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Objectives: Phase 3 licensing trials for the recently approved respiratory syncytial virus (RSV) vaccines did not include many residents of long-term care facilities (LTCF). Our primary objective was to assess humoral immune responses in LTCF residents, age 60 and older, to the RSV vaccines, and demonstrate noninferiority to antibody responses in community-dwelling (CD) adults who were representative of the phase 3 trial participants in whom the vaccines were highly efficacious., Design: Prospective non-randomized intervention trial of RSV vaccines in LTCF residents., Setting and Participants: Research clinic and 2 LTCFs. Adults ≥60 years old, free of immunosuppression and planning to receive an RSV vaccine were eligible., Methods: LTCF and CD participants received either the GSK or Pfizer RSV vaccine in equal numbers. Blood was collected before and 30 days after vaccination. Total immunoglobulin (Ig)G to the prefusion F protein of RSV group A (F A ) and B (F B ), and neutralizing activity were measured, and geometric mean titer (GMT) and geometric mean fold rise (GMFR) calculated. Intercurrent respiratory illnesses were tracked., Results: A total of 76 LTCF residents and 76 CD adults were enrolled. Day 30 blood was unavailable from 3 residents and 3 had RSV infection and vaccination was deferred, leaving data for 76 CD and 70 LTCF adults for analysis. Serum IgG GMFR prefusion F A (9.9 vs 12.5, P = .14), prefusion F B (8.7 vs 11.0, P = .17) were not statistically different in CD and LTCF cohorts, respectively, and also equivalent for GMFR in viral neutralization titers (12.8 vs. 15.5, P = .32). As measured by GMT or GMFR, RSV vaccine responses of LTCF residents met noninferiority criteria compared with the CD cohort., Conclusions and Implications: This small immunobridging study demonstrates robust antibody responses to RSV vaccines in LTCF residents providing support for their use in this high-risk population., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Americans consume diets that fall short of dietary recommendations, and the cost of healthier diets is often cited as a barrier to dietary change. We conducted a nonrandomized crossover trial with meals provided utilizing 2 diets: Dietary Approaches to Stop Hypertension (DASH) and whole food, plant-based (WFPB), and thus had intake data from baseline and both intervention diets., Objectives: Using actual diet records, describe food costs of baseline diets of individuals with type 2 diabetes (T2DM) as well as therapeutic DASH and WFPB diets., Methods: Three-day food records were collected and analyzed for each 7-d diet phase: baseline, DASH, and WFPB. Nutrient content was analyzed using the Nutrient Data System for Research and cost was determined using Fillet, an application to manage menu pricing. Food costs were calculated for each diet as consumed and adjusted to a standardized 1800 kcal/d. Ingredient-only costs of food away from home (FAFH) were approximated and analyzed. Costs were analyzed using linear mixed-effect models as a function of diet., Results: Fifteen subjects enrolled; 12 completed all dietary phases. The baseline, DASH, and WFPB diets, as consumed, cost $15.72/d (95% CI; $13.91, $17.53), $12.74/d ($11.23, $14.25), and $9.78/d ($7.97, $11.59), respectively. When adjusted to an 1800 kcal/d intake, the baseline, DASH, and WFPB diets cost $15.69/d ($13.87, $17.52), $14.92/d ($13.59, $16.26), and $11.96/d ($10.14, $13.78), respectively. When approximated ingredient-only costs of FAFH were analyzed, as consumed baseline [$11.01 ($9.53, $12.49)] and DASH diets [$11.81 ($10.44, $13.18)] had similar costs; WFPB diet [$8.83 ($7.35, $10.31)] cost the least., Conclusions: In this short-term study with meals provided, the food costs of plant-predominant diets offering substantial metabolic health benefits were less than or similar to baseline food costs of adults with insulin-treated T2DM. Longer-term data without meal provision are needed for more generalizable results. This trial was registered at clinicaltrials.gov as NCT04048642., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Women with congenital long QT syndrome (LQTS) experience increased cardiac event risk after the onset of adolescence, perhaps stemming from the known modulating effects of sex hormones on the cardiac potassium channels., Objective: We hypothesized that the effect of sex hormones on cardiac ion channel function may modify electrocardiographic (ECG) parameters associated with the propensity for ventricular tachyarrhythmias during the menstrual cycle in women with LQTS., Methods: We prospectively enrolled 65 women with congenital LQTS (type 1 LQTS [LQT1], n = 24 [36.9%]; type 2 LQTS [LQT2], n = 20 [30.8%]) and unaffected female relatives (n = 21 [32.3%]). Patients underwent three 7-day ECG recordings during their menstrual cycles. Simultaneous saliva testing of sex hormone levels was conducted on the first day of each 7-day ECG recording cycle., Results: The mean age was 35 ± 8 years, without a significant difference among the groups. In women with LQT2, linear mixed effects models showed significant inverse correlations of the corrected QT interval with progesterone levels (P < .001) and with the progesterone to estradiol ratio (P < .001). Inverse relationships of the R-R interval with estradiol levels (P = .003) and of the T-wave duration with testosterone levels (P = .014) were also observed in women with LQT2. In contrast, no significant associations were observed between ECG parameters and sex hormone levels in women with LQT1 or unaffected relatives., Conclusion: This is the first study to prospectively assess correlations between repolarization dynamics and sex hormone levels during the menstrual cycle in women with congenital LQTS. Our findings show genotype-specific unique corrected QT dynamics during the menstrual cycle that may affect the propensity for ventricular tachyarrhythmia in women with LQTS, particularly women with LQT2., (Copyright © 2022. Published by Elsevier Inc.)

