Adjuvant use of melatonin for relieving symptoms of painful diabetic neuropathy: results of a randomized, double-blinded, controlled trial.

PURPOSE: The trial aimed to investigate the effectiveness of exogenous melatonin as an adjuvant to pregabalin for relief of pain in patients suffering from painful diabetic neuropathy (PDN). PATIENTS AND METHODS: This randomized, double-blind, placebo-controlled trial was carried out between October 2019 and December 2020 in an outpatient specialty clinic in Iran. One-hundred-three type 2 diabetic patients suffering from PDN were randomized into either the melatonin group (n = 52) or the placebo group (n = 51). Besides pregabalin at a dose of 150 mg per day, patients started with melatonin or an identical placebo, at a dose of 3 mg/day at bedtime for 1 week, which was augmented to 6 mg/day for further 7 weeks. The primary outcomes were changes in mean NRS (numerical rating scale) pain score from baseline to endpoint and responder rate (patients with a reduction of 50% and higher in average pain score compared with baseline). Secondary endpoints were changes in mean NRS pain-related sleep-interference score, overall improvement evaluated by Patient and Clinical Global Impressions of Change (PGIC, CGIC), and impact of the intervention on patient's Health-related quality of life (QOL). All analyses were conducted on an Intention-to-Treat (ITT) analysis data set. RESULTS: At the study endpoint, treatment with melatonin resulted in a considerably higher reduction in the mean NRS pain score in comparison with placebo (4.2 ± 1.83 vs. 2.9 ± 1.56; P-value < 0.001). In terms of treatment responders, a greater proportion of melatonin-treated patients satisfied the responder criterion than placebo-treated patients (63.5% vs. 43.1%). Melatonin also reduced pain-related sleep interference scores more than did placebo (3.38 ± 1.49 vs. 2.25 ± 1.26; P-value < 0.001). Further, at the endpoint, more improvement was also seen in terms of PGIC, CGIC, and Health-related QOL in patients treated with melatonin than placebo. Melatonin was also well tolerated. CONCLUSION: The present results showed that melatonin as an adjunct therapy to pregabalin might be helpful for use in patients with PDN. However, confirmation of these results requires further studies.

Effects of dietary yeast cell wall on biochemical indices, serum and skin mucus immune responses, oxidative status and resistance against Aeromonas hydrophila in juvenile Persian sturgeon (Acipenser persicus).

The present study aims to shed light on the effects of yeast cell wall (ImmunoWall®) supplementation on biochemical indices, oxidative status, serum and mucus immune responses as well as disease resistance of juvenile Persian sturgeon (Acipenser persicus). For this purpose, one hundred fifty three juvenile Persian sturgeons (47.78 ± 0.39 g) were distributed into nine tanks (500 L) and fed with basal diets containing two levels of yeast cell wall (YCW) 0.5% (T1) and 1% (T2) and a diet without YCW as control (0%). As shown by the results obtained at the end of 56-day feeding trial, YCW had no significant effect on glucose, cortisol, SGOT, lysozyme and IgM in serum (P > 0.05) albeit an enhancement of cholesterol, LDH, ALP and SOD and ACH50 was observed in fish fed YCW supplemented diets. However, plasma triglyceride levels were lower in fish fed YCW compared with the control group. Also, total protein content, lysozyme and protease activities in skin mucus were unaffected by the supplemented diets (P > 0.05) and only total immunoglobulin and ALP enzyme activity were significantly increased in T1 and T2 groups (P > 0.05). The cumulative mortality of the fish fed supplemented diets at the end of disease challenge was 100% where cumulative mortality of those fed the control diet was 75% (P < 0.05). The present study shows that increasing immune parameters in serum and mucus of juvenile Persian sturgeon by YCW dietary supplementation did not improve resistance against Aeromonas hydrophila. According to the obtained results, the YCW supplementation at 0.5 and 1% in the juvenile Persian sturgeon diet is not recommended.

Discovery of phthalimide derivatives as novel inhibitors of a soluble epoxide hydrolase.

