Cervical mucus sialic acid content determines the ability of frozen-thawed ram sperm to migrate through the cervix.

The aim of this study was to investigate the properties and to functionally characterize the cervical mucus that modulates sperm transport through the cervix by using ewe breeds with a divergent pregnancy rate (Belclare and Suffolk; high and low, respectively) following cervical insemination using frozen-thawed semen. Sperm number, as well as sialic acid and fucose content in both the channels and in the lumen of different regions of the cervix were quantified in inseminated Belclare and Suffolk ewes. Expression of glycosyltransferase and MUC genes, glycosidase activity and sialic acid speciation in follicular phase cervical tissue and mucus were assessed. More spermatozoa were found in the cervical channels in the region closest to the cervical os in Belclare than Suffolk ewes (P < 0.05) and Suffolk ewes had a higher sialic acid content in the cervical channels than Belclare ewes (P < 0.05) in all regions of cervix. Suffolk ewes had significantly higher expression of FUT1, ST6GAL1 and MUC5AC than Belclare ewes. There was no difference between the breeds in glycosidase activity (P > 0.05). Levels of Neu5Ac were higher in Belclare than Suffolk ewes (P < 0.05) and levels of Neu5Gc was higher in Suffolk than Belclare ewes (P < 0.05). Competitive sperm penetration assays demonstrated that frozen-thawed sperm progression increased when cervical mucus was incubated with sialyllactose prior to a sperm penetration test (P < 0.05). These results suggest that the difference between Belclare and Suffolk ewes in sperm transport with frozen-thawed semen is due to the higher concentration of sialic acid within channels, which binds to spermatozoa and reduces their ability to traverse the cervix.

Siglec expression on the surface of human, bull and ram sperm.

Sialic acid (Sia) is a major constituent of both the sperm glycocalyx and female reproductive mucosal surface and is involved in regulating sperm migration, uterotubal reservoir formation and oocyte binding. Siglecs (sialic acid-binding immunoglobulin - like lectins) commonly found on immune cells, bind to Sia in a linkage- and sugar-specific manner and often mediate cell-to-cell interactions and signalling. Proteomic and transcriptomic analysis of human and bovine sperm have listed Siglecs, but to date, their presence and/or localisation on sperm has not been studied. Therefore, the aim of this study was to characterise the presence of Siglecs on the surface of bovine, human and ovine sperm using both immunostaining and Western blotting. Siglec 1, 2, 5, 6, 10 and 14 were identified and displayed both species- and regional-specific expression on sperm. Almost universal expression across Siglecs and species was evident in the sperm neck and midpiece region while variable expression among Siglecs, similar among species, was detected in the head and tail regions of the sperm. The possible role for these proteins on sperm is discussed.

Fertility and genomics: comparison of gene expression in contrasting reproductive tissues of female cattle.

To compare gene expression among bovine tissues, large bovine RNA-seq datasets were used, comprising 280 samples from 10 different bovine tissues (uterine endometrium, granulosa cells, theca cells, cervix, embryos, leucocytes, liver, hypothalamus, pituitary, muscle) and generating 260 Gbases of data. Twin approaches were used: an information-theoretic analysis of the existing annotated transcriptome to identify the most tissue-specific genes and a de-novo transcriptome annotation to evaluate general features of the transcription landscape. Expression was detected for 97% of the Ensembl transcriptome with at least one read in one sample and between 28% and 66% at a level of 10 tags per million (TPM) or greater in individual tissues. Over 95% of genes exhibited some level of tissue-specific gene expression. This was mostly due to different levels of expression in different tissues rather than exclusive expression in a single tissue. Less than 1% of annotated genes exhibited a highly restricted tissue-specific expression profile and approximately 2% exhibited classic housekeeping profiles. In conclusion, it is the combined effects of the variable expression of large numbers of genes (73%-93% of the genome) and the specific expression of a small number of genes (<1% of the transcriptome) that contribute to determining the outcome of the function of individual tissues.

Synthesis, analysis and biological evaluation of novel indolquinonecryptolepine analogues as potential anti-tumour agents.

