Adherence to Healthy Eating Index-2015 and severity of disease in hospitalised military patients with COVID-19: a cross sectional study.

INTRODUCTION: Proper nutrition can play an important role in preventing and improving disease progression in patients with COVID-19. The Healthy Eating Index-2015 (HEI-2015) is one of the most common measures used to assess overall nutritional quality. This research aimed to identify the relationship between the HEI-2015 score and disease severity in hospitalised military patients with COVID-19. METHODS: This cross-sectional study was conducted in 295 hospitalised military patients (retired military and military reserve) with COVID-19. A validated food frequency questionnaire was used to assess food intake. To evaluate the quality of the diet, the HEI-2015 score was calculated. A multiple logistic regression analysis was performed to measure the association between HEI-2015 scores and disease severity (intensive care unit (ICU) admission and length of hospital stay greater than 4 days) in hospitalised military patients with COVID-19. RESULTS: The mean HEI-2015 score was significantly higher in non-ICU patients than in ICU patients (58.39±15.02 vs 53.54±15.65, p=0.01). After adjusting for possible confounding factors including age, sex, comorbidities, calorie intake, body mass index and physical activity, adherence to HEI-2015 inversely related to ICU admission (OR 0.98; 95% CI 0.95 to 1.00) and length of hospital stay of more than 4 days (OR 0.99; 95% CI 0.97 to 1.00) in hospitalised military patients with COVID-19, although statistically not significant. CONCLUSIONS: According to the results of the study, adherence to HEI-2015 inversely related to both ICU admission and length of hospital stay in hospitalised military patients with COVID-19, although it was not statistically significant.

Attenuation of 7-ketocholesterol- and 7ß-hydroxycholesterol-induced oxiapoptophagy by nutrients, synthetic molecules and oils: Potential for the prevention of age-related diseases.

Age-related diseases for which there are no effective treatments include cardiovascular diseases; neurodegenerative diseases such as Alzheimer's disease; eye disorders such as cataract and age-related macular degeneration; and, more recently, Severe Acute Respiratory Syndrome (SARS-CoV-2). These diseases are associated with plasma and/or tissue increases in cholesterol derivatives mainly formed by auto-oxidation: 7-ketocholesterol, also known as 7-oxo-cholesterol, and 7ß-hydroxycholesterol. The formation of these oxysterols can be considered as a consequence of mitochondrial and peroxisomal dysfunction, leading to increased in oxidative stress, which is accentuated with age. 7-ketocholesterol and 7ß-hydroxycholesterol cause a specific form of cytotoxic activity defined as oxiapoptophagy, including oxidative stress and induction of death by apoptosis associated with autophagic criteria. Oxiaptophagy is associated with organelle dysfunction and in particular with mitochondrial and peroxisomal alterations involved in the induction of cell death and in the rupture of redox balance. As the criteria characterizing 7-ketocholesterol- and 7ß-hydroxycholesterol-induced cytotoxicity are often simultaneously observed in major age-related diseases (cardiovascular diseases, age-related macular degeneration, Alzheimer's disease) the involvement of these oxysterols in the pathophysiology of the latter seems increasingly likely. It is therefore important to better understand the signalling pathways associated with the toxicity of 7-ketocholesterol and 7ß-hydroxycholesterol in order to identify pharmacological targets, nutrients and synthetic molecules attenuating or inhibiting the cytotoxic activities of these oxysterols. Numerous natural cytoprotective compounds have been identified: vitamins, fatty acids, polyphenols, terpenes, vegetal pigments, antioxidants, mixtures of compounds (oils, plant extracts) and bacterial enzymes. However, few synthetic molecules are able to prevent 7-ketocholesterol- and/or 7ß-hydroxycholesterol-induced cytotoxicity: dimethyl fumarate, monomethyl fumarate, the tyrosine kinase inhibitor AG126, memantine, simvastatine, Trolox, dimethylsufoxide, mangafodipir and mitochondrial permeability transition pore (MPTP) inhibitors. The effectiveness of these compounds, several of which are already in use in humans, makes it possible to consider using them for the treatment of certain age-related diseases associated with increased plasma and/or tissue levels of 7-ketocholesterol and/or 7ß-hydroxycholesterol.

