Partial purification of pepsin from turkey proventriculus

In the present study, turkey proventriculus (stomach) was mixed with 10% NaCl (1:2 w/v) and extracted by centrifugation to produce crude extract. The partial purification of the extract was carried out by using Sephadex G-75 resin in gel filtration column chromatography. Crude extract (CE) and fractions obtained by gel filtration were analysed for milk-clotting activity (MCA), protein content, proteolytic activity (PA), and SDS-PAGE electrophoresis. The first 7 fractions did not have any activities; the fraction of 9, 10 and 11 had the most milk-clotting and proteolytic activities. Electrophoretic patterns proved that further purification steps should be applied for better results.     

Composition and in vitro antimicrobial activity of Populus buds and poplar-type propolis

The antibacterial activity of propolis has been widely investigated. Since reports dealing with antimicrobial activity of the origin of propolis are not available, this study was carried out aiming to analyse the in vitro antimicrobial activity of the methanol extracts of poplar type propolis and Populus (Populus nigra, P. alba, P. tremuloides) buds as its sources against standard strains of a panel of microorganisms by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The concentrations of the “poplar” phenolics were relatively high (4.5%) and some compounds typical for P. nigra such as pinobanksin and 4,3 acetyloxycaffeate were found in the propolis sample by GC-MS. The poplar type propolis and Populus bud exudates were found to inhibit most clinically important microorganisms in a wide spectrum including pathogenic yeasts but not Gram-negative bacteria.     

Radial flow hepatocyte bioreactor using stacked microfabricated grooved substrates

Bioartificial liver (BAL) devices with fully functioning hepatocytes have the potential to provide temporary hepatic support for patients with liver failure. The goal of this study was to optimize the flow environment for the cultured hepatocytes in a stacked substrate, radial flow bioreactor. Photolithographic techniques were used to microfabricate concentric grooves onto the underlying glass substrates. The microgrooves served to protect the seeded hepatocytes from the high shear stresses caused by the volumetric flow rates necessary for adequate convective oxygen delivery. Finite element analysis was used to analyze the shear stresses and oxygen concentrations in the bioreactor. By employing high volumetric flow rates, sufficient oxygen supply to the hepatocytes was possible without an integrated oxygen permeable membrane. To implement this concept, 18 microgrooved glass substrates, seeded with rat hepatocytes cocultured with 3T3-J2 fibroblasts, were stacked in the bioreactor, creating a channel height of 100 microm between each substrate. In this bioreactor configuration, liver-specific functions (i.e., albumin and urea synthesis rates) of the hepatocytes remained stable over 5 days of perfusion, and were significantly increased compared to those in the radial flow bioreactor with stacked substrates without microgrooves. This study suggests that this radial flow bioreactor with stacked microgrooved substrates is scalable and may have potential as a BAL device in the treatment of liver failure.     

Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease

Local delivery of IL-12 and GM-CSF to advanced primary tumors results in T- and NK-cell-dependent cure of disseminated disease in a murine spontaneous lung metastasis model. Post-therapy functional dynamics of cytotoxic T- and NK-cells were analyzed in primary and metastatic tumors to determine the specific roles of each subset in tumor eradication. Time-dependent depletion of CD8+ T and NK-cells demonstrated that CD8+ T-cells were critical to eradication of metastatic tumors within 3 days of treatment, but not later. In contrast, NK-cells were found to be essential to tumor regression for at least 10 days after cytokine delivery. Analysis of tumor-infiltrating lymphocyte populations in post-therapy primary tumors demonstrated that treatment resulted in the activation of tumor-associated CD8+ T-cells within 24 h as determined by IFNγ and perforin production. T-cell activity peaked between days 1 and 3 and subsided rapidly thereafter. Activation was not accompanied with an increase in cell numbers suggesting that treatment mobilized pre-existing T-effector/memory cells without inducing proliferation. In contrast, therapy resulted in a ≥3-fold enhancement of both the quantity and the cytotoxic activity of NK-cells in primary and metastatic tumors on day 3 post-therapy. NK-cell activity was also transient and subsided to pre-therapy levels by day 5. Depletion of CD4+ and CD8+ T-cells prior to treatment completely abrogated NK-cell infiltration into primary and metastatic tumors demonstrating the strict dependence of NK-cell recruitment on pre-existing T-effector/memory cells. Treatment failed to induce significant NK-cell infiltration in IFNγ-knockout mice establishing the central role of IFNγ in NK-cell chemotaxis to tumors. These data show that transient activation of tumor-associated T-effector/memory and NK-cells, but not long-term CD8+ T-cell responses, are critical to suppression of metastatic disease in this model; and reveal a novel role for pre-existing adaptive T-cell immunity in the recruitment of innate effectors to tumors. This work was supported by NIH/NCI grant R01-CA100656-01A1 to N.K.E.     

