (maculopathy). At the introduction of screening, ophthalmology was concerned that referral rates would be unmanageable. We report yield of referable disease by referral reason for the first 5 years of the programme. METHODS: We extracted screening results from a nationwide clinical diabetes database to calculate annual referral rates to ophthalmic clinics. We used logistic regression to examine associations between clinical measures and referable disease. RESULTS: 182 397 people underwent ≥ 1successful retinal screening between 2006 and 2010. The yield of referable eye disease was highest in the first 2 years of screening (7.0% and 6.0%) before stabilising at ∼4.3%. The majority of referrals are due to maculopathy with 73% of referrals in 2010 based on a finding of maculopathy. CONCLUSIONS: The commonest cause for referral is for suspected macular oedema (maculopathy). Referral rates for retinopathy have stabilised, as predicted, at relatively low rates. However, ophthalmology workload continues to rise as new treatment options (ie, monthly intraocular injections) have unexpectedly increased the impact on ophthalmology. A review of the screening referral path for maculopathy may be timely.

Overreporting of vitamin D deficiency with the Roche Elecsys Vitamin D3 (25-OH) method.

BACKGROUND: Vitamin D deficiency is common. Recently Roche Diagnostics removed their Elecsys Vitamin D3 (25OH) electrochemiluminescence immunoassay (ECLIA) from use, citing deteriorating traceability to the reference method (liquid chromatography tandem mass spectrometry; LCMSMS). We investigated the performance of the Roche assay (2 assay formulations) against an LCMSMS method and the widely used DiaSorin radioimmunoassay (RIA) method. METHODS: Two sets of samples from separate populations were assayed for vitamin D. The first set was assayed using three different methods: RIA (DiaSorin) in 2004, polyclonal ECLIA (Roche) in early 2009 and LCMSMS in early 2010. The second set was assayed using polyclonal and monoclonal ECLIA (Roche) and LCMSMS in mid-2010. RESULTS: The correlation of the polyclonal ECLIA with the RIA was poor (ECLIA = 0.45 × RIA + 19, r(2) = 0.59, n = 773). LCMSMS results correlated with RIA (RIA = 0.86 × LCMSMS + 4, r(2) = 0.69, n = 49) better than with polyclonal ECLIA (polyclonal ECLIA = 0.55 × LCMSMS + 6, r(2) = 0.62, n = 55) despite a storage interval of 6 years.In recently collected samples monoclonal and polyclonal immunoassays gave similar results (monoclonal ECLIA = 0.93 polyclonal ECLIA -3, r(2) = 0.60, n = 153). The correlation between monoclonal Roche ECLIA and LCMSMS in these samples was very poor (monoclonal ECLIA = 0.31 × LCMSMS + 23, r(2) = 0.27). CONCLUSIONS: At the time of its removal from the market, the Roche Elecsys Vitamin D3 (25OH) assay showed unacceptable performance, underestimating vitamin D levels. It seems that this bias preceded the introduction of the monoclonal assay. The worldwide distribution of the assay and the duration of this bias likely led to a significant number of patients starting supplementation unnecessarily.

The mouse guanylate kinase double mutant E72Q/D103N is a functional adenylate kinase.

Guanylate kinase catalyzes the phosphorylation of either GMP to GDP or dGMP to dGDP and is an important enzyme in nucleotide metabolic pathways. Because of its essential intracellular role, guanylate kinase is a target for a number of cancer chemotherapeutic agents such as 6-thioguanine and 8-azaguanine and is involved in antiviral drug activation. Guanylate kinase shares a similarity in function and structure to other nucleoside monophosphate kinases especially with that of the well-studied adenylate kinase. Amino acid substitutions were made within the GMP binding site of mouse guanylate kinase to alter the polarity of the side chains that interact with GMP as a means of evaluating the role that these residues play on substrate interaction. One of these mutants, E72Q/D103N, was shown by functional complementation and enzyme assays to embody both guanylate kinase activity and a novel adenylate kinase activity.

The management of oral lichen planus.

Oral lichen planus is a relatively common inflammatory disease affecting between 0.5% and 2.2% of the population in epidemiological studies. In contrast with cutaneous lichen planus (LP), in which the clinical course is often mild and resolves within 2 years, mucosal LP tends to follow a more chronic course often punctuated by acute exacerbations. Furthermore, although distinct clinical subtypes such as reticular, atrophic, hypertrophic and erosive forms are well recognized, more than one clinical phenotype may be seen at a time. The rare association with oral neoplasia should always be considered and high-risk patients must be kept under close observation. Thus the management of this disorder will vary widely both between patients, and for individual patients, with fluctuations in disease activity. Here we discuss the therapeutic options available and review the evidence for their use.

