Histochemical demonstration of mucosubstances in the mouse gastrointestinal tract treated with Origanum hypericifolium O. Schwartz and P.H. Davis extract

Glycoconjugate-containing mucous in the gastrointestinal tract has lubricant and cytoprotective effects. This study was performed to demonstrate the intensity of mucous in the stomach, small and large intestine tissues of female BALB/c mice injected intraperitoneally with 0.2 ml of the Origanum hypericifolium extract for six weeks. The staining procedures employed were Alcian Blue (AB) at pH 1 and 2.5, periodic acid-Schiff (PAS) and lectins including Galanthus nivalis agglutinin (GNA), Arachis hypogaea (PNA, Peanut Agglutinin) and Maackia amurensis leucoagglutinin (MAL). The mucous glands in the large intestine of the experimental groups were found to show stronger positive reaction for AB at pH 2.5 and PAS when compared with the controls. In lectin histochemistry, the mucous glands which reacted with lectins exhibited less density for PNA in the stomach and for MAL and PNA in the small intestine in the experimental groups than the controls. In the large intestine of the experimental groups, less staining pattern of GNA was observed on the surface epithelium of villi when compared with the controls. The results suggest that O. hypericifolium extract increases acidic and neutral mucosubstances in the large intestine and also changes some glycan structures in the gastrointestinal tract

The investigation of the effects of Crataegus aronia var. dentata Browicz extract on spleen: a histochemical study

Aim: Spleen is an organ of immune and hematopoietic system and involves extensive cellular interactions. Cell surface and extracellular matrix carbohydrates are important in cellular interactions (cell-cell and cell-matrix). Spleen is affected by the exposure to toxic agents. Crataegus (Hawthorn) species, which contains mainly flavonoid compounds, have such effects as antioxidant, cardioprotective, antiinflammatory and anticarcinogenic. In this study, the effects of Crataegus aronia var. dentata Browicz extracts on spleen were investigated via light microscopy. Materials and methods: The plant extract at concentration of 1% was given orally to rats for 6 weeks. At the end of the experimental period, tissues were fixed, processed and embedded in paraffin. 5 μm sections were stained with Hematoxylin and Eosin, Periodic acid-Schiff stain for histomorphology and Perl’s Prussian blue stain for hemosiderin. For lectin histochemistry, frozen sections were treated with three digoxigenin labelled lectins; peanut agglutinin (PNA) specific for galactose (1→3) N-acetylgalactosamine [Galβ(1→3)GalNAc]; Maackia amurensis leucoagglutinin (MAL) specific for sialic acidα(2→3)Galactose; Galanthus nivalis agglutinin (GNA) specific for terminal mannose residues. Results: In general, the histomorphology of the spleens and the accumulation of hemosiderin in macrophages were similar for both control and experimental groups. Lectins showed positive reactions in some cell cytoplasms and on the cell surfaces of both groups. A little more intensive GNA-positive reactions were observed on the cell surfaces. Conclusions: The plant extract at concentration of 1% did not lead to any alterations in some carbohydrate molecules and hemosiderin in the spleen

Histochemical investigation of the effects of Cyclamen graecum extracts on the rat lower gastrointestinal system

Aim: It is known that saponins, triterpenoid or steroidal glycosides, have effects on gastrointestinal system (GIS) and mucus. Cyclamen graecum , a tuberous plant belonging to the Primulaceae family, contains cyclamin as the main saponin. In the present study, it was aimed to investigate the effects of C. graecum tubers ethanolic extracts at the concentrations of 0.1 and 0.3 g/L on the rat lower GIS histomorphology and mucus intensity. Material-Method: In this study, rats were allocated to 3 groups, each comprising 5 rats [Group I: Control group, physiologic saline was administered; Group II: Experimental group, 0.1 g/L(aq) C. graecum extract was administered; Group III: Experimental group, 0.3 g/L(aq) C. graecum extract was administered]. C. graecum tubers ethanolic extracts were given orally to the rats during 10 weeks. At the end of the experimental period, Hematoxylin and Eosin (H&E), Periodic Acid-Schiff (PAS), Alcian Blue (AB) pH 1 and AB pH 2.5 histochemical staining techniques were employed on the tissue samples of small and large intestines in order to demonstrate the histomorphological structures, the neutral, sulphated and acidic mucosubstances, respectively. Results: In both experimental groups, small and large intestine histomorphology, and AB and PAS positive reaction intensities of neutral, acidic, and sulphated mucosubstances were similar to the control group. Discussion: It can be suggested that the C. graecum ethanolic extract has no effect on the rat lower GIS histomorphology and mucus intensity at the administered doses in our study

