Periodontal and cardiovascular therapies modify specialized pro-resolving lipid mediator (sPRLM) (LPXA4, PD1, RvE1, RvD1, and MaR1)-mediated pathway: the first pilot clinical study.

OBJECTIVES: Cardiovascular disease (CVD) and periodontal disease have a common pathogenesis with inflammation and resolution steps. Although the relationships among periodontal disease, CVD, and specialized pro-resolving lipid mediator (sPRLM)s are well known, there is no study about the combined effects of cardiovascular and periodontal treatments on sPRLM levels. It was aimed to evaluate the effects of periodontal and cardiovascular therapies on sPRLMs (lipoxin A4, protectin (PD)1, resolvin (Rv) E1, RvD1, and maresin (MaR)1) in patients with CVD and periodontal disease. METHODS: This observational study consisted of fifty-five patients with CVD and mild or moderate periodontitis. The clinical periodontal parameters (plaque index, gingival index, probing pocket depth, percentage of bleeding on probing, and clinical attachment level) and blood and unstimulated total saliva samples were obtained at baseline, at 3 months (following only cardiovascular therapy), and at 6 months (following cardiovascular and periodontal therapies). The blood count and serum levels of cardiometabolic biomarkers (white blood cell, neutrophil/lymphocyte, serum total cholesterol (TC), triglyceride, and low and high-density lipoprotein (HDL) levels) were evaluated. sPRLMs were evaluated by ELISA. RESULTS: There were significant decreases in body mass index, clinical periodontal parameters, WBC, LDL, PD1, and RvD1 at 6 months compared to baseline. The decreases in TC/HDL, RvE1, and MaR1 levels were significant at 3 and 6 months compared to baseline (p < 0.05). CONCLUSION(S): The combination of cardiovascular and periodontal treatments leads to significant reductions in clinical periodontal and cardiometabolic parameters and sPRLMs. CLINICAL RELEVANCE: Our report, which is the first in their field, suggested that cardiovascular and periodontal therapies provide an important contribution via decreasing the periodontal and atherosclerotic inflammation modulating sPRLMs. This finding will be a big step toward increasing the quality of life in these patients by drawing attention to importance of public health associated with oral hygiene, periodontal health, and systemic phase of periodontal treatment.

Immunochemical identification of the mammalian peptide hormone obestatin from tea plant (Camellia sinensis).

This study determines obestatin-like substances from the young shoots of the tea plant [Camellia sinensis (L.) O. Kuntze (Theaceae)]. Proteins were extracted from the vegetative tea leaves using the QB (Quick Buffer) buffer as an extraction buffer. Obestatin-like substances in tea extract were investigated using an indirect home-made enzyme-linked immunosorbent assay (ELISA). Human obestatin-like immunoreactive substances from tea extract were isolated and characterized by tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (tricine-SDS-PAGE) and immunoblotting techniques. Immunochemical results showed that there are strong human obestatin-like immunoreactive substances (0.048±0.0064ng/mg protein) in vegetative tea leaves. This finding was completely unexpected since this hormone was considered to be present solely in animals. Furthermore, a single obestatin-like immunoreactive protein band of 13kDa was identified by tricine-SDS-PAGE and Western blotting of extract of vegetative tea leaf proteins. Present investigation is the first report of presence of obestatin-like immunoreactive substances in plants. It is concluded that obestatin-like bioactive peptides derived from plants can affect gastrointestinal tract structures as endogenous obestatin does and hence play a role in appetite regulation and body weight gain.

Synergistic Effect of Omega-3 and Probiotic Supplementation on Preventing Ligature-Induced Periodontitis.