Marine medaka Oryzias melastigma at 4months (young), 8months (middle-aged) and 12months old (senior) were employed to determine age-associated change of sex ratios, sex hormones, telomere length (TL), telomerase activity (TA), telomerase transcription (omTERT) and oxidative damage in the liver. Overall, O. melastigma exhibited gradual senescence, sex differences in longevity (F>M), TL (F>M) and oxidative damage (F5kb), TA and omTERT expression (p≤0.01), and negatively correlated with liver DNA oxidation (p≤0.05). The results suggest high levels of E2 in female O. melastigma may retard TL shortening by enhancing TA via TERT transcription and/or reducing oxidative DNA damage. The findings support TL shortening as a biomarker of aging and further development of accelerated TL shortening, abnormal suppression of TA and excessive oxidative DNA damage as early molecular endpoints, indicative of advanced/premature aging in marine medaka/fish., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: It is estimated that 33.5 million people in the world have developed atrial fibrillation (AF), and an estimated 30% of patients with AF are unaware of their diagnosis (silent AF)., Objective: The purpose of this study was to test a new technology for contactless detection of AF based on facial video recordings., Methods: The proposed technique uses a camera to record an individual's face and extract the subtle beat-to-beat variations of skin color reflecting the cardiac pulsatile signal. In a group of adults referred for electrical cardioversion, we recorded the ECG and the video of the subjects' face before and after electrical cardioversion. We extracted the beat-to-beat pulse rates expressed as pulses per minute (ppm) from the videoplethysmographic (VPG) signal acquired using a standard web camera. We introduce a novel quantifier of pulse variability called the pulse harmonic strength (PHS) and report its ability to detect the presence of AF., Results: Eleven subjects (8 male; age 65 ± 6 years) were included in the study. The VPG and ECG-based rates were statistically different between the AF and sinus rhythm periods: 72 ± 9 ppm vs 57 ± 7 ppm (P < .0001) for VPG and 80 ± 17 bpm vs 56 ± 7 bpm (P < .0001) for ECG signals. Among the 407 epochs of 15 seconds of synchronized ECG and VPG signals, PHS was associated with a 20% detection error rate, and the error rates of the automatic ECG-based measurements ranged between 17% and 29%., Conclusion: Our preliminary results support the concept that contactless video-based monitoring of the human face for detection of abnormal pulse variability due to AF is feasible., (Copyright © 2015 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Vibration-reducing (VR) gloves have been increasingly used to help reduce vibration exposure, but it remains unclear how effective these gloves are. The purpose of this study was to estimate tool-specific performances of VR gloves for reducing the vibrations transmitted to the palm of the hand in three orthogonal directions (3-D) in an attempt to assess glove effectiveness and aid in the appropriate selection of these gloves. Four typical VR gloves were considered in this study, two of which can be classified as anti-vibration (AV) gloves according to the current AV glove test standard. The average transmissibility spectrum of each glove in each direction was synthesized based on spectra measured in this study and other spectra collected from reported studies. More than seventy vibration spectra of various tools or machines were considered in the estimations, which were also measured in this study or collected from reported studies. The glove performance assessments were based on the percent reduction of frequency-weighted acceleration as is required in the current standard for assessing the risk of vibration exposures. The estimated tool-specific vibration reductions of the gloves indicate that the VR gloves could slightly reduce (<5%) or marginally amplify (<10%) the vibrations generated from low-frequency (<25 Hz) tools or those vibrating primarily along the axis of the tool handle. With other tools, the VR gloves could reduce palm-transmitted vibrations in the range of 5%-58%, primarily depending on the specific tool and its vibration spectra in the three directions. The two AV gloves were not more effective than the other gloves with some of the tools considered in this study. The implications of the results are discussed., Relevance to Industry: Hand-transmitted vibration exposure may cause hand-arm vibration syndrome. Vibration-reducing gloves are considered as an alternative approach to reduce the vibration exposure. This study provides useful information on the effectiveness of the gloves when used with many tools for reducing the vibration transmitted to the palm in three directions. The results can aid in the appropriate selection and use of these gloves.