Towards Higher Oil Yield and Quality of Essential Oil Extracted from Aquilaria malaccensis Wood via the Subcritical Technique.

A method that delivers a high yield and excellent quality of essential oil, which retains most of its value-added compounds, and undergoes least change after the extraction process, is greatly sought after. Although chemical free methods are acceptable, they call for an extensive processing time, while the yield and quality from these methods are often disappointing. This work utilizes subcritical water technology to address these issues. In this undertaking, essential oil was extracted from Aquilaria malaccensis wood by way of subcritical conditions, and characterized through gas chromatography/mass spectroscopy (GC/MS). Optimization through response surface methodology revealed temperature to be the most critical factor for the extraction process, while the optimum conditions for temperature, sample-to-solvent ratio, and time for subcritical water extraction was revealed as 225 °C, 0.2 gr/mL, and 17 min, respectively. The subcritical water extraction technique involves two simultaneous processes, which are based on good fitting to the two-site kinetic and second order model. In comparison to the hydrodistillation method, GC/MS results indicated that the quality of A. malaccensis' wood oils, derived through the subcritical water technique, are of significantly better quality, while containing many constructive value-added compounds, such as furfural and guaiacol, which are useful for the production of pesticides and medicines. Pore size, functional groups, and morphology analysis revealed the occurrence of substantial damage to the samples, which facilitated an improved extraction of bio-products. In comparison to conventional methods, the use of the subcritical method not only involves a shorter processing time, but also delivers a higher oil yield and quality.

Induction of a non-apoptotic mode of cell death associated with autophagic characteristics with steroidal maleic anhydrides and 7ß-hydroxycholesterol on glioma cells.

Steroidal maleic anhydrides were prepared in one step: lithocholic, chenodeoxicholic, deoxicholic, ursocholic, and hyodeoxicholic acid derivatives. Their capability to induce cell death was studied on C6 rat glioma cells, and 7ß-hydroxycholesterol was used as positive cytotoxic control. The highest cytotoxicity was observed with lithocholic and chenodeoxicholic acid derivatives (23-(4-methylfuran-2,5-dione)-3α-hydroxy-24-nor-5ß-cholane (compound 1a), and 23-(4-methylfuran-2,5-dione)-3α,7α-dihydroxy-24-nor-5ß-cholane (compound 1b), respectively), which induce a non-apoptotic mode of cell death associated with mitochondrial membrane potential loss and reactive oxygen species overproduction. No cells with condensed and/or fragmented nuclei, no PARP degradation and no cleaved-caspase-3, which are apoptotic criteria, were observed. Similar effects were found with 7ß-hydroxycholesterol. The cell clonogenic survival assay showed that compound 1b was more cytotoxic than compound 1a and 7ß-hydroxycholesterol. Compound 1b and 7ß-hydroxycholesterol also induce cell cycle modifications. In addition, compounds 1a and 1b, and 7ß-hydroxycholesterol favour the formation of large acidic vacuoles revealed by staining with acridine orange and monodansylcadaverine evocating autophagic vacuoles; they also induce an increased ratio of [LC3-II / LC3-I], and modify the expression of mTOR, Beclin-1, Atg12, and Atg5-Atg12 which is are autophagic criteria. The ratio [LC3-II / LC3-I] is also strongly modified by bafilomycin acting on the autophagic flux. Rapamycin, an autophagic inducer, and 3-methyladenine, an autophagic inhibitor, reduce and increase 7ß-hydroxycholesterol-induced cell death, respectively, supporting that 7ß-hydroxycholesterol induces survival autophagy. Alpha-tocopherol also strongly attenuates 7ß-hydroxycholesterol-induced cell death. However, rapamycin, 3-methyladenine, and α-tocopherol have no effect on compounds 1a and 1b-induced cell death. It is concluded that these compounds trigger a non apoptotic mode of cell death, involving the mitochondria and associated with several characteristics of autophagy.