A small library of cryptolepine analogues were synthesised incorporating halogens and/or nitrogen containing side chains to optimise their interaction with the sugar-phosphate backbone of DNA to give improved binding, interfering with topoisomerase II hence enhancing cytotoxicity. Cell viability, DNA binding and Topoisomerase II inhibition is discussed for these compounds. Fluorescence microscopy was used to investigate the uptake of the synthesised cryptolepines into the nucleus. We report the synthesis and anti-cancer biological evaluation of nine novel cryptolepine analogues, which have greater cytotoxicity than the parent compound and are important lead compounds in the development of novel potent and selective indoloquinone anti-neoplastic agents.

N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages.

Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans.

Improving bovine semen diluents: insights from the male and female reproductive tracts, and the potential relevance of cervical mucins.

The commercial applicability of bovine artificial insemination (AI) depends on the effectiveness of diluents for maintaining sperm fertility. Challenges faced by the AI industry due to recent advances in assisted reproduction, and the limitations inherent in using fresh and frozen-thawed sperm for AI, could be overcome with the development of better semen diluents. Research into the different microenvironments of bovine sperm as they progress towards maturity, capacitation and fertilisation is revealing various mechanisms that could be exploited to improve the formulation of semen diluents. These are reviewed here. A rationale for a more detailed investigation of bovine cervical mucus for factors that may allow further progress towards this goal are also discussed.

The antimicrobial screening of a Barbadian medicinal plant with indications for use in the treatment of diabetic wound infections / Tamizaje antimicrobiano de una planta medicinal Barbadense con indicaciones para su uso en el tratamiento de infecciones de heridas en diabéticos

OBJECTIVE: Diabetes mellitus is a chronic noncommunicable disease with high prevalence in the North American and Caribbean region. Diabetic Foot Syndrome which is an associated complication can lead to the development of wounds and ulcers which can become infected. Justicia secunda, a plant known locally in Barbados as Bloodroot used in folklore for wound healing, was selected to test its ability to aid diabetic wound healing by antimicrobial activity. It was therefore tested against the bacteria Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, and Enterococcus feacalis (clincal strain) which are commonly found in diabetic wounds. METHODS: The plant was collected by local users. Methanol and acetone extracts of the plant were prepared with use of soxhlet extraction. The antimicrobial activity was assessed with the use of a modified KirbyBaurer method. Concentrations of 200 mg/ml, 100 mg/ml, 10 mg/ml, and 1 mg/ml of the extract were used, with a standard ciprofloxacin 5 µg positive control, and a 5% dimethyl sulfoxide (DMSO) solution negative control. RESULTS: The J secunda methanol and acetone extracts with an extraction yield of 15.3% and 0.75%, respectively yielded no activity within the concentration range against the three strains of bacteria tested. In comparison with the positive control, relative inhibition zone diameter (RIZD) values of 0% resulted for both the negative control and the extracts, with the positive control having a value of 100%. CONCLUSION:The in vitro screen of the extracts prepared from J secunda, yielded no antimicrobial activity against the three strains of bacteria tested and therefore does not support the folklore claims by this mechanism of action.

OBJETIVO: La diabetes mellitus es una enfermedad crónica no transmisible, de alta prevalencia en la región de Norte América y el Caribea. El síndrome de pie diabético es una complicación asociada, que puede llevar al desarrollo de heridas y úlceras, con la consiguiente posibilidad de infección. Justicia segunda es una planta conocida localmente en Barbados como "bloodroot" (sanguinaria canadensis) y usada en la medicina folklórica para la curación de heridas. Esta planta fue seleccionada para analizar su capacidad de ayudar a curar las heridas de diabéticos por su actividad antimicrobiana. Por lo tanto, se la sometió a prueba frente a bacterias Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, y Enterococcus feacalis (cepa clínica) que normalmente se encuentran en las heridas del diabético. MÉTODOS: La planta fue proporcionada por usuarios locales. Usando un extractor Soxhlet, se prepararon extractos de metanol y acetona a partir de la planta. La actividad antimicrobiana se evaluó mediante el método de KirbyBauer modificado. Se usaron concentraciones de 200 mg/ml, 100 mg/ml, 10 mg/ml, y 1 mg/ml del extracto, con un control positivo de 5 µg de ciprofloxacina estándar, y un control negativo de una solución de dimetil sulfóxido (DMSO) al 5%. RESULTADOS: Los extractos de metanol y acetona de J secunda con un rendimiento de extracción de 15.3% y 0.75% respectivamente, no arrojaron actividad alguna dentro del rango de la concentración contra las tres cepas de bacterias sujetas a prueba. Comparado con el control positivo, el diámetro de la zona de inhibición relativa (RIZD) arrojó valores de 0%, tanto para el control negativo como para los extractos, con un valor de 100% para el control positivo. CONCLUSIÓN: El tamizaje in vitro de los extractos preparados de J secunda, no arrojó actividad antimicrobiana alguna contra las tres cepas de bacterias analizadas, y por consiguiente no sustenta la afirmación de la medicina folklórica en relación con este mecanismo de acción.