Generalised granuloma annulare successfully treated with PUVA.

Disseminated granuloma annulare is an uncommon disorder in which both topical and systemic therapy may have limited success. Anecdotal reports have suggested that PUVA may result in complete clearance of disease; however, maintenance PUVA therapy has usually been required in order to maintain remission. We report the successful treatment of a patient with 5-methoxypsoralen over a 7-month period who remained in remission during a 20-month follow up period.

Biomarker assessment of the effects of petroleum refinery contamination on channel catfish.

A suite of biomarkers was used to evaluate acute (1- and 7-day) heavy metal-induced toxicity in channel catfish, Ictalurus punctatus, caged at an abandoned petroleum refinery and a non-contaminated reference site. Assays performed include indicators of metabolic, hematological, osmoregulatory, and genotoxic stress. Two cage designs were utilized to evaluate the importance of exposure routes: one allowing exclusive contact with the water column and the other allowing contact with water and sediments. Data collected at 1 and 7 days postexposure indicated that the experimental fish were stressed, having significant increases in blood glucose. After 7 days of exposure, two hemodynamic parameters, hemoglobin and delta-aminolevulinic acid dehydratase, were significantly increased. There were no significant differences observed in other parameters measured.

Symmetry and synkinesis during rehabilitation of unilateral facial paralysis.

We evaluated biofeedback rehabilitation in patients with severe chronic unilateral facial paralysis, who had intact facial-motor innervation (House grades 3 to 5). Recovery of facial function was characterized (1) by grading facial movement symmetry, and (2) by counting the number of muscles exhibiting synkinesis during maximal execution of selected facial movements (e.g., smiling). Facial function in 21 patients typically improved by one House grade. Facial symmetry recovered rapidly during the first 5 months of treatment, and then improved more slowly. However, during this latter period, examination of the relationship between symmetry and synkinesis (visualized by a graph plotting symmetry grades on the x-axis, against the number of synkinetic muscles on the y-axis) indicated that overall facial control was improving even when House grading suggested that it was not. Such information should aid facial retraining and may clarify understanding of underlying rehabilitation mechanisms.

Minimizing synkinesis during rehabilitation of the paralyzed face: preliminary assessment of a new small-movement therapy.

Neuromuscular rehabilitation can reduce the severity of chronic facial paralysis, but complete recovery is frequently impeded by synkinesis. We evaluated whether or not such synkinesis could be minimized by preventing its possible reinforcement during rehabilitation. We compared "standard" therapy, which uses the appearance of synkinesis to guide rehabilitation, with a new "small-movement" therapy, which uses smaller movements that should minimize possible subthreshold reinforcement of synkinesis. Ten subjects who had had facial paralysis for 0.5 to 27 years were randomly assigned to either therapy group. Blinded assessments were performed before and after ten 1-hour treatments given over a 1-month interval. Facial movements in both groups were significantly more symmetric after treatment. Although synkinesis tended to be reduced in the small-movement group, this reduction was not significant. The new therapy was at least as good as the standard one, and it may be better. Further studies are required to demonstrate this.

Dichotic interference effects in gap detection.

Thresholds for detecting a temporal gap in a 20-Hz-wide band of noise (the target) were measured for the target alone, and in the presence of multiple 20-Hz-wide flanking bands presented to the opposite ear. The flanking bands caused gap thresholds to increase, and this effect was greater at higher levels of the flanking bands. The impairment to gap detection was greater when the flanking bands were comodulated with the target (i.e., had the same envelope) than when they were not comodulated, except at very low and high levels of the flanking bands. A series of supplementary experiments was conducted to investigate why the difference between comodulated and noncomodulated bands was reduced at high levels. The results suggest that this was not due to inter-aural crosstalk. It may have been partly caused by: (1) a central masking effect that reduced the effective sensation level of the target band at high levels of the contralateral flanking bands; (2) reduced independence of the flanking bands owing to broadening of the auditory filters at high levels. The results are discussed in terms of perceptual grouping processes.

Zinc supplements and serum lipids in young adult white males.

