Structural, functional, and lectin histochemical characteristics of rat ovaries and endometrium in experimental hyper- and hypothyroidism

Twenty lectins with different carbohydrate affinities, including five new lectins purified from fungi,were used for histochemical examination of carbohydrate determinants in rat ovaries and endometrium underexperimental hyper- and hypothyroidism. In the ovaries of control rats, lectin binding was detected predominantlyin the zona pellucida and corona radiata of growing follicles, luteocytes, and leucocytes. Within the endometrium,lectins reacted strongly with the luminal surface, epitheliocytes and secretions of the uterine glands,pre-decidual cells, leucocytes, and collagen fibers. As revealed on general morphology and estrous cycle studies,hyperthyroidism induced ovarian luteinization, and increased the content of pre-decidual cells and of collagenfibers in the endometrium, with no reliable effect on the estrous cycle. Hypothyroidism was accompanied byretardation of the estrous cycle, enhanced folliculogenesis and leucocyte infiltration of ovarian and endometrialstroma. Both hyper- and hypothyroidism significantly affected the tissue carbohydrates of the ovaries and endometrium:specific and differential redistribution of DMan, LFuc, NeuNAc, DGlcNAc, DGalNAc, and DGaldeterminants was detected under both pathological conditions. Hyperthyroidism induced more severe alterationsof glycoconjugates than hypothyroidism; the endometrium was more susceptible to thyroxin-modulatedimpairments than the ovaries. These results give new insights into the pathogenetic mechanisms of the effects ofthyroid disorders on the female reproductive organs and demonstrate the suitability of lectin histochemistrymethods for monitoring the efficacy of hormonal imbalance correction therapy, as well as the applicability ofnew lectin preparations for the selective labeling of ovarian and endometrial constituents.Twenty lectins with different carbohydrate affinities, including five new lectins purified from fungi,were used for histochemical examination of carbohydrate determinants in rat ovaries and endometrium underexperimental hyper- and hypothyroidism. In the ovaries of control rats, lectin binding was detected predominantlyin the zona pellucida and corona radiata of growing follicles, luteocytes, and leucocytes. Within the endometrium,lectins reacted strongly with the luminal surface, epitheliocytes and secretions of the uterine glands,pre-decidual cells, leucocytes, and collagen fibers. As revealed on general morphology and estrous cycle studies,hyperthyroidism induced ovarian luteinization, and increased the content of pre-decidual cells and of collagenfibers in the endometrium, with no reliable effect on the estrous cycle. Hypothyroidism was accompanied byretardation of the estrous cycle, enhanced folliculogenesis and leucocyte infiltration of ovarian and endometrialstroma. Both hyper- and hypothyroidism significantly affected the tissue carbohydrates of the ovaries and endometrium:specific and differential redistribution of DMan, LFuc, NeuNAc, DGlcNAc, DGalNAc, and DGaldeterminants was detected under both pathological conditions. Hyperthyroidism induced more severe alterationsof glycoconjugates than hypothyroidism; the endometrium was more susceptible to thyroxin-modulatedimpairments than the ovaries. These results give new insights into the pathogenetic mechanisms of the effects ofthyroid disorders on the female reproductive organs and demonstrate the suitability of lectin histochemistrymethods for monitoring the efficacy of hormonal imbalance correction therapy, as well as the applicability ofnew lectin preparations for the selective labeling of ovarian and endometrial constituents

Diabetic alteration versus postnatal maturation of rat kidney glycoconjugates — comparative detection by lectin probes