Omega-3 and probiotics were shown to improve periodontal health by modulating the host immune response. Recently, the combination of omega-3 and probiotics has been shown to have a potential synergistic effect on host modulation. The aim of this study was to evaluate the prophylactic role of an omega-3 and probiotic combination on alveolar bone loss (ABL) via inflammatory response in an experimental periodontitis model. Forty-three rats were divided into 5 groups as control (C, n = 8), periodontitis (P, n = 8), omega-3 + periodontitis (O, n = 8), probiotic + periodontitis (Pro, n = 10), and omega-3 + probiotic + periodontitis (OPro, n = 9). Additionally to a standardized diet, omega-3 and/or probiotics were supplemented with oral gavage to the O, Pro, and OPro groups for 44 days. Periodontitis was induced by ligature to the P, O, Pro, and OPro groups on the 30th day for 2 weeks. ABL levels were measured histopathologically, and serum interleukin (IL) 1ß, IL6, and IL10 levels were analysed by enzyme-linked immunosorbent assay. ABL increased in all periodontitis groups (P, O, Pro, and OPro), compared to C group. Compared to P group, all oral gavage groups (O, Pro, and OPro) revealed decreased ABL, which was lowest in OPro group. IL1ß and IL6 decreased and IL10 increased in OPro group, compared to P group. In conclusion, prophylactic administration of omega-3 and probiotic combination reduced ABL and improved serum IL1ß, IL6, and IL10 levels more than their single use.

Circulating levels of ghrelin, galanin, and orexin-A orexigenic neuropeptides in obstructive sleep apnea syndrome.

PURPOSE: The orexigenic peptides, ghrelin, galanin, and orexin-A, have an important role in food intake and energy homeostasis and regulate the higher brain functions including the sleep-wake state. Although the interactions of these neuropeptides affect neuroendocrine systems resulting in obesity, a major risk factor for obstructive sleep apnea syndrome (OSAS), the mechanism has not been fully elucidated. The objective of this study was to evaluate the association of serum ghrelin, galanin, and orexin-A levels with OSAS. METHODS: In this cross-sectional study, patients who underwent one-night polysomnography and conformed to the inclusion criteria were asked to participate. A blood sample was obtained from all participants on the morning of the sleep test to evaluate the serum levels of ghrelin, galanin, and orexin-A using the enzyme-linked immunosorbent assay (ELISA) method. Demographic characteristics, polysomnography data, and serum levels of the participants were recorded and analyzed. Comparison between the OSAS groups was performed by independent sample t-test, Mann-Whitney U test, and Kruskal-Wallis test with post hoc K-W test using SPSS 20.0. RESULTS: Of 272 patients, those in the OSAS group (n=210) were older than patients in the non-OSAS group (n=62), p < 0.003, and had increased BMI, p < 0.006. Patients with, serum ghrelin, galanin, and orexin-A levels were significantly elevated in patients with OSAS (635.9 pg/mL vs. 420.7 pg/mL, 91.0 pg/mL vs. 60.0 pg/mL, 600.3 pg/mL vs. 485.6 pg/mL, respectively) and found to be higher in patients with severe OSAS than mild and moderate cases (p < 0.01). In multinomial logistic regression to predict the OSAS severity, levels of serum ghrelin (OR = 1.016 [1.010-1.021]; p < 0.001), galanin (OR = 1.050 [1.020-1.081]; p < 0.001), and orexin-A (OR = 1.021 [1.012-1.030]; p < 0.001) were significantly associated only with a moderate level of OSAS. CONCLUSION: The orexigenic neuropeptides were found to be an independent determinant of the presence of OSAS and correlate with the severity of OSAS. Increased levels of ghrelin, galanin, and orexin-A were associated with the presence of moderate OSAS.

Protective Effects of Quercetin on Oxidative Stress-Induced Tubular Epithelial Damage in the Experimental Rat Hyperoxaluria Model.

Background and Objectives: The most common kidney stones are calcium stones and calcium oxalate (CaOx) stones are the most common type of calcium stones. Hyperoxaluria is an essential risk factor for the formation of these stones. Quercetin is a polyphenol with antioxidant, anti-inflammatory, and many other physiological effects. The aim of this study was to investigate the protective effect of quercetin in hyperoxaluria-induced nephrolithiasis. Materials and Methods: Male Wistar-Albino rats weighing 250-300 g (n = 24) were randomized into three groups: Control (n = 8), ethylene glycol (EG) (n = 8), and EG + quercetin (n = 8). One percent EG-water solution was given to all rats except for the control group as drinking water for five weeks. Quercetin-water solution was given to the EG + quercetin group by oral gavage at a dose of 10 mg/kg/day. Malondialdehyde (MDA), catalase (CAT), urea, calcium, and oxalate levels were analyzed in blood and urine samples. Histopathological assessments and immunohistochemical analyses for oxidative stress and inflammation indicators p38 mitogen-activated protein kinase (p38-MAPK) and nuclear factor kappa B (NF-kB) were performed on renal tissues. Results: The MDA levels were significantly lower in the quercetin-treated group than in the EG-treated group (p = 0.001). Although CAT levels were higher in the quercetin-treated group than the EG-administered group, they were not significantly different between these groups. The expression of p38 MAPK was significantly less in the quercetin-treated group than the EG group (p < 0.004). There was no statistically significant difference between the quercetin and EG groups in terms of NF-kB expression. Conclusions: We conclude that hyperoxaluria activated the signaling pathways, which facilitate the oxidative processes leading to oxalate stone formation in the kidneys. Our findings indicated that quercetin reduced damage due to hyperoxaluria. These results imply that quercetin can be considered a therapeutic agent for decreasing oxalate stone formation, especially in patients with recurrent stones due to hyperoxaluria.