Fibrosis is at the core of the high morbidity and mortality rates associated with the complications of diabetes and obesity, including diabetic nephropathy (DN), without any US Food and Drug Administration-approved drugs with this specific target. We recently provided the first evidence that the matricellular protein CCN3 (official symbol NOV) functions in a reciprocal manner, acting on the profibrotic family member CCN2 to inhibit fibrosis in a mesangial cell model of DN. Herein, we used the BT/BR ob/ob mouse as a best model of human obesity and DN progression to determine whether recombinant human CCN3 could be used therapeutically, and the mechanisms involved. Eight weeks of thrice-weekly i.p. injections (0.604 and 6.04 μg/kg of recombinant human CCN3) beginning in early-stage DN completely blocked and/or reversed the up-regulation of mRNA expression of kidney cortex fibrosis genes (CCN2, Col1a2, TGF-β1, and PAI-1) seen in placebo-treated diabetic mice. The treatment completely blocked glomerular fibrosis, as determined by altered mesangial expansion and deposition of laminin. Furthermore, it protected against, or reversed, podocyte loss and kidney function reduction (rise in plasma creatinine concentration); albuminuria was also greatly reduced. This study demonstrates the potential efficacy of recombinant human CCN3 treatment in DN and points to mechanisms operating at multiple levels or pathways, upstream (eg, protecting against cell injury) and downstream (eg, regulating CCN2 activity and extracellular matrix metabolism).

Lymphoma cells are subject to higher levels of oxidative stress compared with their normal counterparts and may be vulnerable to manipulations of the cellular redox balance. We therefore designed a phase 2 study of imexon (Amplimexon/NSC-714597), a prooxidant molecule, in patients with relapsed/refractory B-cell non-Hodgkin lymphoma (NHL). Imexon was administered at 1000 mg/m(2) IV daily for 5 days in 21-day cycles. Gene expression analysis performed on pretreatment tumor specimens included 13 transcripts used to generate a redox signature score, previously demonstrated to correlate with lymphoma prognosis. Twenty-two patients were enrolled having follicular (n = 9), diffuse large B-cell (DLBCL) (n = 5), mantle cell (n = 3), transformed follicular (n = 2), small lymphocytic (n = 2), and Burkitt (n = 1) lymphoma. The most common grade 3/4 adverse events were anemia (14%) and neutropenia (9%). The overall response rate was 30%, including responses in follicular lymphoma (4 of 9) and DLBCL (2 of 5). Gene expression analyses revealed CD68 and the redox-related genes, GPX1 and SOD2, as well as a higher redox score to correlate with clinical responses. Therefore, pretreatment markers of oxidative stress may identify patients likely to respond to this therapeutic approach. This trial was registered at www.clinicaltrials.gov as #NCT01314014., (© 2014 by The American Society of Hematology.)