Soluble epoxide hydrolase (sEH) inhibitors are effective in reducing blood pressure, inflammation, and pain in a number of mammalian disease models. As most classical urea-based sEH inhibitors suffer from poor solubility and pharmacokinetic properties, the development of novel sEH inhibitors with an improved pharmacokinetic specification has received a great deal of attention. In this study, a series of amide-based sEH inhibitors bearing a phthalimide ring as the novel secondary pharmacophore (P2 ) was designed, synthesized, and evaluated. Docking results illustrated that the amide group as the primary pharmacophore (P1 ) was placed at a suitable distance from the three key amino acids (Tyr383, Tyr466, and Asp335) for an effective hydrogen bonding. In agreement with these findings, most of the newly synthesized compounds demonstrated moderate to high sEH inhibitory activities, relative to 12-(3-adamantan-1-yl-ureido)dodecanoic acid as the reference standard. Compound 12e with a 4-methoxybenzoyl substituent exhibited the highest sEH inhibitory activity, with an IC50 value of 1.06 nM. Moreover, the ADME properties of the compounds were evaluated in silico, and the results revealed appropriate predictions.

Very rapid amyloid fibril formation by a bacterial lipase in the absence of a detectable lag phase.

The conversion of proteins from their soluble states into well-organized amyloid fibrils has received abundant attention. This process typically consists of three stages: lag, growth and plateau phases. In this study, the process of amyloid fibril formation by lipase from Pseudomonas sp. after diluting out urea was examined by Thioflavin T (ThT) fluorescence, Congo red (CR) binding, 8-anilinonaphthalene-1-sulfonic acid (ANS) binding, dynamic light scattering (DLS), circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopies, X-ray diffraction (XRD) and transmission electron microscopy (TEM). To exclude the presence of preformed aggregates in the pure lipase sample, aforementioned assays were also performed for the protein unfolded in urea before dilution. The aggregates formed immediately after dilution were found to bind to ThT and CR and contain a significant amount of ß-sheet structure, as determined by far-UV CD and FTIR spectroscopies, as well as XRD analysis. Moreover, these aggregates present, at least in part, a fibrillar morphology, as deduced with TEM. This examination showed that lipase fibril formation proceeds quickly after dilution, within a few seconds, without a detectable lag phase. We also investigated bacterial inclusion bodies formed after expression of lipase in E. coli, providing evidence for the existence of rapidly formed amyloid-like structural and tinctorial properties in the lipase-containing inclusion bodies.

Down syndrome and consanguinity.

BACKGROUND: Among the genetics disorders, Down syndrome (DS) is the major cause of mental retardation, congenital heart and intestinal disease. So far, no certain therapeutic method has been suggested for the treatment of this syndrome. The aim of the current survey was to investigate the frequency of parental consanguinity, maternal age in the patients with DS. MATERIALS AND METHODS: This study was conducted on 38 consecutive patients with clinically and laboratory confirmed DS who referred to the genetic lab of a referral University Hospital. The G-banding method for karyotyping was employed. RESULTS: The patients were 21 males and 17 females within the age of 16 days to 28 years old. Free trisomy (92.1%, n = 35) was the most common chromosomal abnormality. The frequency of DS was higher among the non-consanguine marriages (71.1%) in comparison with the consanguine marriages (28.9%). Mean age of the mothers in the consanguine marriages (mean = 27.1 ± 6.3) was lower than in the non-consanguine marriages (mean = 31.1 ± 7.7). CONCLUSION: Higher frequency of DS among the non-consanguine marriages in comparison with the consanguine marriages, may suggest that DS diagnostic tests might be done on all embryos regardless of the parents' familial relationship.

Effects of leukocyte-platelet-rich fibrin and advanced platelet-rich fibrin on the viability and migration of human gingival fibroblasts.

BACKGROUND: Platelet products play a fundamental role in the process of healing. The new generation of platelet-rich fibrin (PRF), namely advanced PRF (A-PRF), has different biological and mechanical properties compared to those of leukocyte-PRF (L-PRF). This study aimed to compare the effects of L-PRF and A-PRF on the viability and migration of human gingival fibroblasts (HGFs). MATERIALS AND METHODS: In this in vitro study, the effects of A-PRF and L-PRF on the viability and migration of HGFs after 24 and 48 h were evaluated using the methyl thiazolyl tetrazolium assay. The viability of the negative control culture medium was considered to be 100%. The mean optical density of the test groups was divided by that of the negative control group and reported as percentage. One-way ANOVA was applied to assess the effects of time and type of PRF on the viability and migration of HGFs. Pairwise comparisons were made using the Tukey's test. RESULTS: At 24 h, cell viability in the L-PRF group was significantly higher than that in the A-PRF group (P < 0.05). However, no significant difference was noted between the two groups at 48 h. At 24 h, L-PRF caused significantly higher cell migration compared to the negative control group, whereas at 48 h, both A-PRF and L-PRF significantly increased cell migration compared to the control group. CONCLUSION: Within the limitations of this study, L-PRF and A-PRF had significant effects on the viability and migration of HGFs. Further studies on these platelet concentrates are warranted.