Salivary levels of last generation specific pro-resolving lipid mediators (SPMs) (protectin and maresin) in patients with cardiovascular and periodontal disease: A case-control study.

BACKGROUND AND OBJECTIVE: Periodontal disease and cardiovascular disease (CVD), which are both deemed to be triggered by inflammation, are recognized as public health problems. Evidence of host modulation via pro-resolving lipid shown in previous studies supports a two-way relationship between periodontitis and CVD. Last generation endogenous specific pro-resolution lipid mediators (SPMs) such as protectins (PDs) and maresins (MaRs) may have potential effects on inflammatory pathogenesis via activation and resolution mechanisms. Currently, there are no data on SPM levels in patients with CVD and periodontal disease. We aimed to evaluate salivary levels of PD and MaR in patients with CVD and periodontal disease. MATERIALS AND METHODS: At total of 181 individuals comprising of 79 healthy controls (C) and 102 patients with diagnosed CVD were included cross-sectionally. Unstimulated total salivary samples were obtained, and clinical periodontal parameters were determined. Salivary levels of PD and MaR were evaluated by ELISA. The periodontal status of the study population was classified as gingivitis (g) or periodontitis (p). RESULTS: Patients with CVD showed lower sociodemographic characteristics, increased clinical periodontal parameters (p < .05), decreased salivary PD (p < .001), and increased salivary MaR levels (p > .05). In the CVDg group, leukocyte, hemoglobin, hematocrit, and high-density lipoprotein values were higher (p < .05). The CVDp group had a higher neutrophil-to-lymphocyte ratio (p < .05). While the PD level was highest in the Cg group, MaR was highest in the CVDp group. The salivary levels of PD and MaR were independent of other confounders in CVD and periodontal disease (p > .05). CONCLUSION(S): PDs and MaRs may play effective roles in pathogenesis associated with worsening cardiometabolic and periodontal status. These SPMs could also be predictors for conversion from a healthy (systemically and periodontally) to diseased state (CVD and/or periodontitis). Elucidation of the role of SPMs in the relationship between periodontal disease and CVD will enable the development of new host modulation strategies in the prevention and treatment of both diseases, and may also constitute an important public health step by increasing the quality of life of patients with CVD and periodontal disease.

Does metabolic control affect salivary adipokines in type 2 diabetes mellitus?

BACKGROUND: Adipokines produced by adipose tissue initiate pro-inflammatory events and contribute to the pathogenesis of diabetic periodontitis. OBJECTIVES: The aim of this study was to evaluate the effect of the metabolic status on the level of salivary adipokines in type 2 diabetes mellitus (T2DM) patients. MATERIAL AND METHODS: A total of 239 individuals, including 161 T2DM patients and 78 healthy (H) controls, participated in the study. The metabolic control status was evaluated in each person. Periodontal measurements were recorded. Periodontal epithelial surface area (PESA), periodontal inflamed surface area (PISA) and the total dental index (TDI) were calculated. The salivary adiponectin, tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and vaspin levels were determined. RESULTS: The T2DM patients had higher periodontal parameters and adiponectin, TNF-α, IL-6, and vaspin levels as compared with the H controls (p < 0.05). As the metabolic control worsened, periodontal pocket depth (PPD) and clinical attachment level (CAL) increased. When covariates (age, gender, body mass index - BMI, education level, smoking, dental visit and tooth brushing frequency) were adjusted, only the TNF-α and vaspin levels were significantly higher in the T2DM patients (p < 0.05). In the T2DM patients, positive correlations were found between the TNF-α level and the percentage of bleeding on probing (BOP%), PPD, PESA, and PISA, and between the adiponectin level and PISA. Moreover, there was a negative relationship between the salivary volume and TDI. While the correlations IL-6-TNF-α, vaspin-triglycerides and vaspin-tooth brushing frequency were positive, the statistically significant associations vaspin-IL-6 and vaspin-low-density lipoprotein (LDL) were negative (p < 0.05). CONCLUSIONS: The severity of periodontal disease increases as the metabolic control status worsens. The levels of salivary adipokines were changed by T2DM, while being independent from the metabolic control.