Immunohistochemical investigation of galectin-3 in the skin of mice applied with Origanum hypericifolium essential oil and irradiated with ultraviolet B

Aim: To demonstrate galectin-3-immunoreactivity in the undiluted essential oil of Origanum hypericifolium when applied to the ultraviolet B (UVB) irradiated skin of mice. Materials and methods: Female BALB/c mice were allocated to 4 groups, each comprising 6 mice (Group 1: control; Group 2: UVB irradiated control; Group 3: undiluted O. hypericifolium essential oil applied; Group 4: undiluted O. hypericifolium essential oil applied before UVB irradiation). One week prior to UVB irradiation, the undiluted O. hypericifolium essential oil was applied to the shaved dorsal skin of mice 3 times a week. Subsequently, the mice were irradiated 3 times per week with UVB (week 1: 50 mJ/cm2, week 2: 70 mJ/cm2, and weeks 3 and 4: 80 mJ/cm2) for 4 weeks. At the end of this period, immunohistochemical staining for galectin-3 was performed on frozen sections of skin specimens, and then they were photographed. Results: Numerous galectin-3-immunoreactive cells, which were considered to be immune system cells, were observed in the dermis of Group 3. Conclusion: It is suggested that undiluted O. hypericifolium essential oil may cause an increase in the galectin-3-immunoreactive cells. However, there is a need to research these findings with further molecular analyses. © TÜBİTAK

Glycohistochemistry of the lateral tympanic membrane in the syrinx of the denizli cock

The Denizli cock is a special race that has harmonious long crowing. In this study, the syrinx of the Denizli cock was studied by using glycohistochemical analyses. Paraffin sections were stained with Alcian blue (AB) at pH 1 and pH 2.5 for demonstration of sulfated and carboxylated carbohydrates, respectively. At AB-pH 1, the lateral tympanic membrane (LTM) and also the medial tympanic membrane (MTM) exhibited a weak reactivity, whereas they were stained intensely at AB-pH 2.5. The frozen sections were incubated with 5 digoxigenin-labeled lectins for lectin histochemistry: peanut agglutinin, Maackia amurensis leucoagglutinin, Galanthus nivalis agglutinin, Datura stramonium agglutinin, and Sambucus nigra agglutinin. In LTM, positive stainings were observed for the 5 lectins. These results suggest that LTM exhibits an intense reactivity with AB-pH 2.5, indicating carboxylated carbohydrates, and it also has terminal saccharide moieties consisting of β(1→3) linked galactose, α(2→3) linked sialic acid, mannose, β(1→4) linked galactose, and α(2→6) linked sialic acid. © Tübİtak

Cryobiology and Cryopreservation of Sperm

Low temperature has been utilized to keep living cells and tissues dormant but potentially alive for cryopreservation and biobanking with great impacts on scientific and biomedical applications. However, there is a critical contradiction between the purpose of the cryopreservation and experimental findings: the cryopreserved cells and tissues can be fatally damaged by the cryopreservation process itself. Contrary to popular belief, the challenge to the life of living cells and tissues during the cryopreservation is not their ability to endure storage at cryogenic temperatures (below −190°C); rather it is the lethality associated with mass and energy transport within an intermediate zone of low temperature (−15 to −130°C) that a cell must traverse twice, once during cooling and once during warming. This chapter will focus on (1) the mechanisms of cryoinjury and cryopretection of human sperm in cryopreservation, and (2) cryopreservation techniques and methods developed based on the understanding of the above mechanisms

Denizli yöresinde glukoz-6-fosfat dehidrogenaz enzim yetmezlikli bireylerde Akdeniz mutasyonu sıklığının ve eritrositlerdeki ince yapı değişikliklerinin araştırılması