Prevalence of ABO and Rh blood groups and their association with demographic and anthropometric factors in an Iranian population: Mashad study.

BACKGROUND: Blood groups appear to be markers for various human diseases and their distribution among different communities, ethnic groups and geographical boundaries varies over time. AIMS: We aimed to investigate the frequency of ABO and Rh blood groups and their relationship with demographic and anthropometric characteristics among Iranian residents in Mashad. METHODS: ABO and Rh blood groups were determined among 7268 participants from the MASHAD cohort study and their relationships with demographic and anthropometric parameters were evaluated. This part of the study was done in January 2017. Student t-test, ANOVA, Bonferroni' and Chi-squared were used for comparison of quantitative and qualitative variables. RESULTS: The most common blood group was O (33.8%); AB was the least common (8.3%). The prevalence of Rh-positive and Rh-negative was 88.2% and 11.8% respectively. There were statistically significant associations between ABO blood groups and demi-span (P = 0.03), even after correction for multiple comparisons. CONCLUSION: Our findings showed there was no relationship between ABO blood groups and demographic characteristics although there was an association with anthropometric measurements such as demi-span.

Modification of Sexual Hormones in Rheumatoid Arthritis Patients by M2000 (ß-D-mannuronic Acid) as a Novel NSAID with Immunosuppressive Property.

BACKGROUND: Based on in-vitro, in-vivo and human studies, the ß-D-mannuronic acid (M2000) has been introduced as a novel non-steroidal anti-inflammatory drug (NSAID) with immunosuppressive properties. OBJECTIVE: This study aimed to evaluate the efficacy of this drug on serum level of sex hormones (Estradiol, Progesterone, and DHEAS) in rheumatoid arthritis (RA) patients. METHODS: The present research was performed on 10 RA patients who had an inadequate response to conventional treatments (clinical trial identifier: IRCT2014011213739N2). During this trial, the patients were permitted to continue the conventional therapy along with adding M2000 orally at a dose of 500 mg twice daily for 12 weeks. Serum samples were collected in a normal group, patient group (at baseline) and treatment group (after 12 weeks). The samples were tested for evaluating the serum level of Estradiol, Progesterone, and DHEAS using chemiluminescent microparticle immunoassay. RESULTS: Data showed that the serum level of estradiol was reduced (both in men and women) during the treatment with M2000 (after 12 weeks), but there was no significant difference in the non-treated group with M2000 (p > 0.05). In addition, the serum level of progesterone and DHEAS significantly increased following the 12-week administration of M2000 in both male and female patients, compared to the non-treated group with M2000 (p < 0.001, p < 0.05, p < 0.05, p < 0.01, respectively). CONCLUSION: The present research showed that the sex hormones might be modified by M2000 therapy in RA patients by increasing the serum level of progesterone and DHEAS compared to healthy individuals.

Comparison of the molten globule states of thermophilic and mesophilic alpha-amylases.

In recent years great interest has been generated in the process of protein folding, and the formation of intermediates during the folding process has been proven with new experimental strategies. In the present work, we have examined the molten globule state of Bacillus licheniformis alpha-amylase (BLA) by intrinsic fluorescence and circular dichroism spectra, 1-anilino naphthalene-8-sulfonate (ANS) binding and proteolytic digestion by pepsin, for comparison to its mesophilic counterpart, Bacillus amyloliquefaciens alpha-amylase (BAA). At pH 4.0, both enzymes acquire partially folded state which show characteristics of molten globule state. They unfold in such a way that their hydrophobic surfaces are exposed to a greater extent compared to the native forms. Chemical denaturation studies by guanidine hydrochloride and proteolytic digestion with pepsin show that molten globule state of BLA is more stable than from BAA. Results from gel filtration indicate that BAA has the same compactness at pH 4.0 and 7.5. However, molten globule state of BLA is less compact than its native state. The effects of polyols such as trehalose, sorbitol and glycerol on refolding of enzymes from molten globule to native state were also studied. These polyols are effective on refolding of mesophilic alpha-amylase but only slightly effect on BLA refolding. In addition, the folding pathway and stability of intermediate state of the thermophilic and the mesophilic alpha-amylases are discussed.