Determination of Apoptotic Effect of Juglone on Human Bladder Cancer TCC-SUP and RT-4 Cells: An In Vitro Study.

This study aimed to investigate the effects of juglone on the human bladder carcinoma cell lines TCC-SUB and RT-4 in monolayer and spheroid cultures. Cells were treated with juglone at 24, 48, and 72 h of incubation. The activity of caspase-3 was detected in vitro using a caspase-3 colorimetric assay kit according to the manufacturer's instructions. The bromodeoxyuridine (BrdU) labeling index was used to determine the cells of the synthesis phase. The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay was used to determine the death of cells in both the monolayer and spheroid cultures. The control group had a large S-phase fraction and many of the TCC-SUB and RT-4 cells nuclei were observed to be positive for BrdU. The dead cell count was higher in the TCC-SUB and RT-4 cell lines with juglone applied than in the controls. We conclude that juglone significantly inhibits the proliferation and induces the apoptosis of TCC-SUB and RT-4 cells in vitro.

Protective impact of resveratrol in experimental rat model of hyperoxaluria.

PURPOSE: Resveratrol (RES) is a polyphenol with antioxidant, anti-inflammatory, and many other physiological effects on tissues. In the present study, the effect of resveratrol in hyperoxaluria driven nephrolithiasis/nephrocalcinosis is investigated. METHODS: Wistar-Albino rats of 250-300 g (male, n = 24) were included in the present study. The rats were randomized into three groups: Group 1 consisted of the controls (n = 8), Group 2 of hyperoxaluria (1% ethylene glycol (EG), n = 8), and Group 3 of the treatment (1% EG + 10 mg/kg of RES, n = 8) group. At the beginning and fifth week of the study, two rats from each group were placed in metabolic cages for 24 h and their urine was collected. At the end of the study, the rats were killed and their blood was collected from the vena cava inferior. The right kidneys of the rats were used for biochemical and the left ones for immunohistochemical analyzes. Malondialdehyde (MDA), catalase, urea, calcium, oxalate, and creatinine clearance were studied in the blood, urine, and kidney tissues. Moreover, routine histological evaluation, and p38-MAPK and NFkB immunohistochemical analyses were conducted. RESULTS: In the hyperoxaluria group, urinary oxalate levels were higher than the control group; yet, lower in the treatment group compared to hyperoxaluria group (p < 0.05). Serum MDA levels in the hyperoxaluria group were higher than the control group; but in the treatment group it is lower than the hyperoxaluria group (p < 0.05). P38 MAPK activity was higher in the hyperoxaluria group compared to the control (p < 0.05). However, in terms of p38 MAPK activity, there were no statistically significant difference between hyperoxaluria and the treatment group (p < 0.069). Whereas NFkB activity in the hyperoxaluria group is higher than the control (p < 0.001), no statistically significant difference was observed with the treatment group. CONCLUSIONS: In the present study, resveratrol was seen to prevent hyperoxaluria. With preventing oxidative stress factors and Randall plaque formation caused by free oxygen radicals, resveratrol can be an alternative treatment option that can increase the success rate in preventing stone recurrence in the future.

A Case-Control Association Study of RANTES (-28C>G) Polymorphism as a Risk Factor for Parkinson's Disease in Isparta, Turkey.

