IMMUNOLOGICAL DISORDERS AND COLON DYSBIOSIS IN OBESE PATIENTS WITH HYPOTHYROIDISM.

OBJECTIVE: The aim: To investigate the peculiarities of immunological changes and their relationship with colon dysbiosis in obese patients with HT. PATIENTS AND METHODS: Materials and methods: The examined patients included 48 patients with HT and obesity (group 1) and 34 patients with obesity (group 2). Patients under¬went fecal analysis for dysbiosis. The levels of complement, namely C3 and C4 and the concentration of immunoglobulins (IgA, Ig M, IgG) were determined by means of chromogenic analysis. RESULTS: Results: During the clinical examination, constipation and flatulence were more often diagnosed in patients of group I (58.3% and 66.7%, respectively - p<0.001), while in patients of group 2 with increased BMI without thyroid dysfunction, a tendency to diarrhea was more often found, accompanied by periodic pain along the colon (50.0% and 32.3% of patients, respectively - p<0.001). Changes in the immunological status of patients in both groups were found. In patients with HT and increase of BMI an increase in serum IgA, IgM, IgG levels were found. An increase in serum immunoglobulins (A, M and G) was also diagnosed in group 2 of examined patients too. CONCLUSION: Conclusions: 1. In patients with obesity decrease in the concentration of Bifidobacterium, Lactobacillus and increase in the number of Staphylococcus, Clostridium, Proteus and Klebsiella were detected, which is more pronounced in patients with a combination of obesity and hypothyroidism. 2. Impairment distinct of immu¬nological status in patients with hypothyroidism and obesity was diagnosed, which was manifested by increased levels of immunoglobulins, namly (A, M, G), as well as a decrease in blood serum complements (C3, C4). 3. The level of IgA, G directly depends on the decrese of Bifidobacterium, Lactobacillus and increse of Staphylococcus, Clostridium and Klebsiella in patients with obesity, which is more pronounced in patients with a combination of obesity and hypothyroidism.

Performance of a commercial industrial-scale UF-based process for treatment of oily wastewaters.

An evaluation was made of the performance of a commercial industrial-scale ultrafiltration (UF)-based process for treatment of highly concentrated oily wastewaters. Wastewater samples were gathered from two plants treating industrial wastewaters in 2008, and in 2011 (only from one of the plants), from three points of a UF-based treatment train. The wastewater samples were analyzed by measuring the BOD7, COD, TOC and total surface charge (TSC). The inorganic content and zeta potentials of the samples were analyzed and GC-FID/MS analyses were performed. The removal performances of BOD7, COD, TOC and TSC in 2008 and 2011 for both plants were very high. Initial concentrations of contaminants in 2011 were lower than in 2008, therefore the COD and TSC reductions were also lower in 2011 than three years before. Regardless of the high performance of UF-based processes in both plants, at times the residual concentrations were considerable. This could be explained by the high initial concentrations and also by the presence of the dissolved compounds that were characterized. Linear correlation was observed between COD and TOC, and between COD and TSC. The correlation between COD and TSC could be utilized for process control purposes.

The effect of beta-glucan on the glycemic and insulin index.

OBJECTIVE: To determine the effects of oat products with increasing beta-glucan content on the glycemic (GI) and insulin indexes (II) of oat products, and to establish the effect of physical properties of beta-glucan on these physiological responses. DESIGN: Test group (n=10) randomly attended to three glucose tolerance tests and glycemic response tests for four oat bran products. SETTINGS: Functional Foods Forum and the Department of Food Chemistry, University of Turku, and the Department of Food Technology, University of Helsinki. SUBJECTS: One male and nine female volunteers were recruited from university students and staff, and all completed the study. INTERVENTIONS: GI and II of different products were calculated for each subject using the average of parallel glucose tolerance tests and the subsequent glycemic/insulinemic responses for each product. Average indexes for products were calculated according to the individual data. RESULTS: The glycemic responses to oat products with increasing amounts of beta-glucan had lower peak values than the reference glucose load. The amount of extractable beta-glucan had a high correlation between the glycemic and insulinemic response. CONCLUSION: In addition to the total amount of beta-glucan in oat products, the amount of extractable beta-glucan in oat products explains the magnitude of the decrease in glycemic responses to carbohydrate products.

Glycemic responses of oat bran products in type 2 diabetic patients.