Background: In prior clinical studies of patients with long QT syndrome (LQTS), pregnancy was associated with fewer cardiac events (CEs) compared to before or after pregnancy. In recent animal studies involving rabbits with LQTS mutations, progesterone had favorable effects on CEs compared to estrogen. The effect of oral contraceptive therapy with its high progesterone/estrogen ratio on the risk of CEs in patients with LQTS has not been examined., Objective: To study the effect of oral contraceptive use on the risk of CEs in patients with LQTS., Methods: We studied 174 patients from the Rochester-based LQTS Registry who responded to a questionnaire about their oral contraceptive use. We used time-dependent Cox regression to estimate the hazard ratio for recurrent CEs when patients were using vs not using oral contraceptives during nonpregnancy periods. For this recurrent events analysis, the Prentice-Williams-Peterson model was used and the time origin was defined as the onset of menarche. We adjusted for the baseline corrected QT interval, history of CEs before menarche, age at menarche onset, number of births, time-dependent β-blocker therapy, and LQTS genotype., Results: No differences in the risk of CEs for the times patients with LQTS were using vs not using oral contraceptives was found in the general population with LQTS (hazard ratio 1.01; P = .95) or in analyses of LQTS subsets (P > .2)., Conclusion: Oral contraceptive therapy use did not affect LQTS-related CEs in the study population. Oral contraceptives did not show beneficial or harmful effects in this patient group., (Copyright © 2014 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Background: A prolonged QT interval corrected for heart rate (QTc) is a major risk factor in patients with long QT syndrome (LQTS). However, heart rate-related risk in this genetic disorder differs among genotypes., Objective: This study hypothesized that risk assessment in LQTS patients should incorporate genotype-specific QT correction for heart rate., Methods: The independent contribution of 4 repolarization measures (the absolute QT interval, and Bazett's, Fridericia's, and Framingham's correction formulas) to the risk of aborted cardiac arrest or sudden cardiac death during adolescence, before and after further adjustment for the RR interval, was assessed in 727 LQTS type 1 and 582 LQTS type 2 patients. Improved QT/RR correction was calculated using a Cox model, dividing the coefficient on log(RR) by that on log(QT)., Results: Multivariate analysis demonstrated that in LQTS type 1 patients 100-ms increments in the absolute QT interval were associated with a 3.3-fold increase in the risk of life-threatening cardiac events (P = .020), and 100-ms decrements in the RR interval were associated with a further 1.9-fold increase in the risk (P = .007), whereas in LQTS type 2 patients, resting heart rate was not a significant risk factor (hazard ratio 1.11; P = .51; P value for heart rate × genotype interaction = .036). Accordingly, analysis of an improved QT correction formula showed that patients with the LQTS type 1 genotype required a greater degree of QT correction for heart rate (improved QTc = QT/RR⁰·⁸) than LQTS type 2 patients (improved QTc = QT/RR⁰·²)., Conclusion: Our findings suggest that risk stratification for life-threatening cardiac events in LQTS patients can be improved by incorporating genotype-specific QT correction for heart rate., (Copyright © 2011 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Background: There is a consensus on the limited value of the QTc interval prolongation as a surrogate marker of drug cardiotoxicity and as a risk stratifier in inherited long QT syndrome (LQTS) patients., Objective: We investigated the interest of repolarization morphology in the acquired and the inherited LQTS., Methods: We analyzed 2 retrospective electrocardiographic (ECG) datasets from healthy on/off moxifloxacin and from genotyped KCNH2 patients. We measured QT, RR, and T-peak to T-end intervals, early repolarization duration (ERD) and late repolarization duration, T-roundness, T-amplitude, left (αL) and right slopes of T-waves. We designed multivariate logistic models to predict the presence of the KCNH2 mutation or moxifloxacin while adjusting for the level of QTc prolongation and the level of heart rate in LQT2 patients. Independent learning and validation sets were used. A list of 4,874 ECGs from 411 healthy individuals, 293 from 143 LQT2 carriers and 150 noncarrier family members were analyzed., Results: In the moxifloxacin model, ERD was associated with the presence of the drug (odds ratio = 1.15 per ms increase, confidence interval 1.04 to 1.26, P = .0001) after adjustment for QTc. The model for the LQT2 revealed that left slope was associated with the presence of the KCNH2 mutation (odds ratio = 0.38 per 1.5 μV/ms decrease, confidence interval 0.23 to 0.64, P = .0002). Only T-roundness complemented QTc in the model investigating cardiac events in LQT2., Conclusions: These observations demonstrate that the phenotypic expression of KCNH2 mutations and the effect of IKr-inhibitory drug on the surface electrocardiogram are specific. Future research should investigate whether this phenomenon is linked to different level/form of loss functions of Ikr channels, and whether they could result in different arrhythmogenic mechanisms., (Copyright © 2011 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