Analyzing the functions of large glycoconjugates through the dissipative properties of their absorbed layers using the gel-forming mucin MUC5B as an example

Glyconjugates such as mucins, proteoglycans, and polysaccharides form the structural basis of protective cell-surface layers. In particular gel-forming mucins define a zone between the epithelial cell layer and the environment. Such molecules are of extreme molecular weight 5-100 x 10(6) and size (Rg 20-300 nm). On this account their biochemistry is inseparable from their physical biochemistry. Combining laser light scattering and quartz crystal mass balance with dissipation methods (QCM-D) we have investigated the properties of the MUC5B mucin and its cognate fragments when bound to a hydrophobic surface. MUC5B forms the basis of gels responsible for the protection of the oral cavity, lung, and cervical canal surfaces. Here we show, by analyzing dissipative interactions of hydrophobic, gold, and polystyrene surfaces, with the intact MUC5B molecule, its reduced subunits, and glycosylated tryptic fragments (obtained after reduction), the formation of 40- to 100-nm-thick highly structured, hydrated interfaces. These interfaces are dominated in their geometry and dissipative properties by the negatively charged carbohydrate-rich domains of the molecule, the naked protein domains being responsible for attachment. These carbohydrate-rich surfaces have well-defined absorptive properties and permit the entry and entrapment of albumin-coated micro-beads into the absorbed layer at and below a size of 60 nm. However beads larger than 100 nm are completely excluded from the surfaces. These absorptive phenomena correlate with large changes in film dissipation and thus may not only be important in biological functions, e.g. binding viruses, but could also be informative to the surfaces (often ciliated) onto which such mucus films are attached.     

Utilizing peanut husk (Arachis hypogaea L.) in the manufacture of medium-density fiberboards

The main objective of this study was to investigate the potential of peanut husk (Arachis hypogaea L.) as a fiber–peanut mixture to produce fiberboards for general purposes. For panel production, the addition of peanut husk at various percentages to the wood fiber was the only variable. Panels produced utilizing peanut husk were compared to panels produced using 100% wood fiber. The chemical properties of peanut husk; holocellulose and lignin content, alcohol–benzene, hot and cold water, and dilute alkali (1% NaOH) solubility, were also determined. Results indicated that panels could be produced utilizing up to 30% peanut husk without affecting the usability of the panels. It was not possible to meet the minimum IB strength standards when peanut husk was added to the mixture. Higher additions resulted in panels having lower elastic and rupture moduli than the minimum requirements according to TS-EN standards.     