Analysis of the association between IL-23R rs11209026 polymorphism and incidence of atherosclerosis.

Th17/IL-23 axis is an important pro-inflammatory pathway in atherosclerosis. IL-23 receptor (IL-23R) pathway has an important role in T-helper cells 17 (Th17) differentiation and survival. We compared normal subjects and patients with atherosclerosis in terms of the R381Q variant of the IL-23R gene as a functional single-nucleotide polymorphism (SNP). This case-control study recruited 200 patients who presented with cardiovascular symptoms to Afshar Hospital, Yazd, Iran. The participants were allocated to five groups based on angiographic results. The severity of the disease was determined according to the numbers of involved vessels. Patients with normal coronary arteries, minimal coronary artery involvement, one involved vessel, two involved vessels and three-vessel disease were allocated to groups I-V, respectively. DNA was extracted from whole blood samples by the salting-out method. Genotyping was performed by polymerase chain reaction followed by restriction fragment length polymorphism assay and multinomial logistic regression for analyses The presence of SNP A>G rs11209026 of IL-23 receptor gene was significantly associated with disease severity (P = 0.008). The frequencies of the heterozygous (AG) genotype in the control group and subjects with minimal involvement, and patients with one-, two-, and three-vessel disease were 22.5%, 12.5%, 10%, 10.24% and 4.8%, respectively. Our results indicated an association between the rs11209026 G>A polymorphism of the IL-23 receptor gene and the risk of atherosclerosis. This genetic variant may in fact cause protection against atherosclerosis progression. However, further studies on gene polymorphism and cell expression are required to clarify the mechanisms involved in the pathogenesis of atherosclerosis.

TCN 201 selectively blocks GluN2A-containing NMDARs in a GluN1 co-agonist dependent but non-competitive manner.

Antagonists that are sufficiently selective to preferentially block GluN2A-containing N-methyl-d-aspartate receptors (NMDARs) over GluN2B-containing NMDARs are few in number. In this study we describe a pharmacological characterization of 3-chloro-4-fluoro-N-[4-[[2-(phenylcarbonyl)hydrazino]carbonyl]benzyl]benzenesulphonamide (TCN 201), a sulphonamide derivative, that was recently identified from a high-throughput screen as a potential GluN2A-selective antagonist. Using two-electrode voltage-clamp (TEVC) recordings of NMDAR currents from Xenopus laevis oocytes expressing either GluN1/GluN2A or GluN1/GluN2B NMDARs we demonstrate the selective antagonism by TCN 201 of GluN2A-containing NMDARs. The degree of inhibition produced by TCN 201 is dependent on the concentration of the GluN1-site co-agonist, glycine (or D-serine), and is independent of the glutamate concentration. This GluN1 agonist-dependency is similar to that observed for a related GluN2A-selective antagonist, N-(cyclohexylmethyl)-2-[{5-[(phenylmethyl)amino]-1,3,4-thiadiazol-2-yl}thio]acetamide (TCN 213). Schild analysis of TCN 201 antagonism indicates that it acts in a non-competitive manner but its equilibrium constant at GluN1/GluN2A NMDARs indicates TCN 201 is around 30-times more potent than TCN 213. In cortical neurones TCN 201 shows only modest antagonism of NMDAR-mediated currents recorded from young (DIV 9-10) neurones where GluN2B expression predominates. In older cultures (DIV 15-18) or in cultures where GluN2A subunits have been over-expressed TCN 201 gives a strong block that is negatively correlated with the degree of block produced by the GluN2B-selective antagonist, ifenprodil. Nevertheless, while TCN 201 is a potent antagonist it must be borne in mind that its ability to block GluN2A-containing NMDARs is dependent on the GluN1-agonist concentration and is limited by its low solubility.