A panel of ten lectins with different carbohydrate specificities, including three original lectin preparations (MPFA, LABA, and LVA), was used for the investigation of rat kidney glycoconjugate remodeling during postnatal morphogenesis, and the findings were compared with the impairments seen in streptozotocin-induced diabetic nephropathy. Postnatal morphogenesis was accompanied by the accumulation and generalization of DMan, LFuc, and NeuNAc, with simultaneous reduction of DGal and DGalNAc sugar determinants and enhanced heterogeneity of renal microstructure. The most significant redistribution of lectin receptor sites was detected between postnatal days 1 and 20. Beginning from postnatal day 20, renal corpuscles showed selective MPFA, WGA, and RCA labeling. Stabilization of carbohydrate determinants on postnatal day 60 coincided with rat kidney maturity. Diabetic nephropathy induced carbohydrate remodeling reciprocal to that seen during postnatal development, that is, enhanced exposure of DGal and DGalNAc, reduced reactivity of DMan, LFuc, and NeuNAc determinants, and increased lectin labeling of renal tubule brush borders. These results extend the existing data on rearrangement of rat kidney glycoconjugates under physiological and pathological conditions, as well as demonstrate the applicability of three original lectin preparations in glycoconjugate histochemistry. (Folia Histochemica et Cytobiologica 2013, Vol. 51, No. 1, 92–102

Visualization of melanoma tumor with lectin-conjugated rare-earth doped fluoride nanocrystals

Aim To develop specific fluorescent markers for melanoma tumor visualization, which would provide high selectivity and reversible binding pattern, by the use of carbohydrate- recognizing proteins, lectins, combined with the physical ability for imaging deep in the living tissues by utilizing red and near infrared fluorescent properties of specific rare-earth doped nanocrystals (NC). Methods B10F16 melanoma cells were inoculated to C57BL/6 mice for inducing experimental melanoma tumor. Tumors were removed and analyzed by lectin-histochemistry using LABA, PFA, PNA, HPA, SNA, GNA, and NPL lectins and stained with hematoxylin and eosin. NPL lectin was conjugated to fluorescent NaGdF4:Eu3+-COOH nanoparticles (5 nm) via zero length cross-linking reaction, and the conjugates were purified from unbound substances and then used for further visualization of histological samples. Fluorescent microscopy was used to visualize NPL-NaGdF4:Eu3+ with the fluorescent emission at 600-720 nm range. Results NPL lectin selectively recognized regions of undifferentiated melanoblasts surrounding neoangiogenic foci inside melanoma tumor, PNA lectin recognized differentiated melanoblasts, and LCA and WGA were bound to tumor stroma regions. NPL-NaGdF4:Eu3+ conjugated NC were efficiently detecting newly formed regions of melanoma tumor, confirmed by fluorescent microscopy in visible and near infrared mode. These conjugates possessed high photostability and were compatible with convenient xylenebased mounting systems and preserved intensive fluorescent signal at samples storage for at least 6 months. Conclusion NPL lectin-NaGdF4:Eu3+ conjugated NC permitted distinct identification of contours of the melanoma tissue on histological sections using red excitation at 590- 610 nm and near infrared emission of 700-720 nm. These data are of potential practical significance for development of glycans-conjugated nanoparticles to be used for in vivo visualization of melanoma tumor

First Irish delivery following sequential, two-stage embryo and blastocyst transfer.

BACKGROUND: The timing of embryo transfer (ET) after in vitro fertilisation (IVF) remains controversial, and there are no reliable guidelines available to prospectively identify which patients would benefit from either day-3 or blastocyst transfer. While blastocyst transfer is generally favoured over day-3 transfers, very few IVF patients get both in the same treatment cycle. CASE DESCRIPTION: We report on a 35.5-year-old female with tubal factor infertility who underwent IVF, which included transfer of a fresh day-3 embryo and a thawed blastocyst frozen at day 6. Transfer occurred on two separate days (days 3 and 6) in a two-stage/dual catheter fashion and resulted in a healthy term singleton livebirth. CONCLUSIONS: While combined day-3 and day-5 ET has been available elsewhere for several years, this is the first description of its successful application in Ireland and confirms the effectiveness of coordinated two-stage transfer in a single IVF treatment cycle

Persistent accumulation of 4-hydroxynonenal-protein adducts in gastric mucosa after Helicobacter pylori eradication [Povećano nakupljanje 4-hidroksinonenal-proteinskih konjugata u želučanoj sluznici zaostaje nakon izlječenja Helicobacter pylori]

