Rapid Detection of Avian Eimeria Species Using Denaturing Gradient Gel Electrophoresis

A denaturing gradient gel electrophoresis (DGGE) assay was developed to rapidly discriminate species of avian Eimeria. Amplification by PCR of the small subunit ribosomal RNA gene (approximately 1,600 nucleotides) with Eimeria genus-specific primers followed by cloning and sequencing allowed us to carry out phylogenetic analyses and identify clone sequences to species level in most cases. Clones were subsequently used to amplify a smaller fragment (approximately 120 nucleotides) suitable for DGGE. The fragments were separated on denaturing gradient gel and bands with unique migration distances were mixed to obtain an identification ladder. The identification ladder and PCR products obtained from DNA extracted from fecal samples from several poultry farms were compared. Applying the DGGE method in this study allowed a rapid differentiation of Eimeria species present in fecal samples collected from poultry farms

Structural and Functional Determinants of Rodent and Human Surfactant Protein A: A Synthesis of Binding and Computational Data

Surfactant protein A (SP-A) provides surfactant stability, first line host defense, and lung homeostasis by binding surfactant phospholipids, pathogens, alveolar macrophages (AMs), and epithelial cells. Non-primates express one SP-A protein whereas humans express two: SP-A1 and SP-A2 with core intra- and inter-species differences in the collagen-like domain. Here, we used macrophages and solid phase binding assays to discern structural correlates of rat (r) and human (h) SP-A function. Binding assays using recombinant rSP-A expressed in insect cells showed that lack of proline hydroxylation, truncations of amino-terminal oligomerization domains, and site-directed serine (S) or alanine (A) mutagenesis of cysteine 6 (C6S), glutamate 195 (E195A), and glutamate 171 (E171A) in the carbohydrate recognition domain (CRD) all impaired SP-A binding. Replacement of arginine 197 with alanine found in hSP-A (R197A), however, restored the binding of hydroxyproline-deficient rSP-A to the SP-A receptor SP-R210 similar to native rat and human SP-A. In silico calculation of Ca++ coordination bond length and solvent accessibility surface area revealed that the “humanized” R197A substitution alters topology and solvent accessibility of the Ca++ coordination residues of the CRD domain. Binding assays in mouse AMs that were exposed to either endogenous SP-A or hSP-A1 (6A2) and hSP-A2 (1A0) isoforms in vivo revealed that mouse SP-A is a functional hybrid of hSP-A1 and hSP-A2 in regulating SP-A receptor occupancy and binding affinity. Binding assays using neonatal and adult human AMs indicates that the interaction of SP-A1 and SP-A2 with AMs is developmentally regulated. Furthermore, our data indicate that the auxiliary ion coordination loop encompassing the conserved E171 residue may comprise a conserved site of interaction with macrophages, and SP-R210 specifically, that merits further investigation to discern conserved and divergent SP-A functions between species. In summary, our findings support the notion that complex structural adaptation of SP-A regulate conserved and species specific AM functions in vertebrates

Can the FUT 2 Gene Variant Have an Effect on the Body Weight of Patients Undergoing Bariatric Surgery? - Preliminary, Exploratory Study

Background: The FUT2 gene (Se gene) encoding the enzyme α-1,2-L-fucosyltransferase 2 seems to have a significant effect on the number and type of bacteria colonizing the intestines. Methods: In a group of 19 patients after bariatric surgery, the polymorphism (rs601338) of FUT2 gene was analyzed in combination with body mass reduction, intestinal microbiome (16S RNA sequencing), and short chain fatty acids (SCFA) measurements in stools. Results: Among the secretors (Se/Se polymorphism of the FUT2 gene rs601338, carriers of GG variant), correlations between waist-hip ratio (WHR) and propionate content and an increase in Prevotella, Escherichia, Shigella, and Bacteroides were observed. On the other hand—in non-secretors (carriers of GA and AA variants)—higher abundance of Enterobacteriaceae, Ruminococcaceae, Enterobacteriaceae, Clostridiales was recorded. Conclusions: The increased concentrations of propionate observed among the GG variants of FUT 2 may be used as an additional source of energy for the patient and may have a higher risk of increasing the WHR than carriers of the other variants (GA and AA)