Regional- and agonist-dependent facilitation of human neurogastrointestinal functions by motilin receptor agonists.

BACKGROUND AND PURPOSE: Delayed gastric emptying is poorly managed. Motilin agonists are potential treatments but inadequate understanding into how enteric nerve functions are stimulated compromises drug/dose selection. Resolution is hampered by extreme species dependency so methods were developed to study human gastrointestinal neuromuscular activities and the neurobiology of motilin. EXPERIMENTAL APPROACH: Protocols to study neuromuscular activities were developed for different regions of human stomach and intestine (71 patients) using circular muscle preparations and electrical field stimulation (EFS) of intrinsic nerves. Other tissues were fixed for immunohistochemistry. KEY RESULTS: EFS evoked contractions and/or relaxations via cholinergic and nitrergic neurons, with additional tachykinergic activity in colon; these were consistent after 154 min (longer if stored overnight). Motilin 1-300 nM and the selective motilin agonist GSK962040 0.1-30 µM acted pre-junctionally to strongly facilitate cholinergic contractions of the antrum (E(max) ≈ 1000% for motilin), with smaller increases in fundus, duodenum and ileum; high concentrations increased baseline muscle tension in fundus and small intestine. There were minimal effects in the colon. In the antrum, cholinergic facilitation by motilin faded irregularly, even with peptidase inhibitors, whereas facilitation by GSK962040 was long lasting. Motilin receptor immunoreactivity was identified in muscle and myenteric plexus predominantly in the upper gut, co-expressed with choline acetyltransferase in neurons. CONCLUSIONS AND IMPLICATIONS: Motilin and GSK962040 strongly facilitated cholinergic activity in the antrum, with lower activity in fundus and small intestine only. Facilitation by motilin was short lived, consistent with participation in migrating motor complexes. Long-lasting facilitation by GSK962040 suggests different receptor interactions and potential for clinical evaluation.

Photocatalytic reduction of Cr(VI) and Ni(II) in aqueous solution by synthesized nanoparticle ZnO under ultraviolet light irradiation: a kinetic study.

Photocatalytic removal of Cr(VI) and Ni(II) from aqueous solution using synthesized nanoparticle ZnO under ultraviolet (UV) light irradiation was studied in this work. Firstly, nanoparticle ZnO was prepared by the chemical method with an organic chemical inhibitor. Then removal efficiency of Cr(VI) and Ni(II) by nanoparticle ZnO was investigated with variation of the solution pH, ZnO dosage, contact time and initial Cr(VI) and Ni(II) concentration. Maximum removal of Cr(VI) and Ni(II) was observed at near-neutral pH because the reduced photocatalytic activity of ZnO at exceedingly low and high pH values originates from either acidic/photochemical corrosion of the catalyst and/or surface passivation with Zn(OH)2. As the ZnO dosage increased, the removal efficiency of Cr(VI) and Ni(II) was continuously enhanced, but was gradually decreased above 1.25 g/l due to the increased blockage of the incident UV light used for the photocatalytic reaction. The optimum ZnO dosage was determined as 1 g/l. Removal efficiencies of Cr(VI) and Ni(II) decreased as initial Cr(VI) and Ni(II) concentration increased, due to an increased inhibition effect on the surface of ZnO resulting from the decreased reaction sites on the surface of ZnO required for the further photocatalytic reaction.

Adsorption of chromium (VI) from aqueous solution by sugar beet bagasse-based activated charcoal.