Recent studies indicate that oxidative stress caused by Helicobacter pylori and insufficient host antioxidant defense could play important role in pathogenesis of gastrointestinal ulcerations. By specific monoclonal antibodies we have detected weak presence of the major lipid peroxidation bioactive marker 4-hydroxynonenal (HNE) in healthy human gastric mucosa, which strongly increased in case of H. pylori-associated peptic ulcer. Considering physiological presence of HNE on one hand, and high prevalence of H. pylori associated disorders on the other, evaluation of oxidative stress after treatment is important. Therefore, in current study immunohistochemical accumulation and distribution of HNE-protein adducts in gastric mucosa was evaluated with 21 patients having H. pylori-associated duodenal peptic ulcer (DPU) before and one month after eradication of H. pylori. Although dramatic decrease in histological manifestations of inflammation was demonstrated after eradication of H. pylori, initially high immunopositivity for the HNE-protein adducts remained elevated in antrum and even increased in stomach corpus. The observed accumulation and redistribution to higher grades of HNE-immunopositivity in nuclei of glandular cells in gastric corpus indicate augmentation of oxidative stress after treatment and open possibilities for adjuvant antioxidant treatments to protect gastric mucosa from progressive oxidative stress after eradication of H. pylori infection

Application of the Pierotti Approach in Calculation of the Solubility of Nonelectrolytes in Water-Acid Systems

Based on the experimental values on the solubility of noble gases, nitrogen, oxygen, hydrogen, and a number of hydrocarbons, the possibility of application of the Pierotti approach to predict the solubility of nonelectrolytes in water, acetic acid, inorganic acids, and water-acid mixed solvents over the entire range of their composition was analyzed. The parameters of the Lennard-Jones potential for sulfuric and nitric acids are calculated for the first time. This approach makes it possible to quantitatively predict the solubility of a wide number of nonelectrolytes, including C5-C8 hydrocarbons, in sulfuric and nitric acids and to quantitatively calculate the solubility of small molecules and qualitatively calculate that of long-chain hydrocarbons in acetic acid. In the case of aqueous solutions of hydrocarbons, however, the Pierotti approach fails to predict solubility even at a qualitative level

Solubility of Hydrocarbons in Acetic Acid

Equilibrium distribution coefficients a = [RH]gas/[RH]sol were measured for solutions of normal, iso-, and cycloalkanes and benzene in 100% acetic acid (AcOH) at 298 K and for methane in H2O-AcOH over the whole range of mixture compositions at 283-328 K. Nonelectrolyte solutions obey the equations log(SA/SW) = a + bV̄NE and (logS)E = LVE/V relating the solubility of the nonelectrolyte (NE) in the acid (SA) to its partial molar volume (V̄NE) and solubility in water (SW) (the first equation) and excess solubility values (logS)E to the molar volume (VE) of the system (the second equation)

A New Calculation Method for Arenes Solubilities in the Whole Composition Range of Water-Sulfuric Acid System

Rules were developed to calculate the solubility of non-electrolytes including arenes and their derivatives in the water-sulfuric acid system over the entire range of composition. The approach predicts the solubility in the cases when direct measurements are impossible because of difficulties. Two examples were considered of applications of the rules formulated for benzene and nitrobenzene solutions in the sulfuric acid-water system. The solubility of benzene was measured at 0-90% sulfuric acid. The limiting distribution coefficient α of benzene over the entire range of compositions of the water-sulfuric acid system was calculated. The experimental and calculated data at 0-80% sulfuric acid agreed well. Thus, the calculated values for a more concentrated acid should also be correct. The solubility of nitrobenzene in the water-sulfuric acid system was measured by Hammett and Chapman (1934) at 0-79.3% at 298.15 K. The α values were calculated for this system. The calculated solubility was lower than the experimental one because nitrobenzene interacts with the water-sulfuric acid system