Background. Recent studies have revealed that inflammatory processes are involved in the pathogenesis of Parkinson's disease (PD). Multiple lines of evidence have suggested that chemokines and their receptors are involved in several neurodegenerative disorders. We have examined whether genetic polymorphisms at the genes encoding chemokines IL-8 (-251A>T), MCP-1 (-2518A/G), and RANTES (-28C>G) and chemokine receptors CCR2 (V64I) and CCR5 (-Δ32) were associated with sporadic PD risk in Isparta, Turkey. Method. The pilot case-control association study included 30 PD patients and 60 control subjects, who were all genotyped with PCR-RFLP for the five polymorphisms. Their genotype and haplotype frequencies were compared statistically. Results. One SNP (-28C>G) in RANTES revealed a significant association with PD (P (allele) < 0.0001, p-trend = 0.0007). The risk allele (G) in the homozygous and dominant models (OR = 17.29 and 32.10, 95% CI = 0.86-347.24 and 1.74-591.937, resp.) suggests additional PD risk. The haplotype TGCAN from the IL-8 (-251A>T), MCP-1 (-2518A>G), RANTES (-28C>G), CCR-2 (V64I), and CCR-5 (-Δ32) has protective effect (OR = 0.08 [CI = 0.01-0.63], p = 0.019). Conclusions. Our data are the first indication of the role of RANTES (-28C>G) in PD risk.

The effects of atorvastatin therapy on rheological characteristics of erythrocyte membrane, serum lipid profile and oxidative status in patients with dyslipidemia.

The statins, most commonly used in the treatment of hyperlipidemia, have certain beneficial effects including improved endothelial function, plaque stability and decreased oxidative stress and inflammation, beyond their lipid-lowering effect in plasma. We evaluated the pleiotropic impact of atorvastatin on erythrocyte structural/mechanical properties and lipid peroxidation in dyslipidemics. The study group included 44 patients with dyslipidemia and was divided into subgroups according to triglyceride and cholesterol levels as hypercholesterolemic (n = 29) and mixed-type hyperlipidemic (n = 15). Subjects were given 10 mg atorvastatin per day for 12 weeks. Changes in serum lipid composition, lipid contents, Na(+)/K(+)-ATPase activity and osmotic fragility in erythrocytes and oxidative stress parameters of erythrocytes and plasma were studied. Atorvastatin therapy improved the serum lipid profile of both subgroups. This alteration was accompanied by a decreased level of cholesterol in erythrocyte membranes. Moreover, enhanced activity of Na(+)/K(+)-ATPase in erythrocytes reflected the improvements in membrane lipids of both subgroups. However, a significant change was observed in osmotic fragility values of the mixed-typed dyslipidemic group. This treatment lowered the lipid peroxidation in plasma and erythrocytes and increased plasma total antioxidant capacity in all groups. The present study shows that the use of atorvastatin reversed the structural and functional features of erythrocyte membranes in dyslipidemic subjects. Also, hypolipidemic therapy had a beneficial impact on a balance between oxidant and antioxidant systems.

Effects of misoprostol on cisplatin-induced renal damage in rats.

Cisplatin (CP) is a potent anticancer drug. However, it has side effects on kidney such as nephrotoxicity. Abnormal production of reactive oxygen species (ROS) has been accused in the etiology of CP-induced nephrotoxicity. Several ROS scavengers have been reported to prevent nephrotoxicity after CP administration. In this study, we used prostaglandin E1 (PGE1) analogues misoprostol (MP) to reduce this damage. MP has gained considerable interest as a ROS scavenger. Rats were received a single injection of CP (5 mg/kg, i.p.) with or without MP pretreatment (200 mcg/kg, orally). The renal tissue morphology was investigated by light microscopy. Trunk blood was also obtained to determine lipid peroxidation product malondialdehyde (MDA) and activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT). CP administration increased MDA production and decreased SOD and CAT levels in the kidney tissue when compared to the control group. Morphological damage in CP administrated rats was also severe in the kidney tissue. MP treatment after CP application protected the renal tissues from CP's side effect. These findings indicate that MP has beneficial effects on CP induced nephrotoxicity in rats.

Serotonin transporter bi- and triallelic genotypes and their relationship with anxiety and academic performance: a preliminary study.