Chromium (VI) is known to be potentially carcinogenic and mutagenic in humans. A low-cost industrial solid bioadsorbent, bagasse-based activated charcoal (BAC), has been investigated for removal of chromium from aqueous solution. All the experiments were carried out in batch process with laboratory-prepared samples to study the effects of adsorbent dose, contact time, pH and initial concentration of Cr(VI). The removal of chromium ion was found to be highly dependent on the pH of the solution, adsorbent dose and contact time. Also the equilibrium adsorption was analyzed by the Freundlich and Langmuir isotherm models. It was found that the Freundlich isotherm model best described the sorption of chromium by sugar beet bagasse-based activated charcoal (r2 > 0.9927). Experimental data of kinetic studies were fitted to pseudo-first-order, pseudo-second-order and modified pseudo-first-order models. The results showed pseudo-second order kinetics was best fitted to the collected data (r2 > 0.9893). Optimum conditions for adsorption were determined at pH 2 and a contact time of 180 minutes (92.7% removal). These retention capacities suggest that BAC can provide a simple, effective, and cheap method for removing Cr(VI) ions from effluents and water resources.

Study on bone age in pediatric patients with congenital heart disease and its relation with cyanosis and pulmonary artery pressure.

The aim of this study is to evaluate the growth failure in children with Congenital Heart Diseases (CHD) associated with the Pulmonary Hypertension (PH) and cyanosis. Growth parameters including weight, height and head circumference of 120 cases with congenital heart defects aged 6 months to 14 years were compared with standard growth curves (50th percentile) between November 2007 and November, 2008. Of all, sixty five (54.1%) were male and 55 (45.8%) were female. The patients were classified into four groups based on the presence or absence of PH and cyanosis. The gap between chronological age and bone age (BA) for all subjects was determined. Growth disturbance in weight, height and head circumference was detected in 80 (66.7%), 79 (65.8%) and 41 (34.2%) of the patients, respectively. Bone age delay was seen in fifty five percent of the cases. Generally, delay in all parameters was more seen in acyanotic patients with pulmonary hypertension. In subjects with cyanosis whether in addition to PH or not, bone age was significantly retarded. Etiology of growth failure in children with CHD is multifactorial. Further studies are required to assess the role of different factors in this field.

Cardiac involvement of major thalassemia and evaluation of total serum creatine kinase and creatine kinase-mB isoenzyme and cardiac troponinI in these patients.

The goal of this study was the evaluation of specific markers of myocardial injury that includes CK-MB and troponin I in major thalassemic patients. Regular blood transfusion is the main treatment in major thalassemia. One of the most important complications of regular blood transfusion is iron overload that eventually involves many organs like heart and cause myocardial injury. Sixty patients with transfusion-dependent major thalassemia, at the age range of 8 to 15 years in Tabriz Pediatric Medical Center were chosen. Measurement of Hb, Hct and serum ferritin were performed in hospital laboratory, but total serum Creatine Kinase (CK) by photometric and isoenzyme of CK-MB by immunologic DGKC and cardiac troponin I (cTnI) were tested by ELISA methods in Shaheed Madani heart center laboratory before blood transfusion. For all patients echocardiography and ECG assessment of cardiac function were done by a pediatric cardiologist and results were statistically analyzed. Forty nine patients (group A) had normal left ventricular ejection fraction (LVEF = 50-70%) and 11 patients (group B) had reduced LVEF (20-45%). There was no statistical difference between two groups in average volume of blood transfusion (p = 0.074). Although total CK and CK-MB isoenzyme were higher in group B but there was no statistically meaningful difference between two groups (p = 0.123, p = 0.111). Troponin I also was higher in group B but statistically analysis showed no correlation between cardiac function and troponin I level in these groups (p = 0.827). This study showed that cardiac markers are not helpful for recognition of cardiac involvement in major thalassemia.

GB virus type C infection in hemodialysis patients considering co-infection with hepatitis C virus.