A comparison of the chemical constituents of Barbadian medicinal plants within their respective plant families with established drug compounds and phytochemicals used to treat communicable and non-communicable diseases.

OBJECTIVE: Barbados has a strong base in the practice of folklore botanical medicines. Consistent with the rest of the Caribbean region, the practice is criticized due to lack of evidence on the efficacy and safety testing. The objectives of this review article are i) to categorize and identify plants by their possible indications and their scientific classification and ii) to determine if the chemical constituents of the plants will be able to provide some insight into their possible uses in folklore medicine based on existing scientific research on their chemical constituents and also by their classification. METHOD: A review of the folklore botanical medicines of Barbados was done. Plants were primarily grouped based on their use to treat particular communicable and non-communicable diseases. Plants were then secondarily grouped based on their families. The chemical profiles of the plants were then compared to established drug compounds currently approved for the conventional treatment of illnesses and also to established phytochemicals. RESULTS: The extensive literature review identified phytochemical compounds in particular plants used in Barbadian folklore medicine. Sixty-six per cent of reputed medicinal plants contain pharmacologically active phytochemicals; fifty-one per cent of these medicinal plants contain phytochemicals with activities consistent with their reported use. CONCLUSION: Folklore botanical medicine is well grounded on investigation of the scientific rationale. The research showed that fifty-one per cent of the identified medicinal plants have chemical compounds which have been identified to be responsible for its associated medicinal activity. To a lesser extent, approved drug compounds from drug regulatory bodies with similar chemical structure to the bioactive compounds in the plants proved to validate the use of some of these plants to treat illnesses.

A comparison of the chemical constituents of Barbadian medicinal plants within their respective plant families with established drug compounds and phytochemicals used to treat communicable and non-communicable diseases / Una comparación de los elementos constitutivos químicos de las plantas medicinales Barbadenses dentro de sus respectivas familias de plantas, con los compuestos medicinales convencionales y los fitoquímicos usados para tratar las enfermedades comunicables y las no comunicables

OBJECTIVE: Barbados has a strong base in the practice of folklore botanical medicines. Consistent with the rest of the Caribbean region, the practice is criticized due to lack of evidence on the efficacy and safety testing. The objectives of this review article are i) to categorize and identify plants by their possible indications and their scientific classification and ii) to determine if the chemical constituents of the plants will be able to provide some insight into their possible uses in folklore medicine based on existing scientific research on their chemical constituents and also by their classification. METHOD: A review of the folklore botanical medicines of Barbados was done. Plants were primarily grouped based on their use to treat particular communicable and non-communicable diseases. Plants were then secondarily grouped based on their families. The chemical profiles of the plants were then compared to established drug compounds currently approved for the conventional treatment of illnesses and also to established phytochemicals. RESULTS: The extensive literature review identified phytochemical compounds in particular plants used in Barbadian folklore medicine. Sixty-six per cent of reputed medicinal plants contain pharmacologically active phytochemicals; fifty-one per cent of these medicinal plants contain phytochemicals with activities consistent with their reported use. CONCLUSION: Folklore botanical medicine is well grounded on investigation of the scientific rationale. The research showed that fifty-one per cent of the identified medicinal plants have chemical compounds which have been identified to be responsible for its associated medicinal activity. To a lesser extent, approved drug compounds from drug regulatory bodies with similar chemical structure to the bioactive compounds in the plants proved to validate the use of some of these plants to treat illnesses.