Effect of Origanum chemotypes on broiler intestinal bacteria

Essential oils have been proposed as alternatives to antibiotic use in food animal production. This study evaluated 3 chemotypes of the Origanum genus, containing varying amounts of secondary metabolites carvacrol, thymol, and sabinene, in the broiler chicken diet. Aerial parts of Origanum vulgare L. (OL), O. vulgare L. ssp. hirtum (OH), and O. majorana (OM) were collected from a greenhouse located in the high altitude Sabana de Bogotá (Savanna of Bogotá) and O. vulgare L. ssp. hirtum (OG) produced and ground in Greece. Oregano essential oils (OEO) from these plants were obtained by steam distillation and analyzed by gas chromatography coupled to a mass spectrometer. Six treatments were evaluated: 200 mg/kg of OEO from OH, OL, and OM, 50 mg/kg of OEO from OG, 500 mg/kg of chlortetracycline, and without additives. Broiler chicks were maintained at 2,600 m above sea level, placed in brooder cages under a completely randomized design. Template DNA was isolated from duodenal, jejunal, ileal, and cecal contents in each group and bacterial 16S rDNA patterns were analyzed by denaturing gradient gel electrophoresis. Dendrograms of denaturing gradient gel electrophoresis band patterns revealed 2 main clusters, OEO-treated chicks and nontreated control chicks, in each intestinal segment. Band patterns from different gut compartments revealed major bacterial population shifts in the foregut (duodenum, jejunum, and ileum) compared with the hindgut (cecum and colon) at all ages evaluated (P \u3c 0.05). The OEO groups showed less shift (62.7% similarity coefficient) between these 2 compartments versus the control groups (53.7% similarity coefficient). A reduction of 59% in mortality from ascites was seen in additive-supplemented groups compared with the control group. This study represents the first work to evaluate the effects of the 3 main chemotypes of Origanum genus in broilers

Maganda at Malakas (Beautiful and Strong): The Filipino Woman as Artist across 19th Century and Contemporary Philippine Art

https://digitalcommons.hollins.edu/art_history_symposium_2020/1001/thumbnail.jp

Heparin modulates the 99-loop of factor IXa: Effects on reactivity with isolated Kunitz-type inhibitor domains

Reactivity of factor IXa with basic pancreatic trypsin inhibitor is enhanced by low molecular weight heparin (enoxaparin). Previous studies by us have suggested that this effect involves allosteric modulation of factor IXa. We examined the reactivity of factor IXa with several isolated Kunitz-type inhibitor domains: basic pancreatic trypsin inhibitor, the Kunitz inhibitor domain of protease Nexin-2, and the first two inhibitor domains of tissue factor pathway inhibitor. We find that enhancement of factor IXa reactivity by enoxaparin is greatest for basic pancreatic trypsin inhibitor (\u3e10-fold), followed by the second tissue factor pathway inhibitor domain (1.7-fold) and the Kunitz inhibitor domain of protease Nexin-2 (1.4-fold). Modeling studies of factor IXa with basic pancreatic trypsin inhibitor suggest that binding of this inhibitor is sterically hindered by the 99-loop of factor IXa, specifically residue Lys98. Slow-binding kinetic studies support the formation of a weak initial enzyme-inhibitor complex between factor IXa and basic pancreatic trypsin inhibitor that is facilitated by enoxaparin binding. Mutation of Lys98 to Ala in factor IXa results in enhanced reactivity with all inhibitors examined, whereas almost completely abrogating the enhancing effects of enoxaparin. The results implicate Lys98 and the 99-loop of factor IXa in defining enzyme inhibitor specificity. More importantly, these results demonstrate the ability of factor IXa to be allosterically modulated by occupation of the heparin-binding exosite

Use and non-use of intra-uterine device in the Philippines: Perspectives for the commercial sector