GB virus type C is a well-known viral agent with capability of infecting patients undergoing hemodialysis. Liver enzyme levels in infected individuals have been reported to remain within the normal range. Simultaneous infection of GBV-C and other viral agents may occur due to common routes of transmission. A total of 104 hemodialysis patients living in Tehran were included in this case-control study (53 patients with HCV infection, group I; and 51 with no HCV infection, group II). Diagnosis was made by detection Anti-E(2) protein using ELISA and HCV-RNA using RT-PCR. History of HBV-infection, organ transplantation, depression, malignancies, chemotherapy, diabetes mellitus, thyroid disorders and chronic cutaneous disorders were considered. Patients were evaluated for high- risk behaviors such as intravenous drug injection, addiction or substance abuse. A total of 14 patients (13.6%) were GBV-C-infected. Four of them were co-infected with HCV. All patients with GBV-C infection had viral genotype 2. Thirteen patients (12%) had a history of multiple blood transfusions. Mean (+/-SD) age of GBV-C-infected patients was 48.7+/-13.8 years. Among GBV-C infected patients, three patients had a history of organ transplantation and three had a co-morbidity of diabetes mellitus. This study as the first case-control study to evaluate the association between GBV-C and HCV infection, to our knowledge, shows hemodialysis patients living in Tehran are infected with GBV-C with intermediate level of frequency. The association of GBV-C transmission with other viral blood-borne agents might be necessary.

Analysis of class-switched memory B cells in patients with common variable immunodeficiency and its clinical implications.

BACKGROUND: Common variable immunodeficiency (CVID) comprises a heterogeneous group of primary immunodeficiency disorders characterized by hypogammaglobulinemia leading to recurrent infections. Some patients with CVID are more susceptible to earlier onset of respiratory disease and bronchiectasis. It has been suggested that memory B cells, characterized by CD27 expression, can be used as a means to classify subsets of CVID patients. OBJECTIVE: The aim of this study was to classify a sample of Iranian patients with CVID by quantification of peripheral blood memory B cells and immature B cells and to assess the relationship between this classification and the clinical characteristics of the patients. METHODS: The study included 29 patients with CVID and 20 healthy controls. Patients were grouped as follows, according to the quantification of peripheral memory B cells: group I had less than 0.4% switched memory B cells (CD27+, immunoglobulin [Ig] M-, IgD-) in peripheral blood lymphocytes (PBL), while in group II switched memory B cells represented more than 0.4% of PBL. Group I patients were further subdivided into groups Ia and Ib according to the proportion of CD21- peripheral B cells. The clinical and laboratory findings for the patients were then compared among the 3 groups. RESULTS: The percentage of switched memory B cells (CD27+IgM-IgD- cells in peripheral B lymphocytes) was markedly reduced in CVID patients compared with controls (P < .001). This percentage was less than 0.4% (group I) in 20 patients (69%) (P < .05). In the remaining 9 patients (group II) and all healthy controls, the percentage was greater than 0.4%. Bronchiectasis was more frequent in group I than group II (P < .05). Following subdivision of group I patients into groups Ia and Ib based on CD21 peripheral B cells, the rate of autoimmunity was found to be much higher in group Ia than group Ib. CONCLUSIONS: CVID patients with reduced numbers of switched memory B cells are more prone to recurrent respiratory infections and development of bronchiectasis, and as such, need more special care than other CVID patients. Thus, classification of CVID patients by assessment of switched memory B cells could help physicians to predict clinical prognosis of these patients.

Expression of neuronal nitric oxide synthase during embryonic development of the rat optic vesicle.

The expression of neuronal nitric oxide synthase during the development of rat optic vesicle from embryonic day E14 to E18 was analyzed by histochemical procedures. The samples were frozen and cut on a cryostat and then studied by using the light microscope. Expression of nNOS was first seen on E14 in cells of Cajal-Retzius located in the marginal zone of optic vesicle. NADPH-d persisted in this layer throughout the embryonic period and began to decrease on E20. At E16, the optic vesicle has four NADPH-d positive layers. At E18, NADPH-d reactivity observed at low magnification showed five clearly defined layers. In the late stages, the most notable feature was a decrease in histochemical reaction of the marginal zone and at these stages, the layer IV showed less staining than the rest of the cortical plate. The observations suggest that nitric oxide is synthesized during embryonic life processes and this is related to maturational processes.