OBJETIVO: Barbados tiene una sólida base en la práctica de las medicinas botánicas del folklore De conformidad con el resto de la región caribeña, la práctica se critica debido a la falta de evidencia sobre la eficacia y la seguridad. Los objetivos de este artículo de revisión son (i) categorizar e identificar las plantas por sus posibles indicaciones y su clasificación científica, y (ii) determinar si los elementos constitutivos químicos de las plantas podrían proporcionar algún entendimiento de sus posibles usos en la medicina de folklore, a partir de la investigación científica existente sobre sus elementos químicos constitutivos, y también de su clasificación. MÉTODO: Se realizó una revisión de las medicinas botánicas folklóricas de Barbados. Las plantas fueron agrupadas principalmente a partir de su uso para tratar enfermedades comunicativas y no comunicativas específicas. Se agruparon entonces secundariamente sobre la base de sus familias. Se procedió entonces a comparar los perfiles químicos de las plantas con los compuestos medicinales establecidos, aprobados corrientemente para el tratamiento convencional de enfermedades. Igual procedimiento se realizó con los fitoquímicos establecidos. RESULTADOS: La extensa revisión de la literatura identificó compuestos fitoquímicos en plantas específicas usadas en la medicina folklórica de Barbados. Sesenta y seis por ciento de las plantas medicinales que gozan de reputación contienen elementos fitoquímicos farmacológicamente activos; el cincuenta y un por ciento de estas plantas medicinales contiene fitoquímicos con principios activos correspondientes al uso reportado. CONCLUSIÓN: La medicina botánica folklórica está bien cimentada en la investigación de los principios científicos. La investigación mostró que el cincuenta y un por ciento de las plantas medicinales identificadas tienen compuestos químicos que han sido identificados como responsables de la actividad medicinal asociada. En menor grado, los compuestos medicinales aprobados - provenientes de las instituciones que regulan los medicamentos con estructuras químicas similares a los compuestos bioactivos de las plantas - han servido como pruebas que validan el uso de algunas de estas plantas en el tratamiento de ciertas enfermedades.

The antimicrobial screening of a Barbadian medicinal plant with indications for use in the treatment of diabetic wound infections.

OBJECTIVE: Diabetes mellitus is a chronic non-communicable disease with high prevalence in the North American and Caribbean region. Diabetic Foot Syndrome which is an associated complication can lead to the development of wounds and ulcers which can become infected. Justicia secunda, a plant known locally in Barbados as Bloodroot used in folklore for wound healing, was selected to test its ability to aid diabetic wound healing by antimicrobial activity. It was therefore tested against the bacteria Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, and Enterococcus feacalis (clincal strain) which are commonly found in diabetic wounds. METHODS: The plant was collected by local users. Methanol and acetone extracts of the plant were prepared with use of soxhlet extraction. The antimicrobial activity was assessed with the use of a modified Kirby-Baurer method. Concentrations of 200 mg/ml, 100 mg/ml, 10 mg/ml, and 1 mg/ml of the extract were used, with a standard ciprofloxacin 5 microg positive control, and a 5% dimethyl sulfoxide (DMSO) solution negative control. RESULTS: The J secunda methanol and acetone extracts with an extraction yield of 15.3% and 0.75%, respectively yielded no activity within the concentration range against the three strains of bacteria tested. In comparison with the positive control, relative inhibition zone diameter (RIZD) values of 0% resulted for both the negative control and the extracts, with the positive control having a value of 100%. CONCLUSION: The in vitro screen of the extracts prepared from J secunda, yielded no antimicrobial activity against the three strains of bacteria tested and therefore does not support the folklore claims by this mechanism of action.

Social networking for adolescents with severe haemophilia.