The genotoxic risk of underground coal miners from Turkey

A cytogenetic monitoring study was carried out on a group of workers from a bituminous coal mine in Zonguldak province of Turkey, to investigate the genotoxic risk of occupational exposure to coal mine dust. Cytogenetic analysis, namely sister chromatid exchanges (SCEs), chromosomal aberrations (CAs) and micronucleus (MN) tests were performed on a strictly selected group of 39 workers and compared to 34 controls matched for gender, age, and habit. Smoking and age were considered as modulating factors. Both SCE and CA frequencies in coal miners appeared significantly higher than in controls. Similarly, there was a significant increase in the frequency of total micronuclei in exposed group as compared to control group. The effect of smoking on the level of SCE and MN was significant in the control group. A positive correlation between the age and the level of SCE was also found in controls. The frequencies of both SCE and CA were significantly enhanced with the years of exposure. The results of this study demonstrated that occupational exposure to coal mine dust leads to a significant induction of cytogenetic damage in peripheral lymphocytes of workers engaged in underground coal mining.     

Changes in hemorheological parameters due to lead exposure in female rats

Lead is an ubiquitous metal in the environment that induces a broad range of physiological, biochemical and behavioral dysfunctions. The purpose of this study was to investigate its effects on blood parameters and blood viscosity. Female rats (14 Wistar-Albino type) were divided into a control and a lead exposed group. Both groups were fed with the same standard food, but lead acetate was added to the drinking water of the experimental group for 5 weeks. At the end of the experimental period, blood samples were drawn from the abdominal aorta of the anaesthetized animals. Hematocrit (Hct %), hemoglobin (Hb), and the number of erythrocytes were determined, blood viscosity was measured with a rotational viscometer, and the lead concentration in blood was analyzed by means of flame atomic absorption spectrometry. The erythrocyte count, Hb and Hct % of the lead exposed group were found to be significantly lower than in the control group (p<0.01). The blood viscosity level was significantly higher compared to the control group (p<0.01). It can be concluded that increased lead concentrations in blood impair certain hemorheological mechanisms and increase blood viscosity.     

Agronomical and chemical characterization of spearmint (Mentha spicata L.) originating in Turkey

The essential oil properties of spearmint (Mentha spicataL.), one of the most important spice plants, were studied and the essential oil components determined using gas chromatography. The essential oil content of wild-grown spearmint in the region was found to range from 1.00% to 2.00%, and two chemotypes were identified, one high in carvone (49.53-80.65%) and the other in pulegone (44.9-49.23%). Agronomic and essential oil properties of cultivated landraces ofM. spicata were also investigated under field conditions during the 1999 vegetation period. The examined spearmint landraces showed a great variability for each character studied, including yield and essential oil components. The crop was harvested twice during the vegetation period, and the essential oil content of the landraces varied from 0.90 to 2.70% in the first harvest and from 1.00 to 3.00% in the second one. Carvone was constantly present as the predominant essential oil in landraces, except for one sample, which was high in linalool (82.80%). Superior landraces with carvone contents were discovered; their maximum content reached 79.70% in the first cutting and 82.97% at the second cutting. The superior landraces were assayed for future improvement studies.     

First Homo erectus from Turkey and implications for migrations into temperate Eurasia

Remains of fossil hominins from temperate regions of the Old World are rare across both time and space, but such specimens are necessary for understanding basic issues in human evolution including linkages between their adaptations and early migration patterns. We report here the remarkable circumstances surrounding the discovery of the first fossil hominin calvaria from Turkey. The specimen was found in the Denizli province of western Turkey and recovered from within a solid block of travertine stone as it was being sawed into tile-sized slabs for the commercial natural stone building market. The new specimen fills an important geographical and temporal gap and displays several anatomical features that are shared with other Middle Pleistocene hominins from both Africa and Asia attributed to Homo erectus. It also preserves an unusual pathology on the endocranial surface of the frontal bone that is consistent with a diagnosis of Leptomeningitis tuberculosa (TB), and this evidence represents the most ancient example of this disease known for a fossil human. TB is exacerbated in dark-skinned peoples living in northern latitudes by a vitamin D deficiency because of reduced levels of ultraviolet radiation (UVR). Evidence for TB in the new specimen supports the thesis that reduced UVR was one of the many climatic variables presenting an adaptive challenge to ancient hominins during their migration into the temperate regions of Europe and Asia.