BACKGROUND: Considerable evidence suggests that variation of the serotonin-transporter-linked promoter region (5- HTTLPR) is associated with anxiety-related traits. Academic outcomes are also more closely related to trait anxiety. This preliminary study aimed to explore the association between academic performance and levels of anxiety with respect to the bi- and triallelic classification of 5-HTTLPR polymorphism of the 5-HTT gene in teacher candidates. METHODS: In our study, Spielberger's State-Trait Anxiety Inventory, the Selection Examination for Professional Posts in Public Organizations (KPSS) and 5-HTTLPR genotypes were used to investigate a group of 94 healthy teacher candidates. RESULTS: Higher anxiety scores were significantly associated with the S'S' genotype. There was no direct, statistically significant association between academic performance and genotypic groups regarding bi- and triallelic classification. However, the students who have L'L' or LL genotypes had the lowest levels of trait anxiety and the poorest academic performance. Additionally, there was a significant positive correlation between academic performance and anxiety levels. DISCUSSION: These findings support the idea that S and L(G) alleles are associated with anxiety-related traits, and that the S'S' genotype may be a good indicator for anxiety-related traits in a sample from the Turkish population. A specific degree of anxiety is considered to be a motivation for learning and high academic performance. However, 5-HTTLPR polymorphism of the 5-HTT gene may be one of the genetic factors affecting academic performance in connection with anxiety levels. Implications for incorporating anxiety management training in the educational process in terms of both environmental and individual factors will have a very important role in improving effective strategies for student personality services, as well as for development and planning.

Genetic study of demyelinating form of autosomal-recessive Charcot-Marie-tooth diseases in a Turkish family.

Charcot-Marie-Tooth (CMT) disease is a clinically and genetically heterogeneous group of inherited peripheral motor and sensory neuropathies characterized by distal muscle weakness atrophy predominantly in the lower extremities, diminished or absent deep tendon reflexes, distal sensory loss and skeletal deformities. Mode of inheritance could be either autosomal dominant, autosomal recessive, or X-linked. The autosomal-recessive subgroup of CMT (AR-CMT) neuropathies is heterogeneous as well. To date, nine demyelinating loci have been implicated in CMT4 and seven genes have been identified. It has been screened in this study for the presence of mutations in the coding region of GDAP1 and genetic linkage analyses of CMT4B1, CMT4B2, CMT4C, CMT4D, CMT4E, and CMT4F loci were tested in a Turkish family presenting recessively inherited form of CMT disease characterized by severe motor weakness. We did not find any mutations in GDAP1 and genetic linkage excluded for the six demyelinating genes loci (CMT4B1, CMT4B2, CMT4C, CMT4D, CMT4E, and CMT4F). Our findings indicate that another locus may be associated with AR-CMT disease.

Novel GDAP1 mutation in a Turkish family with CMT2K (CMT2K with novel GDAP1 mutation).

Mutations in the ganglioside-induced differentiation-associated protein 1 gene (GDAP1) cause Charcot-Marie-Tooth type 2 (CMT2), a severe autosomal recessive form of neuropathy associated with axonal phenotypes. It has been screened in this study for the presence of mutations in the coding region of GDAP1, which maps to chromosome 8q21, in a family with CMT2. To date, 29 mutations in the GDAP1 have been reported in patients of different ethnic origins. Here, we report a novel missense mutation (c.836A>G), and two polymorphisms: a silent variant (c.102G>C), and a 5'-splice site mutation (IVS5+24C>T) in GDPA1 gene identified in a five generation Turkish family with autosomal recessive CMT2.

The novel ischemia marker 'ischemia-modified albumin' is increased in normal pregnancies.

The aim of this prospective case-control study was to determine the change in serum maternal ischemia-modified albumin (IMA) during normal pregnancies. A total of 117 pregnant (first trimester (n=24), second trimester (n=34), and third trimester (n=35)) and non-pregnant healthy women (n=23) were included. Maternal serum IMA, Malondialdehyde (MDA), and albumin levels were measured. Compared with non-pregnant women, the cross-sectional mean IMA levels in pregnant women were significantly increased, while the mean serum MDA and albumin levels were significantly decreased throughout pregnancy. Furthermore, a significant negative correlation between serum IMA and albumin levels (r=-0.354, p<0.001) was found, and there was a weak positive correlation between serum albumin and MDA levels (r=0.334, p<0.001). Serum IMA, which has recently been developed as a clinical marker of ongoing myocardial ischemia, appears to be elevated in normal pregnancy. This may be due to the physiologic oxidative stress state of pregnancy.