Access to modern treatments allows adolescents with haemophilia to manage their haemophilia at home, with improved treatment outcomes and quality of life, but has reduced peer support and the potential for experiential learning from older peers. Social networking, aided by modern communication technologies, may offer health benefits through peer support. We sought to assess whether or not disease-specific social networking could benefit adolescents with severe haemophilia. A total of 150 adolescents (aged 10-18) with severe haemophilia A or B from 11 UK treatment centres or those who had attended focus groups to explore the potential for a social network designed specifically for their use were surveyed. Teenage boys with severe haemophilia in the UK who responded to an online and paper questionnaire (n = 47; 31% response rate) rarely knew of or socialized with others with haemophilia outside their families. Two-thirds of respondents said they would like to meet others. For 70% of boys, parents were the major source of information about haemophilia, yet more than half said they often had trouble finding answers to their questions. These boys frequently used online social networks to chat with friends. Adolescents with severe haemophilia frequently have limited contact with others and many wish to have greater contact. They may benefit from peer support and experiential learning gained through online social networking. The SixVibe restricted access social network is to be launched in 2011. It includes features designed to promote and facilitate the development of peer-to peer disease management skills for adolescents with severe haemophilia.

Glycoproteins and glycosidases of the cervix during the periestrous period in cattle.

The cervix and its secretions undergo biochemical and physical changes under the differential influences of estrogen and progesterone. These include changes in the glycoprotein profile of the endocervix and its secretions. A comprehensive survey of such changes in cervical epithelium and cervical secretions was performed on bovine samples throughout the periestrous period. Cervical tissue samples and swabs were collected from synchronized beef heifers that were slaughtered 1) 12 h after controlled intravaginal progesterone-releasing device (CIDR) removal, 2) 24 h after CIDR removal, 3) at the onset of estrus, 4) 12 h after the onset of estrus, 5) 48 h after the onset of estrus, and 6) 7 d after the onset of estrus. Histological staining with hematoxylin and eosin, periodic acid Schiff, Alcian blue, and high-iron diamine was carried out to map overall patterns of stored glycoproteins and tissue structure. Biotinylated lectins were also used to detect the presence and distribution of a range of saccharide structures. The activities of ß-galactosidase, α-L-fucosidase, ß-N-acetyl-hexosaminidase, and sialidase were measured in cervical swabs using specific substrates. The epithelial layer of the cervix exhibited dynamic changes in cellular hypertrophy and amounts of stored glycoprotein. The greatest content of neutral and acidic mucins was observed 48 h after onset of estrus (P < 0.05). Sialylated mucins predominated at the bases of cervical folds, whereas sulfated mucins were more abundant (P < 0.05) at their apices. The stained area of core mucin glycans changed (P < 0.05) in association with follicular versus luteal phases, whereas terminal glycans changed (P < 0.05) mainly at the time of estrus and shortly thereafter. The greatest activity of ß-galactosidase and sialidase was observed 12 h after onset of estrus, whereas ß-hexosaminidase and α-fucosidase peaked at the luteal time point (P < 0.05). Taken together, we suggest that the well-known changes in the endocervix and its secretions that are associated with the physiological modulation of sperm transport and function of the cervical barrier are, in part, driven by glycosylation changes.

Ascending placentitis in the mare: A review.

Ascending placentitis is a condition that occurs late in pregnancy when bacteria enter the sterile uterus from the lower reproductive tract. It leads to abortion or the birth of premature and weakened foals. Early detection and treatment of this condition is vital for ensuring the production of a viable foal.Mares with ascending placentitis often present in late term pregnancy with signs of premature udder development and premature lactation. There may be a vulvar discharge. Early detection of placental problems is possible using trans-abdominal or trans-rectal ultrasonography. Hormones such as progesterone and relaxin may be measured as indicators of foetal stress and placental failure. Postpartum foetal membranes may be thickened and contain a fibronecrotic exudate. The region most affected is the cervical star. Definitive diagnosis of ascending placentitis is by histopathological examination of the chorioallantoic membrane.Ideal treatment strategies are aimed at curing the infection and prolonging the pregnancy to as close to term as possible and consist of anti-microbials, anti-inflammatories and hormonal support.Swabs are taken from affected mares to determine antibiotic sensitivity and to aid in treatment of foals born from these mares which are at risk of becoming septic. If detected early enough, the chances of producing a viable foal are greatly increased.