Fibrosis is a major cause of end-stage renal disease, and although initiation factors have been elucidated, uncertainty concerning the downstream pathways has hampered the development of anti-fibrotic therapies. CCN2 (CTGF) functions downstream of transforming growth factor (TGF)-beta, driving increased extracellular matrix (ECM) accumulation and fibrosis. We examined the possibility that CCN3 (NOV), another CCN family member with reported biological activities that differ from CCN2, might act as an endogenous negative regulator of ECM and fibrosis. We show that cultured rat mesangial cells express CCN3 mRNA and protein, and that TGF-beta treatment reduced CCN3 expression levels while increasing CCN2 and collagen type I activities. Conversely, either the addition of CCN3 or CCN3 overexpression produced a marked down-regulation of CCN2 followed by virtual blockade of both collagen type I transcription and its accumulation. This finding occurred in both growth-arrested and CCN3-transfected cells under normal growth conditions after TGF-beta treatment. These effects were not attributable to altered cellular proliferation as determined by cell cycle analysis, nor were they attributable to interference of Smad signaling as shown by analysis of phosphorylated Smad3 levels. In conclusion, both CCN2 and CCN3 appear to act in a yin/yang manner to regulate ECM metabolism. CCN3, acting downstream of TGF-beta to block CCN2 and the up-regulation of ECM, may therefore serve to naturally limit fibrosis in vivo and provide opportunities for novel, endogenous-based therapeutic treatments.

Recent studies have described malignant stem cells as central to the initiation, growth, and potential relapse of acute and chronic myelogenous leukemia (AML and CML). Because of their important role in pathogenesis, rare and biologically distinct leukemia stem cells (LSCs) represent a critical target for therapeutic intervention. However, to date, very few agents have been shown to directly target the LSC population. The present studies demonstrate that parthenolide (PTL), a naturally occurring small molecule, induces robust apoptosis in primary human AML cells and blast crisis CML (bcCML) cells while sparing normal hematopoietic cells. Furthermore, analysis of progenitor cells using in vitro colony assays, as well as stem cells using the nonobese diabetic/severe combined immunodeficient (NOD/SCID) xenograft model, show that PTL also preferentially targets AML progenitor and stem cell populations. Notably, in comparison to the standard chemotherapy drug cytosine arabinoside (Ara-C), PTL is much more specific to leukemia cells. The molecular mechanism of PTL-mediated apoptosis is strongly associated with inhibition of nuclear factor kappa B (NF-kappaB), proapoptotic activation of p53, and increased reactive oxygen species (ROS). On the basis of these findings, we propose that the activity of PTL triggers LSC-specific apoptosis and as such represents a potentially important new class of drugs for LSC-targeted therapy.