BACKGROUND AND AIM: Cereal products with low postprandial glycemic response are encouraged in the management of hyperglycemia. In this study, we determined the postprandial glycemic response of two different oat bran products in patients with type 2 diabetes. In addition, we investigated the effects of oat bran flour on postprandial glucose response following an oral glucose load. METHODS AND RESULTS: A randomized, controlled, repeated measures design with two test series was used. Twelve type 2 diabetic patients participated in five 2-h meal glucose tolerance tests on separate occasions. Volunteers were given in random order oat bran flour, oat bran crisp and glucose load providing 12.5 g glycemic carbohydrate (series 1), 25 g glucose load alone and 25 g glucose load with 30 g oat bran flour (series 2). Finger-prick capillary blood analysis was carried out fasting and then 15, 30, 45, 60, 90 and 120 min after the start of the meal. The oat bran flour had a lower 0-120 min area under the glucose response curve (AUC) (47+/-45 mmol min/L) than the glucose load (118+/-40 mmol min/L) (p<0.002), but there was no difference between the oat bran crisp (93+/-41 mmol min/L) and the glucose load in this respect. The oat bran flour decreased the glucose excursion from baseline by 1.6 mmol/l (2.4, 0.8) (mean (95% CI)) and 1.5 mmol/l (2.0, 1.1) at 30 and 45 min after the glucose load, respectively. CONCLUSIONS: Oat bran flour high in beta-glucan had a low glycemic response and acted as an active ingredient decreasing postprandial glycemic response of an oral glucose load in subjects with type 2 diabetes.

Gonadal tumors of mice double transgenic for inhibin-alpha promoter-driven simian virus 40 T-antigen and herpes simplex virus thymidine kinase are sensitive to ganciclovir treatment.

We have previously produced transgenic (TG) mice expressing the mouse inhibin alpha-subunit promoter/Simian virus 40 T-antigen (Inhalpha/Tag) fusion gene. The mice develop gonadal somatic cell tumors at the age of 5-7 months; the ovarian tumors originate from granulosa cells, and those of the testes from Leydig cells. In the present study another TG mouse line was produced, expressing under the same inh-alpha promoter the herpes simplex virus thymidine kinase gene (Inhalpha/TK). Crossbreeding of the two TG mouse lines resulted in double TG mice (Inhalpha/TK-Inhalpha/Tag), which also developed gonadal tumors. The single (Inhalpha/Tag) and double TG (Inhalpha/TK-Inhalpha/Tag) mice, both bearing gonadal tumors, were treated at the age of 5.5-6.5 months with ganciclovir (GCV, 150 mg/kg body weight twice daily i.p.) for 14 days, or with aciclovir (ACV, 300-400 mg/kg body weight per day perorally) for 2 months. During GCV treatment, the total gonadal volume including the tumor, decreased in double TG mice by an average of 40% (P<0.05), while in single TG mice, there was a concomitant increase of 60% in gonadal size (P<0.05). GCV was also found to increase apoptosis in gonads of the double TG mice. Peroral treatment with ACV was less effective, it did not reduce significantly the gonadal volume. We also analyzed the in vitro efficacy of ACV and GCV treatments in transiently HSV-TK-transfected KK-1 murine granulosa tumor cells, originating from a single-positive Inhalpha/Tag mouse. GCV proved to be more effective and more specific than ACV in action. These results prove the principle that targeted expression of the HSV-TK gene in gonadal somatic cell tumors is potentially useful for tumor ablation by antiherpes treatment. The findings provide a lead for further development of somatic gene therapy for gonadal tumors.

Obstetric outcome in women with polycystic ovarian syndrome.

Women with polycystic ovarian syndrome (PCOS) often have insulin resistance and hyperinsulinaemia and may therefore be at an increased risk for gestational diabetes mellitus (GDM). Hyperinsulinaemia may also be associated with pre-eclampsia. Information concerning outcome of pregnancies in PCOS women is scanty and somewhat controversial. Therefore, 99 pregnancies were retrospectively evaluated in women with PCOS and the findings were compared with an unselected control population. The average body mass index (BMI) in PCOS patients was greater than that in controls (25.6 versus 23.0) (P < 0.0001), and PCOS patients were more often nulliparous than controls (76 versus 42%) (P < 0.001). The multiple pregnancy rate was 9.1% in PCOS patients and 1.1% in controls [odds ratio (OR) 9.0; 95% confidence interval (CI) 3.5-23.3]. GDM developed in 20% of the PCOS patients and in 8.9% of the controls (P < 0.001). After logistic regression analysis, BMI >25 seemed to be the greatest predictor for GDM (adjusted OR 5.1; CI 3.2-8.3), while PCOS remained as another independent predictor (adjusted OR 1.9; CI 1.0-3.5). In contrast, PCOS was not a significant predictor for pre-eclampsia, which was merely associated with nulliparity. Premature delivery (16.1% in PCOS and 6.5% in controls) was explained to a large extent by multiple pregnancies and marginally by nulliparity and PCOS. In singleton pregnancies, there was no difference in birth weights, Apgar scores or perinatal morbidity of infants. In conclusion, PCOS slightly increases the risk for GDM, but does not have an important effect on the rate of premature delivery and pre-eclampsia.