Bu çalışmada, Denizli il merkezi ve ilçelerindeki G6PDAkdeniz 563T mutasyonu sıklığı ile, mutant eritrositlerde meydana gelen ince yapı değişiklikleri araştırılmıştır. Bu amaçla, orta dereceli okullarda yaşları 14-17 arasında değişen 918'i kız, 1032 si erkek olmak üzere toplam 1950 kişi taranmıştır. Kan örneklerinin kalitatif ve kantitatif ölçüm sonuçlarına göre 24 birey G6PD yetmezlikli bulunmuştur. PCR-RFLP çalışmasına, G6PD yetmezlikli olarak bulunan 24 birey ile, akrabalarından (anne-baba-kardeş) oluşan 12 birey olmak üzere, toplam 36 birey alınmıştır. Ekson VI ve VII'ye özgül primer çifti, sırasıyla AKA ve AKB kullanılarak 563T'nin amplifikasyon ve Mbo II restriksiyon endonükleaz enzimi ile kesim sonuçlarına göre, 24 bireyde G6PDAkdeniz 563T mutasyonu saptanmıştır. Çalışma grubundaki kalitatif ve kantitatif ölçüm sonuçlarına göre belirlenen 24 öğrencinin 19'unda (%79) G6PD Akdeniz 563T mutasyonu saptanmıştır. 19 bireyin 13'ü hemizigot, 2'si homozigot, 4'ü ise heterozigot olarak belirlenmiştir. Normal enzim aktivitesine sahip ancak heterozigot olan bireylerin saptanması kalitatif ve kantitatif yöntemlerin heterozigotlar için güvenilir olmadığını ortaya koymuştur. Kalitatif ve kantitatif ölçüm değerleri hemizigot bireylerinkine benzeyen ancak mutant bulunmayan bir erkek birey, bölgede başka mutasyonların varlığına işaret etmektedir. Normal ve mutant bireylerin eritrosit morfolojileri ışık mikroskobu (IM), taramalı (scanning) elektron mikroskobu (SEM), geçirimli (transmission) elektron mikroskobu (TEM) ile, yüzey anyonik yerlerin dağılışı da katyonik bir boya olan rutenyum kırmızısı (RR) kullanarak TEM mikroskobu ile karşılaştırılmıştır. IM ile toluidin mavisi ile boyanan mutant bireylere ait kesitlerde hipokromik ve hedef hücreler ayırt edilmiştir. Ayrıca hücrelerin içinde mavi mor renkte granüller şeklinde Heinz cisimcikleri görülmektedir. Mutant eritrositler SEM ile normal bikonkav disk şekillerinden oldukça farklı olarak, düzensiz şekillerde görülmektedir. Eritrositlerin çentikler şeklinde görülen kenarları TEM mikrograflarında hücre zarının sitoplazmaya doğru yaptığı geniş çöküntüler şeklinde ayırt edilmektedir. TEM mikrograflarında mutant eritrosit zarında erime ve zardan ayrılan parçaların görülmesi, hücrelerdeki yıpranmayı göstermesi bakımından anlamlıdır. Ayrıca, hücrelerin sitoplazmasında dejeneratif halkasal yapılar, elektronca yoğun kitleler ve vakuoller görülmektedir. Hemi/homozigot ve heterozigot bireylerin eritrositlerinde meydana gelen morfolojik değişiklikler bakımından fark bulunmamıştır.Tüm bireylere ait eritrositlerin yüzey anyonik yerleri RR ile pozitif reaksiyon vermiştir. Normal eritrosit yüzeyinde RR pozitif reaksiyon homojen olarak dağılım göstermiştir. Bununla birlikte reaksiyon mutant eritrosit yüzeyinde bölgesel olarak farklı yoğunluklarda ve heterojendir. PCR RFLP ve mikroskobik sonuçlar; Denizli yöresinde G6PD yetmezliğinin ileride de önemli bir sağlık sorunu olacağını, eritrosit genotipinde meydana gelen değişikliklerle eritrosit morfolojisi ve yüzey değişikliklerinin ilişkili olduğunu göstermektedir. Eritrositlerin makrofajlarca tanınarak zamanından önce dolaşımdan alınmasına neden olan glukokonjugatlardaki değişikliklerin araştırılması ve yörede başka mutasyonların saptanmasıyla ilgili olarak, yeni çalışmaların yapılması gerektiği ortaya çıkmıştır