A prospective, randomized, parallel group, controlled study of the effect of intensity of speech and language therapy on early recovery from poststroke aphasia.

OBJECTIVE: To examine whether the amount of speech and language therapy influences the recovery from poststroke aphasia. SETTING: A hospital stroke unit and community. DESIGN: A prospective, randomized controlled trial. INTERVENTION: Aphasic stroke patients were randomly allocated to receive 5 hours (intensive therapy group, n=51) or 2 hours (standard therapy group) of speech and language therapy per week for 12 consecutive weeks starting as soon as practicable after the stroke. Another 19 patients were recruited for 2 hours per week of therapy and were treated by National Health Service (NHS) staff (NHS group). OUTCOME MEASURE AND ASSESSMENT: The Western Aphasia Battery. Assessments were made blind to randomization at baseline and 4, 8, 12 and 24 weeks after the start of therapy. Data were analysed by intention to treat. RESULTS: The mean (SD) Western Aphasia Battery score at week 12 for the intensive, standard and NHS groups was 70.3 (26.9), 66.2 (26.2) and 58.1 (33.7), respectively. There was no treatment effect of intensive therapy (P > 0.05), but there was a statistically significant difference between the standard study and the NHS groups (P = 0.002 at week 12 and 0.01 at week 24). CONCLUSIONS: Intensive speech and language therapy (as delivered in this study) did not improve the language impairment significantly more than the ;standard' therapy which averaged 1.6 hours/week. The improvement in aphasia was least in patients who were in the NHS group. These patients received 0.57 (0.49) hours of therapy per week.

The rate and extent of improvement with therapy from the different types of aphasia in the first year after stroke.

OBJECTIVE: To examine the rate and extent of improvement from the different types of aphasia in the first year after stroke. DESIGN: A prospective longitudinal study. SETTING: A specialist stroke unit. PARTICIPANTS: Seventy-five aphasic patients with first-ever stroke. INTERVENTION: The type of aphasia was classified according to the criteria of the Western Aphasia Battery. The Western Aphasia Battery aphasia quotient was used to measure the initial severity and the rate and extent of improvement from aphasia. Assessments were made at baseline and 4, 8, 12 and 24 weeks later. RESULTS: The median percentage increase in the Western Aphasia Battery aphasia quotient was statistically higher in patients with Broca's aphasia than in the other groups at all weeks. Patients with Wernicke's aphasia had a significantly greater median percentage increase in their aphasia quotient than those with conduction and anomic aphasia at weeks 12 and 24, but less than patients with global aphasia at week 24. CONCLUSIONS: Patients with Broca's aphasia appear to have the best prognosis for improvement of language function in the first year of stroke. The extent of improvement in patients with global aphasia is better than that of patients with Wernicke's aphasia.

Interaction of Eimeria tenella with intestinal mucin in vitro.

The mucus gel layer overlying the gastrointestinal epithelium plays an important role in host-pathogen interactions. The initial interaction between the coccidian parasite Eimeria tenella and host cells of the intestinal epithelium must occur across this mucus interface. In this study, we examined the relationship between E. tenella and avian mucin, in particular the effect of purified intestinal regional mucin on parasite adherence and invasion in vitro. Secreted mucin from the chicken duodenum and cecum was purified by density gradient centrifugation and gel chromatography. Parasite invasion studies were performed in the Madin-Darby bovine kidney cell model. Eimeria tenella adherence to chicken duodenal mucin was detected, whereas adherence to cecal or bovine mucin was not shown. Parasite invasion into epithelial cells was not influenced by bovine mucin, whereas chicken mucin purified from the duodenum and cecum significantly inhibited invasion. Inhibition of E. tenella invasion into cells by mucin from the duodenum was marginally greater than that of the cecum, but this was not significant. This study demonstrated E. tenella interaction with native chicken intestinal mucin, which in turn inhibited parasite invasion into epithelial cells in vitro.

Internalization of near-infrared fluorescent dyes within isolated macrophage populations.

