Temporal dysregulation of genes in lamb testis cell during sheeppox virus infection.

The present study was aimed to elucidate the host-virus interactions using RNA-Seq analysis at 1 h and 8 h of post-infection of sheeppox virus (SPPV) in lamb testis cell. The differentially expressed genes (DEGs) and the underlying mechanisms linked to the host immune responses were obtained. The protein-protein interaction (PPI) network analysis and ingenuity pathway analysis (IPA) illustrated the interaction between the DEGs and their involvement in cell signalling responses. Highly connected hubs viz. AURKA, CHEK1, CCNB2, CDC6 and MAPK14 were identified through PPI network analysis. IPA analysis showed that IL-6- and ERK5-mediated signalling pathways were highly enriched at both time points. The TP53 gene was identified to be the leading upstream regulator that directly responded to SPPV infection, resulting in downregulation at both time points. The study provides an overview of how the lamb testis genes and their underlying mechanisms link to growth and immune response during SPPV infection.

Isolation and characterization of two lytic bacteriophages against Staphylococcus aureus from India: newer therapeutic agents against Bovine mastitis.

Bovine mastitis causes severe economic losses to dairy farmers. Staphylococcus aureus, is one of the most important pathogen implicated in etiology of clinical and subclinical mastitis in bovines. In view of increasing antimicrobial resistance alternatives to antibiotic therapy are much needed. The present decade has witnessed a renewed interest in phage based therapeutics and diagnostics. The present study, describes isolation and characterization of two lytic phages SAJK-IND and MSP against Staphylococcus aureus having a potential to be used in therapy against mastitis. SAJK-IND and MSP phages belonged to Myoviridae and Podoviridae families, respectively. TEM imaging of the two phages revealed an iscosahedral head. MSP phage has a short non contractile tail. SAJK-IND and MSP have a burst size of 44 ± 3 and 25 ± 5 PFU/ infected cell, respectively. SAJK-IND and MSP phages revealed Ì´ 12 and Ì´16 proteins, respectively on SDS-PAGE analysis. The lytic activity of the phages was specific for Staphylococcus aureus. SAJK-IND revealed 100% lytic activity against several strains of Staphylococcus aureus isolated from mastitis milk samples whereas, MSP had only 40% lytic activity. SAJK-IND phage genome was sequenced, assembled and deposited in Genbank under accession no MG010123.

Changing pattern of childhood blindness.

All children under the age of 16 who fulfilled the criteria of blindness and low vision as defined by WHO were included in the study. These children were recruited from 1990 to 2004 from all the Ophthalmology Departments of Ireland, National Council of Blind and Visually impaired. Data was collected from history, detailed ocular examination and investigations including CT, MRI, ultrasound and chromosomal analysis. The prevalence of blindness in 2004 was 0.05% compared to 0.02% in 1989. The aetiologies were divided in (1) genetic, (2) prenatal, (3) perinatal, (4) childhood categories. The genetic group was 33% of the total, (15.63%) had albinism (11%) had retinal dystrophies. The perinatal group of 27% optic nerve hypoplasia, structural anomalies like microphthalmos, anophthalmos comprised of 15.85% and cataract (5.47%). The perinatal group was 26%, cortical blindness (17.45%), ROP (5.5%) and the childhood group comprised of 12.4% of the total. The overall prevalence of childhood blindness and low vision was shown to have increased compared to 1989. The most significant observation was the decrease in childhood blindness due to ROP, owing to the early diagnosis and treatment and an increase in brain blindness due to cortical disease and disability. This has been shown in other studies and is due to increased survival of preterm neonate.

Qualitative and quantitative analysis of goat ovaries, follicles and oocytes in view of in vitro production of embryos.