Consanguineous marriages, and the relationship between consanguineous marriages and abortion, infant mortality in Acipayam, a rural area of Denizli, Turkey

Consanguineous marriages are common in Turkey. The rates and types of consanguineous marriages, and their relationship to abortions and infant mortality were investigated in Acipayam, a rural area of Denizli, Turkey. 795 families were interviewed using a pre-designed questionnaire. The total prevalence of consanguineous marriages is 17.5%. It is found that the first-cousin marriages are slightly higher (10.6%) than the second-cousin marriages (6.9%). Infant mortality and abortion rates in families were higher in the consanguineous marriages than non-consanguineous marriages (infant mortality 19.4%-> 3.5%, abortion 11.5%-> 2.6%, respectively). Also, positive relationship was found between the presence of infant mortality/abortions and consanguineous marriages (OR: 1.67; 95% CI: 1.12-2.49). This study emphasizes the significant effect of consanguineous marriages on the early mortality of the offspring in Acipayam in Denizli, Turkey

Glycohistochemical study on the Denizli cock testis

Denizli cock is a special race which has been breeding in Denizli province of Turkey for a long time. It is famous with its long, beautiful and harmanious crowing and beautiful appearance. In this study, the testis of the Denizli cock was investigated using histological and lectin histochemical methods by light microscopy. The distribution of lectin bindings in the seminiferous epithelium of testis was studied using five digoxigenin labelled lectins including Peanut Agglutinin (PNA), Datura stramonium Agglutinin (DSA), Galanthus nivalis Agglutimn (GNA), Maackia amurensis Leucoagglutinm (MAL) and Sambucus nigra Agglutinin (SNA). Some differences in lectin staining density in interstitial space, basement membrane, spermatogenic cells and sertoli cells have been detected. Interstitial space, basement membrane, spermatogenic cells with DSA and sertoli cells with GNA showed the most intense stainings. In spermatogenic cells, GNA, MAL and SNA displayed moderately stainings. However, they were weakly stained with PNA. In general, PNA, GNA, MAL and SNA stainings were similar moderate stainings in interstitial space and basement membrane. In sertoli cells, weak to moderate staining density of PNA, MAL and SNA but strong staining density of DSA according to stainings of PNA, MAL and SNA were observed. The results demonstrated the distribution of some glycoconjugates in the testis of Denizli cock. These lectin-bmding properties in Denizli cock testis were provided in reference to glycohistochemical investigations in birds. © Medwell Journals, 2011

A histochemical study of ultraviolet B irradiation and Origanum hypericifolium oil applied to the skin of mice

Ultraviolet (UV) rays cause skin damage. Chronic exposure to UV irradiation causes decreased collagen synthesis, degenerative changes in collagen bundles, accumulation of elastotic material and increased epidermal thickness. Origanum hypericifolium, an endemic Turkish plant, belongs to Lamiaceae family. The main constituents of its oil are monoterpenes including cymene, carvacrol, thymol and γ-terpinene. The effects of undiluted O. hypericifolium oil on UVB irradiated skin of mice were investigated histochemically. Four groups of female BALB/c mice, whose dorsal hair was shaved, were allocated as follows: non-UVB irradiated (Group 1), UVB-irradiated (Group 2), O. hypericifolium oil treated (Group 3), and O. hypericifolium oil treated and UVB irradiated (Group 4). Sections of dorsal skin samples were stained with Mallory's phosphotungstic acid hematoxylin for collagen fibers and Taenzer-Unna orcein for elastic fibers. Sections also were stained with hematoxylin and eosin to measure epidermal thickness. We observed intense staining of collagen and homogeneous, scattered thin elastic fibers in Group 1; scattered and weakly stained collagen and curled, amorphous, accumulate elastic fibers in Group 2; and intense staining of collagen in Groups 3 and 4. Accumulation of elastic fibers in the dermis was unremarkable in Groups 3 and 4. In Groups 3 and 4, O. hypericifolium oil treatment thickened the epidermis. Epidermal thickness was greatest in Group 4. We suggest that O. hypericifolium oil may block UVB induced alterations of collagen and elastic fibers, and increase epidermal thickness. © 2013 The Biological Stain Commission