A 12-wk double-blind study was conducted to determine the effect of oral zinc supplementation upon serum total cholesterol, lipoprotein-cholesterol fractions, and serum triglycerides in white males. The subjects were randomly assigned to one of three treatment groups and consumed either a placebo tablet (n = 9), 50 mg Zn/d (n = 13), or 75 mg Zn/d (n = 9) as Zn gluconate. Serum total cholesterol, low-density-lipoprotein (LDL) cholesterol, very-low-density-lipoprotein (VLDL) cholesterol, and triglycerides were not affected by Zn supplements. However, serum high-density-lipoprotein (HDL)-cholesterol levels in subjects assigned to the 75 mg Zn/d group were significantly lower at weeks 6 and 12 than those for the placebo group and lower at weeks 6, 8, and 12 than at baseline; subjects assigned to the 50 mg Zn/d group had lower serum HDL-cholesterol levels at week 12 than did the placebo group and lower at week 12 than at base line.

Intussuscepted Meckel diverticulum: radiologic-pathologic correlation.

Nine cases of Meckel diverticulum presenting with intussusception were reviewed and correlated clinically, radiologically and pathologically. The appearance of intussuscepted Meckel diverticulum by small bowel series is not specific. However, by CT, a central fat density surrounded by a thick collar of soft tissue was noted in one case correlating well with the gross pathologic appearance. The pathologic material in all our cases suggests that this CT appearance is characteristic of an inverted Meckel diverticulum.

Oligomerization and the sensitivity of phosphoenolpyruvate carboxylase to inactivation by proteinases.

Phosphenolpyruvate (PEP) carboxylase from leaves of Crassula argentea displays varying levels of sensitivity to inactivation by various proteolytic enzymes. In general, the native enzyme is sensitive to proteinases known to attack at the carbonyl end of lysine or arginine (trypsin, papain, or bromelain). The ineffective proteolytic enzymes are those which have low specificity or which attack at the N-terminal end of hydrophobic amino acids, or which cannot attack lysine. The lack of an effect of endoproteinase arginine C, which is specific for arginine, probably indicates that lysine is the critical residue. When the native enzyme, which is comprised of an equilibrium of dimers with tetramers in approximately equal quantities, is treated by preincubation with 5 millimolar PEP, the enzyme becomes much more resistant to proteolytic inactivation. When the preincubation is with 5 millimolar malate rather than buffer alone, the effect is to slightly increase (ca. 15%) the sensitivity of the enzyme to inactivation by trypsin as measured by estimates of the pseudo-first order rate constant for inactivation. PEP carboxylase from corn leaves appears to be relatively susceptible to inactivation by trypsin, but is unaffected by preincubation with malate or PEP. The sensitivity of this C(4) enzyme to inhibition by malate is also unaffected by preincubation with these ligands.

Nicotinic acid or N-methyl nicotinamide prolongs elevated brain dopa and dopamine in L-dopa treatment.

A peripheral dopa decarboxylase inhibitor, benserazide, was given ip, followed by intubation with L-dopa. Brain dopa and DA levels were elevated maximally between 0.5-2.5 hr and 1.0-2.5 hr, respectively. Dopa in serum, liver, and brain were at control values after 4 hr. Supplementation of dopa with NAM or NAC, as possible methyl group acceptors to lower catabolism of DA, showed that NAM had no effect on DA levels or on SAM. However, with both NAC and N-methyl NAM (a methylated compound intended as a control) at time periods where dopa and DA were normally decreasing, the brain levels were increased over control values with benserazide and dopa alone. NAC or N-methyl NAM appeared to extend the period of elevated brain DA levels with L-dopa treatment. The mechanism responsible for these results is uncertain.

A case-control study of caffeine and methylxanthines in benign breast disease.

A dietary case-control study of 854 histologically diagnosed cases of benign breast disease (BBD), 755 matched surgical controls, and 723 matched neighborhood controls was conducted in Israel between 1977 and 1980. No association between coffee consumption and BBD was found. Analyses by histological type, degree of ductal atypia, age, sex, and ethnic origin, controlling for several confounding factors, confirmed the lack of association. The estimated mean intake of methylxanthines was also similar for cases and controls (302, 312, and 313 mg for cases, surgical controls, and neighborhood controls, respectively). No evidence of a dose-response was noted. Our results suggest that there is no association between coffee or methylxanthine consumption and BBD, although we had a 70% chance of finding a risk ratio of 1.5 with an error of 5%.