The public sector has been dispensing intra-uterine device for free to Filipino women for more than three decades now. Following the country\u27s movement towards contraceptive self-reliance, the government\u27s role would soon be diminished while the role of the private sector, which is already the major source of pills and condoms, would be expanded to include the provision of intra-uterine device. A study was carried out to review existing published quantitative data on the use of intra-uterine device and to collect qualitative evidence on women\u27s perceptions and experiences regarding their use or non-use of the method. Data were intended to inform the private sector about the method\u27s potential number of users and factors that influence women to use or not to use the contraceptive. The quantitative evidence was derived from the published reports of the National Demographic and Health Survey and the Family Planning Survey, while the qualitative data were drawn from focus group discussions involving current and former users of the intra-uterine device, intenders and limiters. Quantitative results indicate that the number of intra-uterine device users has been limited. Whereas its users grew by 0.8 percentage-points (3.3% to 4.1%) or by 40,000 new users in 2001-2003, the trend was not sustained; the rates have been declining from 4.1% in 2003 to 3.9% in 2004 and 3.7% in 2008. Since the device has the lowest discontinuation rate and is long lasting, the pool of users has comprised the same individuals through the years. Fortunately, there would be 0.391 million potential users. Qualitative findings suggest that women\u27s use and non-use of the intra-uterine device was guided by their perceptions of the advantages and disadvantages of the method, respectively. As a consequence of their actual experiences in using the method, their perceptions were enhanced, which subsequently influenced their continued or discontinued usage of the method. Thus, continued use was contingent on having positive experiences and discontinued use on having negative experiences of using the contraceptive. The extent in which women had positive or negative experiences depended on the quality of services they had received from health providers. The private sector has to market and promote the intra-uterine device as the next alternative method among contraceptive users and as a top choice among non-users. Whereas strategies must highlight the advantages of the method, these must also incorporate approaches to address the method\u27s disadvantages. Strategies must thus seek to address both perceptions and experiences of women. A study was carried out to review existing published quantitative data on the use of intra-uterine device and to collect qualitative evidence on women\u27s perceptions and experiences regarding their use or non-use of the method. Data were intended to inform the private sector about the method\u27s potential number of users and factors that influence women to use or not to use the contraceptive. The quantitative evidence was derived from the published reports of the National Demographic and Health Survey and the Family Planning Survey, while the qualitative data were drawn from focus group discussions involving current and former users of the intra-uterine device, intenders and limiters. Quantitative results indicate that the number of intra-uterine device users has been limited. Whereas its users grew by 0.8 percentage-points (3.3% to 4.1%) or by 40,000 new users in 2001-2003, the trend was not sustained; the rates have been declining from 4.1% in 2003 to 3.9% in 2004 and 3.7% in 2008. Since the device has the lowest discontinuation rate and is long lasting, the pool of users has comprised the same individuals through the years. Fortunately, there would be 0.391 million potential users. Qualitative findings suggest that women\u27s use and non-use of the intra-uterine device was guided by their perceptions of the advantages and disadvantages of the method, respectively. As a consequence of their actual experiences in using the method, their perceptions were enhanced, which subsequently influenced their continued or discontinued usage of the method. Thus, continued use was contingent on having positive experiences and discontinued use on having negative experiences of using the contraceptive. The extent in which women had positive or negative experiences depended on the quality of services they had received from health providers. \u27 The private sector has to market and promote the intra-uterine device as the next alternative method among contraceptive users and as a top choice among non-users. Whereas strategies must highlight the advantages of the method, these must also incorporate approaches to address the method\u27s disadvantages. Strategies must thus seek to address both perceptions and experiences of women

Heparin modulates the 99-loop of factor IXa: Effects on reactivity with isolated Kunitz-type inhibitor domains

Reactivity of factor IXa with basic pancreatic trypsin inhibitor is enhanced by low molecular weight heparin (enoxaparin). Previous studies by us have suggested that this effect involves allosteric modulation of factor IXa. We examined the reactivity of factor IXa with several isolated Kunitz-type inhibitor domains: basic pancreatic trypsin inhibitor, the Kunitz inhibitor domain of protease Nexin-2, and the first two inhibitor domains of tissue factor pathway inhibitor. We find that enhancement of factor IXa reactivity by enoxaparin is greatest for basic pancreatic trypsin inhibitor (\u3e10-fold), followed by the second tissue factor pathway inhibitor domain (1.7-fold) and the Kunitz inhibitor domain of protease Nexin-2 (1.4-fold). Modeling studies of factor IXa with basic pancreatic trypsin inhibitor suggest that binding of this inhibitor is sterically hindered by the 99-loop of factor IXa, specifically residue Lys98. Slow-binding kinetic studies support the formation of a weak initial enzyme-inhibitor complex between factor IXa and basic pancreatic trypsin inhibitor that is facilitated by enoxaparin binding. Mutation of Lys98 to Ala in factor IXa results in enhanced reactivity with all inhibitors examined, whereas almost completely abrogating the enhancing effects of enoxaparin. The results implicate Lys98 and the 99-loop of factor IXa in defining enzyme inhibitor specificity. More importantly, these results demonstrate the ability of factor IXa to be allosterically modulated by occupation of the heparin-binding exosite

Filipino women\u27s use of intra-uterine device and some contextual variables: Knowledge for commercial promotion

In its effort to achieve contraceptive self-reliance as a result of the gradual withdrawal of foreign donations of contraceptive supplies, the Philippines has been instituting \u27e user-pay scheme. This scheme, which requires women to pay for their contraceptives has already been implemented, albeit in differing levels, for condoms, and injectables. The IUD is next to be marketed as a commercial product. To guide the IUDs transition, a synthesis of relevant and existing quantitative and qualitative information in the Philippines was undertaken. Findings suggest that IUD has commercial prospects because it has potential groups of users, and it has unique attributes on which its commercial value can be packaged. To enhance its prospects, IUD’s service provision and marketing and promotion must be improve