Renal tubular reabsorption and hydrolysis of plasma high density lipoprotein (HDL3) were studied. Rabbit proximal straight nephron segments were microperfused in vitro with iodinated HDL3. Progressive luminal uptake and cellular accumulation of radiolabeled material were observed during an initial phase, followed by a reduction in sequestration and the appearance of 125I-label in the bathing medium. To detect proteolysis, collected perfusates and bathing media were analyzed for trichloracetic acid soluble radioactivity. 125I-HDL3 in the luminal fluid was intact, but metabolites appeared in the bathing medium. Electron microscopic radioautography demonstrated endocytic uptake of 125I-HDL3 at the luminal membrane of the proximal tubule and movement of grains into lysosome-like dense bodies. Incubation of radiolabeled HDL3 in the presence of renal homogenates resulted in proteolytic activity with an acidic pH optimum. Analytical cell fractionation studies indicated that hydrolysis of the protein component is associated with lysosomes derived from proximal kidney tubules. Collectively, the data show that plasma HDL3 can be reabsorbed in the proximal nephron by a mechanism involving endocytosis at the luminal membrane, followed by proteolysis at lysosomes.

Postmortem heart blood aspirates from 24 consecutive sudden infant deaths and 23 consecutive infant deaths from other causes yielded evidence of erythrocyte transketolase activity (ETKA) in every instance. ETKA measurements from cadaver specimens have not been previously reported. ETKA values did not correlate with the interval from death to sampling or with age at death. Activity coefficients from postmortem material corresponded closely to those from previous studies on living subjects which suggests that the method of Bayoumi and Rosalki yields valid measurements at least for 77 h postmortem. Statistical comparisons of the sudden infant deaths with non-sudden infant deaths revealed no significant differences. This investigation failed to disclose evidence in favor of the hypothesis, advanced by Read, that unsuspected thiamin deficiency may explain some cases of the sudden infant death syndrome.

The kidney plays an important role in the metabolism of proteins and peptides. Current evidence indicates that only the proximal tubule possesses the mechanism for degradation or transport of these substances and reabsorption of metabolic products. Proteins and large polypeptides filtered at the glomerulus are absorbed from proximal tubular fluid by luminal endocytosis into apical vacuoles. These fuse with primary lysosomes, where hydrolysis occurs followed by diffusion of metabolites out of the cells and into the blood. Recent evidence indicates that small linear peptides are handled by a different mechanism. It is likely that small peptides are degraded at the luminal surface of the brush border of proximal tubules, which contains many hydrolytic enzymes, by the process of membrane or contact digestion with reabsorption of the breakdown products. The probable biological significance of proximal tubular mechanisms for handling of proteins and peptides are conservation of amino acids, inactivation of toxic substances, and participation in the regulation of the circulating level of protein and peptide hormones.

Renal structural changes were studied sequentially between 1 hour and 6 days in rats treated with D-serine. Extensive necrosis of proximal straight tubules was rapid in onset and was followed by complete tubular regeneration 6 days post-treatment. The apparent progression of cellular changes was initial shrinkage, followed either by swelling and loss of apical cytoplasm or immediate lysis of cytoplasmic and nuclear contents. Tubular damage left only the basement membrane as a barrier between interstitial and luminal fluids. In similarly treated rats, proteinuria and glucosuria developed at the onset of tubular necrosis and disappeared when the tubules were completely relined by epithelium suggesting that they are due to diffusion of protein and glucose from interstitium into tubular fluid across the denuded basement membranes and that epithelial cells, under normal conditions, act as a barrier to diffusion of certain substances between the interstitium and tubular fluid.