Goat ovaries were collected from the slaughterhouse and categorized as right, left, corpus luteum (CL)-present and -absent group and evaluated on the basis of weight (g), length (cm), width (cm), number of follicles, follicles aspirated and number and state of cumulus-oocyte-complexes (COCs). Comparatively higher weight [(0.66+/-0.02) vs (0.64+/-0.02) g], length [(1.17+/-0.02) vs (1.11+/-0.02) cm] and width [(0.77+/-0.02) vs (0.74+/-0.02) cm] were found in right ovaries than those of left. On the other hand significantly (P<0.05) higher weight [(0.71+/-0.03) vs (0.64+/-0.01) g] and width [(0.76+/-0.03) vs (0.75+/-0.01) cm] were found in CL-present group than those of CL-absent group of ovaries. The left ovaries contained comparatively higher number of normal COCs [(1.06+/-0.09) per ovary] than right ovaries [(1.03+/-0.10) per ovary] and the similar trend was found in total number of follicles [(4.51+/-0.25) vs (4.30+/-0.23) per ovary] and follicles aspirated [(2.55+/-0.14) vs (2.52+/-0.12) per ovary]. But the total COCs per ovary was almost similar in both ovaries [right and left: (1.85+/-0.12) and (1.85+/-0.11) per ovary, respectively]. Higher number of total COCs [(1.87+/-0.09) vs (1.76+/-0.16) per ovary], total number of follicles [(4.45+/-0.19) vs (4.16+/-0.37) per ovary], follicles aspirated [(2.55+/-0.10) vs (2.48+/-0.21) per ovary] and normal COCs [(1.12+/-0.07) vs (0.76+/-0.14) per ovary] were found in CL-absent group than those of CL-present group of ovaries.

Aromatic amino acid biosynthesis and production of related compounds from p-hydroxyphenylpyruvic acid by rumen bacteria, protozoa and their mixture.

Aromatic amino acid biosynthesis and production of related compounds from p-hydroxyphenylpyruvic acid (HPY) by mixed rumen bacteria (B), protozoa (P), and their mixture (BP) in an in vitro system were quantitatively investigated. Microbial suspensions prepared from mature, fistulated goats fed Lucerne ( Medicago sativa) cubes and a concentrate mixture were anaerobically incubated at 39 degrees C for 12 h. Tyrosine (Tyr), phenylalanine (Phe), tryptophan (Trp) and other related compounds in both supernatants and hydrolyzates of all incubations were analyzed by HPLC. Large amounts of Tyr (27.0, 47.0 and 50.8% of disappeared HPY in B, P and BP, respectively) were produced from 1 mM HPY during a 12-h incubation period. The formation of Tyr in P was 1.8 and 1.6 times higher than those in B and BP, respectively. Appreciable amounts of Phe (3-12% of the disappeared HPY) and Trp (2-10% of the disappeared HPY) were also produced from HPY in B, P, and BP. Phe synthesis in B and P was almost similar but Trp synthesis in B was 1.8 times higher than that in P. The biosynthesis of both Phe and Trp from HPY in BP was higher than those in B plus P. A large amount of p-hydroxyphenylacetic acid (about 45% of the disappeared HPY) was produced from HPY in B which was 1.9 times higher than that in P. p-Hydroxybenzoic acid produced from HPY in P was 1.6 times higher than that in B. Considerable amounts of phenylpropionic acid, phenyllactic acid, and phenylpyruvic acid (2-6% of the disappeared HPY) were produced only in B.

Thin layer chromatographical detection of tyrosine produced from L-[U-(14)C]phenylalanine by ruminal microbes.

Thin layer chromatographical detection of tyrosine (Tyr) synthesized from L-[U-(14)C]phenylalanine (Phe) (1 mM) by rumen bacteria (B) and protozoa (P) collected from fistulated Japanese Goat was carried out. About 16 and 12% of the added Phe was converted to Tyr by B and P, respectively. Large amount of radioactivity in ether fractions indicated an abundant production of aromatic acids from Phe. Small amount of radioactivity found in CO(2) fractions implied an occurrence of considerable decarboxylation reaction(s) by rumen bacteria and protozoa.

The effect of anthropometric and socioeconomic factors on the racial difference in lung function.