Investigation of Protective Effect of Matricaria Chamomilla L. Extract on Methotrexate-Induced Hepatotoxicity in Wistar Rat

Abstract Methotrexate (MTX) was shown to cause oxidative stress and liver damage. The objective was to investigate the possible protective effects of Matricaria Chamomilla L. (chamomile) extract with anti-oxidant and anti-inflammatory properties on the methotrexate-induced liver toxicity. Twenty four Wistar rats were divided into four groups. MTX group was injected intraperitoneally on days 7 and 14 with 20 mg/kg methotrexate. Groups CE200 (chamomile extract 200 mg/kg/day) and CE300 (chamomile extract 300 mg/kg/day) received the same dose of methotrexate added with chamomile extract orally for 15 days at 200 mg/kg and 300 mg/kg respectively and the last group was healthy control group. Results of biochemical analyses indicated serum liver biomarkers (aminotransferases), alkaline phosphatase (ALP), albumin, and liver content of anti-oxidant enzymes (catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px)), reduced glutathione (GSH) and total anti-oxidant capacity (TAC) significantly increased (P <0.05-0.001) to normal in the CE treated groups compared to those of the MTX group. Serum bilirubin and hepatic malondialdehyde (MDA) levels significantly increased (P ˂0.001) in MTX group compared to those of the control group and decreased in CE200 and CE300 groups compared to those of the MTX group. Histopathological study showed inflammatory damage, necrotic cells and lipid infiltration in MTX group. In the groups treated with the chamomile extract, a significant improvement was observed in liver tissue in response to increased dose of the extract. In conclusion, chamomile extract administration could have a protective role in methotrexate-induced liver toxicity in rats through improving anti-oxidant defense system.

Hydrophobic substitution of surface residues affects lipase stability in organic solvents.

A novel lipase has been recently isolated from a local Pseudomonas sp. (GQ243724). In the present study, we have tried to increase the organic solvent stability of this lipase using site-directed mutagenesis. Eight variants N219L, N219I, N219P, N219A, N219R, N219D, S251L, and S251K were designed to change the surface hydrophobicity of this enzyme with respect to the wild-type. Among these variants, the stability of N219L and N219I significantly increased in the presence of all tested organic solvents, whereas two mutants (N219R and N219D) significantly exhibited decreased stabilities in all the organic solvent studied, suggesting that improvement of hydrophobic patches on the enzyme surface enhances the stability in organic media. Furthermore, replacing Ser²5¹ with hydrophobic residues on the enzyme surface dramatically diminished its stability in the tested condition. In spite of the distance of the mutated sites from the active site, the values of k(cat) and K(m) were affected. Finally, structural analysis of the wild-type and mutated variants was carried out in the presence and absence of some organic solvents using circular dichroism and fluorescence spectroscopy.

Amyloid fibril formation by a normally folded protein in the absence of denaturants and agitation.

The conversion of normally folded proteins into amyloid-like fibrils is an important process in protein chemistry, biology, pathology and biotechnology. This process generally requires harsh conditions, such as pH extremes, organic cosolvents, high temperatures, high pressures or shear forces. Such conditions promote aggregation because they partially unfold structured proteins or allow the sampling of locally unfolded native-like states, both of which possibly represent amyloidogenic states. Here we report the formation of amyloid-like fibrils by the lipase from Pseudomonas sp. under conditions that are close to physiological, that is, in the absence of denaturants and agitation. The resulting aggregates bind thioflavin T and Congo red, causing their characteristic spectral changes observed in the presence of amyloid fibrils. They possess a significant quantity of ß-sheet structure, as detected with Fourier transform infrared and far-UV circular dichroism spectroscopies, and appear fibrillar using transmission electron microscopy. These results indicate that the lipase from Pseudomonas sp. can be a useful model system for the characterization of a key process, such as amyloid fibril formation under physiological conditions.