Optimum tissue levels of magnesium and potassium ions are essential to the structure and function of the myocardium. We hypothesized that an inapparent deficiency of one or both of these elements might contribute to sudden cardiac death in a susceptible person, and in this study investigated levels of these elements in the myocardium of men experiencing sudden death to determine if there was such an association. Subjects dying suddenly with ischemic heart disease had significantly lower levels of myocardial tissue magnesium and potassium than the controls (men dying of acute trauma). The four lowest potassium values were obtained for the only men in the group with a history of angina (P less than 0.0005). Three of the men with angina also had the lowest tissue magnesium levels (P less than 0.005). The intriguing finding gives support to the hypothesis.

Reabsorption and/or degradation of proteins or peptides are functions of the proximal tubule. Large polypeptides or proteins are reabsorbed by luminal endocytosis and hydrolyzed by lysosomal enzymes. Our recent studies indicate that small linear peptides are hydrolyzed at the luminal membrane, with reabsorption of metabolites. The renal transport and hydrolysis of radiolabeled Al, All, BKN, oxytocin, glucagon, insulin, and LHRH were studied. Techniques for in vivo microinfusion of surface tubules in rats, arterial infusion in filtering and nonfiltering rat kidneys in vivo, and in vitro microperfusion of isolated rabbit nephron segments were used. Reabsorption of radiolabeled material was measured and the intact peptide or its metabolites were identified and quantified in urine, renal venous blood, bathing medium, and/or collection fluid. In addition, peptides were incubated in the presence of isolated renal membrane preparations to identify a probably cellular site of hydrolysis. The findings indicate that in proximal, but not distal tubules, radiolabeled Al, All, BKN, glucagon, and LHRH are hydrolyzed by brush border enzymes at the luminal membrane, followed by reabsorption of metabolites. In addition, it was found that, similar to the small intestine, the proximal tubule reabsorbed small peptide fragments, which were further degraded intracellurarly, In vivo inhibition studies with excess peptides revealed that hydrolysis is a more specific process than studies with excess peptides revealed that hydrolysis is a more specific process than reabsorption of metabolites. Large or small, complex peptides like insulin, oxytocin, or vasopressin that contain disulfide bridges are not hydrolyzed at the luminal brush border of the proximal tubule. In vivo sequestration and slow degradation of insulin by rat tubules suggest that this peptide is reabsorbed by endocytosis and degraded in lysosomes. Thus, as the molecular complexity or weight of a peptide increases, the mechanism for renal tubular degradation, instead of depending on luminal membrane hydrolysis, may primarily involve endocytosis and lysosomal digestion. This recently described mechanism for hydrolysis and transport of small linear peptides in the proximal nephron is characterized by having a high capacity and is analogous to membrane hydrolysis described for intestinal microvilli. The process may be biologically important to (1) conserve amino acids, (2) inactivate toxic peptides, and (3) help regulate circulating levels of peptide hormones.

A method for the large-scale production of a (1 yields 3)-beta-D-glucan glucohydrolase (EC 3.2.1.58) from the culture filtrate of Basidiomycete QM806 is described. The final preparation is homogeneous by disc electrophoresis under non-dissociating and denaturing conditions, by ultracentrifugation, and by isoelectric focusing. Various physical and chemical characteristics of the enzyme have been determined, including terminal amino acid residues, extinction coefficient, and stability to pH extremes. The N-terminal amino acids are leucine and serine (Sanger's method) and the C-terminal amino acids are alanine, serine, and glycine (hydrazinolysis). pH profile studies show that no group titrating in the region 2.5-8 is directly involved with substrate binding and that a single group having a pKa of 6.5 is involved in the catalysis. Photooxidation of the enzyme caused rapid inactivation. The pH-dependence of this photooxidation, and amino acid analysis of the photooxidized enzyme, indicate that decomposition of histidine is probably responsible for the loss of activity. Other chemical modifications performed were: treatment with hydrogen peroxide under acidic conditions, esterification with diphenyldiazomethane, and oxidation with N-bromosuccinimide. Oxidation with N-bromosuccinimide indicated that a tryptophan side-chain is involved in, but not necessary for, the catalytic activity.