African-Americans have lower lung function than whites. However, the relative contributions of body habitus and socioeconomic factors are unknown. To address this question, we analyzed data from 1242 white (806 women, 436 men) and 1084 African-American (696 women, 388 men) asymptomatic, nonsmoking adult participants of the third National Health and Nutrition Examination Survey (NHANES III). African-Americans were poorer, had larger FEV(1)/FVC and body mass index (BMI), but lower sitting height, FEV(1) and FVC than whites. Cross-sectional regression analyses using spirometric, anthropometric, and socioeconomic data were performed separately by sex to investigate racial differences in lung function. Sitting height accounted for 35-39% of the race difference in both sexes. Poverty index accounted for about 7.5% and 2.5% of the racial difference in women and men, respectively, whereas the effect of education accounted for about 2% in women and 4.7% in men. With further adjustment for BMI, we could account for only about half of the racial difference in FEV(1) and FVC. We conclude that the racial difference in lung function is only partially explained by a shorter upper body segment in African-Americans. Although low socioeconomic indicators are related to lower lung function, they explain only a small proportion of this racial difference.

The effect of gender on the relationship between body fat distribution and lung function.

Although abdominal obesity, as measured by waist-to-hip ratio (WHR), has long been recognized as a risk factor for metabolic and cardiovascular diseases, little is known about the effect of WHR on pulmonary function, especially in women. In this study of 1094 men and 540 women (18-102 years) from the Baltimore Longitudinal Study of Aging (BLSA), we examined the effect of WHR on forced expiratory volume in 1 s (FEV(1)). Cross-sectional analyses, after accounting for body mass index (BMI) and other variables, showed a strong inverse association of WHR with FEV(1) in men (beta = -1.338, P=.0001) but not in women. Furthermore, larger values of WHR were associated with greater reductions of forced vital capacity (FVC) in men (beta = -1.383, P =.0005) compared to women (beta = -0.679, P =.02). Thus, body fat distribution has independent effects on lung function that are more prominent in men than women.

Tryptophan biosynthesis and production of other related compounds from indole and L-serine by mixed ruminal bacteria, protozoa, and their mixture in vitro.

Tryptophan (Trp) biosynthesis and production of other related compounds from 1 mM each of indole (IND), L-serine (Ser), and IND plus Ser by mixed ruminal bacteria (B), protozoa (P), and their mixture (BP) in an in vitro system were quantitatively investigated. Ruminal microorganisms were anaerobically incubated at 39 degrees C for 12 h. Trp and other related compounds produced in both the supernatants and microbial hydrolyzates of the incubation were analyzed by HPLC. B, P, and BP suspensions were not able to produce Trp when incubated with only IND or Ser. Appreciable amounts of Trp (9.8, 3.1, and 6.6% of substrate) were produced from IND plus Ser after 12 h by B, P, and BP suspensions, respectively. Trp produced from IND + Ser in B was found only in the hydrolyzate, whereas it was found in the medium as a free form in P after a 12-h incubation period. Rumen bacteria and protozoa were separately demonstrated for the first time to produce Trp from IND plus Ser, and the ability of P to produce Trp from IND plus Ser was about one-third that of B in 12 h. Trp produced from IND plus Ser by B, P, and BP suspensions was simultaneously degraded into its related compounds, and, among them, indoleacetic acid (IAA) was a major product found in B. Production of IAA was 4.3, 0.3, and 3.2% of IND in 12 h by B, P, and BP suspensions, respectively. A small amount of skatole (SKT) (1.1 and 2.5% in B and BP, respectively) and p-cresol (CRL) (2.4 and 3.4% in B and BP, respectively) were also produced from IND plus Ser during 12-h incubation. P suspension produced no SKT or CRL from IND plus Ser in 12-h incubation. These results suggested for the first time that both rumen bacteria and protozoa have an ability to produce Trp from IND plus Ser, and the ability was higher in B than in P. The ratios of Trp produced from IND plus Ser to that from indolepyruvic acid by B, P, and BP were 1:3.4, 1:14.2, and 1:6.6 during 12-h incubation period. From these results, the degree of importance of producing Trp from IND plus Ser in the rumen was indicated.

Production of tyrosine and other aromatic compounds from phenylalanine by rumen microorganisms.