The development and application of microsensor technology has enhanced the ability of scientists to further understand various biological activities, such as changes in the intracellular environment after injury or toxic exposure. NIR microsensor technology may be useful in detecting the cellular injuries or adverse changes during the early onset period, allowing for the administration of therapies to initiate recovery. The development and use of Infrared (IR) and near infrared (NIR) dyes as biological micro-sensors due to their advanced spectral characteristics may be helpful. Three of the more useful NIR dye characteristics include the ability to minimize background interference by extraneous biological matrices, the ability to exhibit optimal molar absorptivity and quantum yields, and the ability to maintain normal cellular activity. Thus, the current study was designed to investigate the ability of selected NIR micro-sensor dyes to undergo cellular internalization, demonstrate intracellular NIR fluorescent signaling, and maintain normal cellular activity. The results demonstrate that the selected NIR micro-sensor dyes undergo cellular internalization. The presence of the dyes within the cells did not affect cell viability. In addition, these dyes demonstrate changes in absorbance and fluorescence after the immune cells were challenged with a stimulant. Moreover, critical cellular functions, such as tumor necrosis factor release and superoxide production were not compromised by the internalization of the fluorescent dyes. These data suggest that selected NIR micro-sensor dyes can undergo intracellular internalization within isolated macrophages without adversely affecting various parameters of normal cellular activity.

Identification of 9-O-acetyl-N-acetylneuraminic acid in normal canine pre-ocular tear film secreted mucins and its depletion in Keratoconjunctivitis sicca.

O-Acetylated sialic acids have been reported in many sialoglycoproteins where they mediate a variety of immune and other biological events. We have previously demonstrated that the protective mucus barrier on the surface of the canine eye contains sialoglycoproteins. We have also investigated the occurrence of O-acetylated sialic acids in these ocular mucins. Mucus aspirated from the surface of normal dog eyes and those with keratoconjunctivitis sicca (KCS) was fractionated into three pools by density gradient centrifugation. Sialic acids comprised 0.6-0.9% of the dry weight of the mucins isolated. The sialic acid profile in these pools was examined using HPLC. O-Acetylated sialic acids, mainly Neu5,9Ac2, were detected in normal animals and made up 10-30% of the total sialic acids detected. A doubling of the sialic acid content was found in KCS mucins, but the level of 9-O-acetylated sialic acid was reduced below 4% of total. Histological analysis of conjunctival tissue from normal and KCS dogs showed the presence of sialic acids, detected with the alpha(2-6) sialic acid-specific lectin Sambucus nigra, in the goblet cells and corresponding to the staining pattern for MUC5AC, the major ocular-secreted mucin gene product. In KCS animals a disruption of the normal pattern of conjunctival goblet cells was seen with preservation of the pattern of lectin binding observed in normal animals. Thus the data demonstrate the presence of mono-O-Acetylated sialic acids in normal canine ocular mucins and a loss of this population of sialic acids in dry eye disease in spite of a significant increase in total sialic acids in KCS mucin.

Quantification of the effects of melittin on liposome structure.

An optical technique, dual-polarization interferometry, has been used to examine lipid structures at the solid/liquid interface. Changes in the lipid structures, in real time, were examined as a consequence of challenging them with a peptide (melittin) that is known to induce liposome rupture. This work suggests that it should be possible to obtain a better understanding of the detail of the melittin rupture process.

High scores on the Western Aphasia Battery correlate with good functional communication skills (as measured with the Communicative Effectiveness Index) in aphasic stroke patients.

OBJECTIVE: To examine the correlation between an impairment-level and a functional-level assessment scale of aphasia. DESIGN: Prospective, longitudinal study. SETTING: A stroke rehabilitation unit. SUBJECTS: Sixty-seven aphasic acute stroke patients who were undergoing a multi-disciplinary rehabilitation programme, including conventional speech and language therapy (SLT). INTERVENTION: Patients were assessed on study entry and 4, 8, 12 and 24 weeks after the start of SLT. The language impairment was assessed with the Western Aphasia Battery (WAB) and the communicative functional limitation associated with aphasia was measured with the Communicative Effectiveness Index (CETI).Results. There was a statistically significant correlation between the two scales for all assessment periods (Pearson's r=0.71; P<0.01). CONCLUSION: The study suggests that in the acute and subacute stages of stroke the scores of WAB and CETI can be surmised from one another.