The kinetics of substitution reactions of mixed platinum(II)-bis(nucleoside) complexes in aqueous solution in the presence of thiourea; X-ray crystal structure determination of cis-

In aqueous solution, bis(nucleoside) complexes formed by the reaction of cis-[Pt(NH3)2(H2O)2]2+ with an excess of either adenosine (ado) or a mixture of adenosine and guanosine (guo) undergo a slow N7--> N1 linkage isomerisation in the adenine moiety. The isomerisation probably involves the breaking and reformation of Pt-nucleoside bonds, thus favouring the more stable N1 binding mode of the adenine base. Dynamic processes due to the presence of adenosine in the platinum coordination sphere are slow on the NMR time scale. The N7 binding mode of PtII in cis-[Pt(NH3)2(ado-N7)2](ClO4)2. 3.5H2O was confirmed by X-ray crystal structure analysis. In both of the crystallographically independent cations, the PtII coordination sphere is almost ideally square planar, with typical Pt-N bond lengths and angles. The most significant difference between the two cations lies in the sugar conformation of the coordinated nucleosides. In one cation, both have an anti (-ap) conformation, whilst in the other cation one has an anti (-ap) conformation and the other a syn (+sc) conformation stabilised by a relatively strong H-bond. Substitution of the nucleoside(s) by thiourea follows an associative mechanism with only a negligible contribution by the solvent path. For symmetric complexes, the order of lability of different binding modes is ado-N1

Fed-batch fermentor synthesis of 3-dehydroshikimic acid using recombinant Escherichia coli.

3-Dehydroshikimic acid (DHS), in addition to being a potent antioxidant, is the key hydroaromatic intermediate in the biocatalytic conversion of glucose into aromatic bioproducts and a variety of industrial chemicals. Microbial synthesis of DHS, like other intermediates in the common pathway of aromatic amino acid biosynthesis, has previously been examined only under shake flask conditions. In this account, synthesis of DHS using recombinant Escherichia coli constructs is examined in a fed-batch fermentor where glucose availability, oxygenation levels, and solution pH are controlled. DHS yields and titers are also determined by the activity of 3-deoxy-D-arabino-heptulosonic acid 7-phosphate (DAHP) synthase. This enzyme's expression levels, sensitivity to feedback inhibition, and the availability of its substrates, phosphoenolpyruvate (PEP) and D-erythrose 4-phosphate (E4P), dictate its in vivo activity. By combining fed-batch fermentor control with amplified expression of a feedback-insensitive isozyme of DAHP synthase and amplified expression of transketolase, DHS titers of 69 g/L were synthesized in 30% yield (mol/mol) from D-glucose. Significant concentrations of 3-dehydroquinic acid (6.8 g/L) and gallic acid (6.6 g/L) were synthesized in addition to DHS. The pronounced impact of transketolase overexpression, which increases E4P availability, on DHS titers and yields indicates that PEP availability is not a limiting factor under the fed-batch fermentor conditions employed.

In situ mutagenesis and chemotactic selection of microorganisms in a diffusion gradient chamber.

A new method has been developed to rapidly generate and select microbial strains having increased resistance to an inhibitory compound. The method combines in situ mutagenesis with use of a continuous gradient of the inhibitor to sort cells according to their resistance levels. Microbial chemotaxis is induced to accelerate the selection process. The method was used to develop a strain of E. coli having a feedback-resistant DAHP synthase enzyme. An unsteady-state mathematical model of the process has been developed. The model, that can reproduce key trends observed experimentally, was used to explore the effects of chemotaxis on the efficiency of the selection process.

Transgenic mouse models for gonadal tumorigenesis.