Rumen contents from three fistulated Japanese native goats fed Lucerne hay cubes (Medicago sativa) and concentrate mixture were collected to prepare the suspensions of mixed rumen bacteria (B), mixed protozoa (P) and a combination of the two (BP). Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h with or without 1 mM of L-phenylalanine (Phe). Phe, tyrosine (Tyr) and other related compounds in both supernatant and microbial hydrolysates of the incubations were analyzed by HPLC. Tyr can be produced from Phe not only by rumen bacteria but also by rumen protozoa. The production of Tyr during 12 h incubation in B (183.6 mumol/g MN) was 4.3 times higher than that in P. One of the intermediate products between Phe and Tyr seems to be p-hydroxyphenylacetic acid. The rate of the net degradation of Phe incubation in B (76.0 mumol/g MN/h) was 2.4 times higher than in P. In the case of all rumen microorganisms, degraded Phe was mainly (> 53%) converted into phenylacetic acid. The production of benzoic acid was higher in P than in B suspensions. Small amount of phenylpyruvic acid was produced from Phe by both rumen bacteria and protozoa, but phenylpropionic acid and phenyllactic acid were produced only by rumen bacteria.

Determinants of maximal inspiratory pressure. The Baltimore Longitudinal Study of Aging.

A variety of methods for subject selection and test procedures have been used for the determination of normal values and reference equations for maximal inspiratory pressure (MIP). In the cross-sectional study described here, we made MIP measurements on 668 men and women in the Baltimore Longitudinal Study of Aging (BLSA), using a standardized electronic procedure. Results were combined with spirometric and anthropometric measurements. After subjecting them to rigorous health screening, we analyzed a well-defined, healthy subgroup of 139 men and 128 women with a wide age range (20 to 90 yr), using multiple linear regression, for the purpose of determining the effect of age, other correlates, normal values, and gender-specific reference equations for MIP. The gender effect was strong, with the average MIP values of the men being about 30% higher than those of the women (101 cm H2O and 72 cm H2O, respectively). The reference equation for men is: MIP +/- standard error of the estimate (SEE) = 126 - 1.028 x age + 0.343 x weight (kg) +/- (22.4); and for women: MIP +/- SEE = 171 - 0.694 x age + 0. 861 x weight (kg) - 0.743 x height (cm) +/- (18.5). These equations may be used for the assessment of inspiratory muscle strength.

Quantitative determination of aromatic amino acids and related compounds in rumen fluid by high-performance liquid chromatography.

A rapid method for the quantitative determination of tyrosine (Tyr), phenylalanine (Phe), p-hydroxybenzoic acid (HBA), p-hydroxyphenylacetic acid (HPA), benzoic acid (BZA), p-hydroxyphenylpyruvic acid (HPY), phenylacetic acid (PAA), phenyllactic acid (PLA), tryptophan (Trp), indoleacetic acid (IAA), phenylpyruvic acid (PPY), phenylpropionic acid (PPA) and cinnamic acid (CNA) in goat rumen fluid was established by high-performance liquid chromatography (HPLC). The mobile phase used for isocratic elution was 50 mM sodium phosphate buffer (pH 6.5)-methanol (97:3, v/v). The flow-rate was 1.0 ml/min; column temperature 40 degrees C and compounds were monitored at 215 nm with a UV absorbance detector after injection of 10 microl of filtered rumen fluid. Analysis was completed within 40 min. The minimum detectable limits of quantification (microM) of these compounds were Tyr, 2; Phe, 3; HBA, 1; HPA, 2; BZA, 2; HPY, 8; PAA, 3; PLA, 4; Trp, 2; IAA, 2; PPY, 15; PPA, 8 and CNA, 4. Detectable levels of Tyr, Phe, HPA, BZA, HPY, PAA, PLA, Trp and PPA were found in the deproteinized rumen fluid of goat fed a haycube and concentrate mixture. PAA was the predominant compound before and after feeding. The concentrations of HPA, BZA, PAA, PLA and PPA in the goat rumen fluid increased after feeding, while the concentration of Tyr decreased. Phe, HPY and Trp were minor components at all times. PPY, IAA and CNA were not detected and HBA was not completely resolved in the goat rumen fluid.