Analysis of the alpha-1-antitrypsin deficient alleles M3S, MZ, and ZZ by biochemical and molecular methods: a family study.

Deficiency of alpha-1-antitrypsin (alpha(1)-AT, a major protease inhibitor controlling tissue degradation) is a genetic disorder transmitted in a codominant autosomal form. It has more than 100 genetically determined variants. This study attempted to determine the degree of association between serum alpha(1)-AT levels and phenotypes and to provide a strategy for reliable laboratory evaluation of deficiencies. The study group consisted of a 38-year-old male proband with clinical features of emphysema, his first-degree relatives, and healthy controls. Family history revealed a four-generation pedigree. Genomic DNA was isolated from peripheral blood leukocytes. Alpha-1-AT levels were determined from human serum by immunonephelometry. Phenotypes were determined by isoelectric focusing of blood samples. DNA sequences of coding exons were analyzed by the amplification DNA technique and direct sequencing. Inheritance and plasma levels of the ZZ, MM, M3S, and MZ phenotypes were confirmed by the family study. In the family members with deficiencies, plasma concentrations were 22.55% +/- 5.15 (ZZ), 84.18% +/- 5.18 (M3S), and 61.06% +/- 7.15 (MZ) of the normal MM level. We found a close association between alpha(1)-AT level and genotype. A combination of genotyping, quantification, and phenotyping is the optimal strategy for the laboratory evaluation of alpha(1)-AT deficiency.

Synthesis, acidity and antioxidant properties of some novel 3,4-disubstituted-4,5-dihydro-1H-1,2,4-triazol-5-one derivatives.

3-Alkyl(aryl)-4-amino-4,5-dihydro-1H-1,2,4-triazol-5-ones 2a-g reacted with 4-diethylaminobenzaldehyde to afford the corresponding 3-alkyl(aryl)-4-(4-diethyl-aminobenzylidenamino)-4,5-dihydro-1H-1,2,4-triazol-5-ones 3a-g. The acetylation reactions of compounds 3a-e were investigated and compounds 4a-e were thus obtained. The new compounds were characterized using IR, (1)H-NMR, (13)C-NMR, UV and MS spectral data. In addition, the newly synthesized compounds 3a-g were titrated potentiometrically with tetrabutylammonium hydroxide in four non-aqueous solvents such as isopropyl alcohol, tert-butyl alcohol, acetone and N,N-dimethylformamide (DMF), and the half-neutralization potential values and the corresponding pKa values were determined for all cases. Moreover, 3 and 4 type compounds were also screened for their antioxidant activities.

Preparation, GIAO NMR calculations and acidic properties of some novel 4,5-dihydro-1H-1,2,4-triazol-5-one derivatives with their antioxidant activities.

Six novel 3-alkyl(aryl)-4-(p-nitrobenzoylamino)-4,5-dihydro-1H-1,2,4-triazol-5- ones (2a-f) were synthesized by the reactions of 3-alkyl(aryl)-4-amino-4,5-dihydro-1H- 1,2,4-triazol-5-ones (1a-f) with p-nitrobenzoyl chloride and characterized by elemental analyses and IR, (1)H-NMR, (13)C-NMR and UV spectral data. The newly synthesized compounds 2 were titrated potentiometrically with tetrabutylammonium hydroxide in four non-aqueous solvents such as acetone, isopropyl alcohol, tert-butyl alcohol and N,N-dimethylformamide, and the half-neutralization potential values and the corresponding pK(a) values were determined for all cases. Thus, the effects of solvents and molecular structure upon acidity were investigated. In addition, isotropic (1)H and (13)C nuclear magnetic shielding constants of compounds 2 were obtained by the gauge-including-atomic-orbital (GIAO) method at the B3LYP density functional level. The geometry of each compound has been optimized using the 6-311G basis set. Theoretical values were compared to the experimental data. Furthermore, these new compounds and five recently reported 3-alkyl-4-(2-furoylamino)-4,5-dihydro-1H-1,2,4-triazol-5-ones (3a-c,e,f) were screened for their antioxidant activities.