The versatile transgenic (TG) techniques allow the production of in vivo animal models for a variety of diseases, including malignant tumors, through tissue-specific expression of oncogenes. We have created a TG mouse model for gonadal somatic cell tumors by expressing the powerful viral oncogene, Simian virus 40 T-antigen (Tag) under regulation of the murine inhibin alpha-subunit promoter (inh alpha). Ovarian granulosa and theca cell tumors were formed in the female, and those of testicular Leydig cells, in the male TG mice at the age of 5-6 months, with 100% penetrance. The tumors produced high levels of inhibin peptides, especially the alpha-subunit, and were steroidogenically active, mainly producing progesterone. The gonadal tumorigenesis was gonadotropin-dependent, since TG mice rendered gonadotropin-deficient by crossbreeding them into the hypogonadotropic hpg genetic background, or by treating them with a gonadotropin-releasing hormone (GnRH) antagonist, did not develop tumors. In order to study the possibility of using the tumor mouse model for testing gene therapy, we created another TG mouse model expressing under the same inhibin-alpha promoter the Herpes Simplex virus (HSV) thymidine kinase (TK) transgene. The inh alpha/HSV-TK mice were crossbred with the inh alpha/Tag mice and the double mutant mice also developed gonadal tumors. When they were treated with antiherpes drugs (acyclovir or gancyclovir), further growth of the tumors was blocked. These preliminary findings prove the principle that tumor ablation in our TG mouse model can be achieved by transduction of the HSV-TK gene into the tumor cells. Besides studies of formation, regulation and therapy of the tumors in vivo, immortalized cell lines derived from them provide models for studies of gonadal somatic cell functions in vitro.

Gonadectomy permits adrenocortical tumorigenesis in mice transgenic for the mouse inhibin alpha-subunit promoter/simian virus 40 T-antigen fusion gene: evidence for negative autoregulation of the inhibin alpha-subunit gene.

We have developed a transgenic (TG) mouse model for gonadal tumorigenesis expressing the Simian virus 40 T-antigen (Tag) under the mouse inhibin alpha-subunit promoter. Gonadal tumors appear with 100% penetrance by the age of 5-8 months in the TG mice. When 1-month-old TG mice were gonadectomized, adrenal gland tumors were observed in each animal (12 females, 11 males) at the age of 6-8 months. No adrenal tumors were detected in gonadectomized non-TG mice (nine females, nine males) or in the intact TG mice (n > 100). The tumors appeared to originate from the X zone of the adrenal cortex. The TG mice with adrenocortical tumors had elevated serum levels of progesterone, estradiol, and immunoreactive inhibin (including dimeric forms), but corticosterone secretion was reduced. The lack of adrenal tumors in intact TG mice suggested that the tumorous gonads secrete factor(s) inhibiting adrenal tumorigenesis. As a candidate molecule, we studied the effects of inhibin, which was high in the serum of control females and TG females with ovarian tumors, as well as in TG males with testicular tumors. The DNA synthesis, as well as the levels of inhibin-alpha and Tag mRNA expression, were significantly reduced by recombinant human inhibin A in cell cultures derived from the adrenal tumors. In accordance, the expression level of inhibin-alpha mRNA in the normal adrenal gland was elevated 2 weeks after gonadectomy. These findings suggest that gonadal inhibin can down-regulate the expression of the inhibin alpha-subunit gene in the adrenal gland. When circulating inhibin is eliminated by gonadectomy, Tag expression and tumorigenesis are stimulated in the adrenal glands of the TG mice. The results demonstrate a novel mechanism of autoregulation in inhibin alpha-subunit gene expression.

Detection of a point mutation using short oligonucleotide probes in allele-specific hybridization.

Two nonradioactive and simple procedures were developed to detect the A985G point mutation that causes medium-chain acyl-CoA deficiency. In both of these assays, short oligonucleotide probes were used in allele-specific hybridization combined with DNA amplification. The lower limit for a useful probe was found to be between 9 and 12 base pairs. Time-resolved fluorometry was utilized as the label technology and microtitration plates as the solid support. In one of the assay formats, probes labeled with europium and samarium chelates were used to simultaneously detect the mutant and normal alleles from the same hybridization reaction. In addition, the discrimination efficiency of different probes was characterized by cross-reactivity determinations and by measuring affinities of the probes towards fully complementary as well as towards mismatch-forming target oligonucleotides. All of the 80 coded patient samples analyzed were correctly typed in both of the assay formats used.