Total synthesis and determination of the absolute configuration of coscinosulfate. A new selective inhibitor of Cdc25 protein phosphatase.

The first total synthesis of coscinosulfate 1, a metabolite isolated from a sea sponge, starting from (+)-sclareolide 3 is described. The convergent synthesis strategy relies on the coupling of sulfone 21 with the bromide 26. The sulfone fragment 21 was obtained by successive asymmetric aldol reaction with aldehyde 2 to introduce the stereocenters at C-12 and C-13, followed by one-carbon homologation via Horner-Wadsworth-Emmons olefination. The selective sulfatation at C-12 was accomplished through the quinone intermediate 31 obtained by selective oxidation of hydroquinone 30; this, when followed by reduction, furnished the desired coscinosulfate 1. X-ray analysis of the intermediate aldehyde 18 confirmed the proposed structure.

Coscinosulfate, a CDC25 phosphatase inhibitor from the sponge Coscinoderma mathewsi.

The dual specificity CDC25 phosphatases dephosphorylate two inhibitory phospho-amino acids of cyclin-dependent kinases, a major family of cell cycle regulators. CDC25 inhibitors constitute new anti-mitotic agents with potential anticancer activity. While screening through a collection of natural products derived from marine organisms for CDC25A inhibitors, we purified and identified coscinosulfate 1, a sesquiterpene sulfate from the New Caledonian sponge Coscinoderma matthewsi, along with 4. The purified compound 1 displayed significant inhibitory activity towards CDC25A (IC(50): 3 microM).

Total synthesis of uracil analogues of sinefungin.

Analogues of sinefungin derivatives 18a and 18b have been prepared from uridine and L-aspartic acid. The key step in the synthesis was the coupling of the radical derived from 14 with the unsaturated amide 13. The latter was produced from the known N-hydroxy-2-thiopyridone ester of L-aspartic acid 12 with the olefin 11. Thus, the essential carbon skeleton was constructed by way of two radical coupling reactions. These analogues as well as 1a and 1b synthesized previously were tested for their antileishmanial effect in vivo and for their inhibitory activity of protein carboxymethylase (protein methylase II). The replacement of the adenine moiety by uracil or dihydrouracil considerably decreases the antiparasitic activity and the affinity for protein methylase II. The synthetic (S)-sinefungin was as active as the natural one. Interestingly, the C-6' epimer 1b was 50% less active in vitro than the natural sinefungin, but both had identical affinities for the target enzyme.

beta-Glucosidase isoenzymes in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and patients with type 1 Gaucher disease.

Lymphoid cell lines (LCL) from 3 adult patients with non-neuropathic Gaucher disease were established by Epstein-Barr virus (EBV) transformation and were investigated from the view of enzymology. Glucosylceramide-beta-glucosidase (GlcCer-beta-glucosidase) was present in soluble and particulate fraction of LCL from normal subjects and was deficient in type 1 Gaucher LCL; the deficiency of all molecular forms, shown by electrofocusing, indicates that they are coded by the same gene. The existence of two non-specific beta-glucosidases, one soluble (minor), the other membrane-bound (major), was demonstrated in leucocytes and LCL from normals; in Gaucher LCL, these were also present in a normal range. Characteristic properties of the non-specific membrane-bound beta-glucosidase were defined: lability at acidic pH and strong inhibitory effect by detergents. These properties allowed to discriminate it from the lysosomal GlcCer-beta-glucosidase and to define optimal assay conditions for determination of residual GlcCer-beta-glucosidase activity in Gaucher disease, using artificial substrate, without interference of non-specific membrane-bound beta-glucosidase. These results demonstrate that EBV-transformed LCL represent an accurate model system for enzymatic studies of Gaucher disease.