Mastitis and fertility disorders in cows.

Udder inflammations, independently of significant losses in milk yield and alteration of its quality, can negatively affect cows' reproduction efficiency. Mastitis causes changes in many active constituents, both in milk and blood. Pathogenic changes in existing constituents and new active chemical compounds, generated during disease, can affect other organs, particularly the reproductive system and its mechanisms. Fertility disorders in mastitic cows are mostly connected with the activity of cytokines (especially TNF-alpha), cortisol, prostaglandin F2-alpha, reactive oxygen species (ROS), and classic inflammatory mediators. The successful treatment of mastitis as well as protection against udder infections should be considered important methods for prophylaxis of fertility disorders in cows. The first Polish trials indicated that injection of supportive drugs (antioxidants or lysozyme dimer or flunixin meglumine) to intramammarily treated cows can increase fertility in cows with mastitis.

Molecular characterization of Polish Prototheca zopfii mastitis isolates and first isolation of Prototheca blaschkeae in Poland.

Bovine mastitits caused by the colorless, yeast-like alga Prototheca zopfii is a serious and complex condition that results in heavy economic losses in the dairy industry, both through a substantial reduction in milk production and culling of infected animals. Based on the 18S rDNA sequence analysis, genotype-specific PCR assays have recently been developed to differentiate within the species P. zopfii three distinct P. zopfii genotypes (1-3), of which P. zopfii genotype 3 has been considered a new species P. blaschkeae sp. nov. The purpose of this study was to employ the newly-devised molecular approach for the detection of the two P. zopfii genotypes and P. blaschkeae sp. nov. among bovine mastitis isolates from Poland. This study is the first to provide molecular characterization of Polish P. zopfii mastitis isolates. It also gives the first description of bovine mammary protothecosis due to P. blaschkeae in Poland, as evidenced by genotypical, microbiological, and electron microscopy findings.

Selected biochemical blood compounds in cows with abomasum displacement.

The aim of this study was to compare some blood biochemical indicators in cows with displacement of abomasum (DA) which recovered or died after treatment. Examinations were performed on 60 multiparous cows with left (L) or right (R) displacement and on 15 healthy herdmates. Diagnosis was made by experienced practitioners on the basis of clinical examination. Surgical treatment was undertaken during the first 24 hours after diagnosis. Almost all animals (55 = 91.5%) became sick in the post parturient period (21 days p.p. on average) with the exception of 5 (8.3%) that became sick later. Blood samples were taken from each cow immediately before surgical procedure. Serum nonesterified fatty acid (NEFA), glucose (Glu), cholesterol (Chol), aspartate aminotransferase (AST), total bilirubin (Bil) and blood urea nitrogen (BUN) were measured. Sick animals were characterized by low mean values of Chol (< or = 2 mmol/l) and normal level of BUN (12-15 mg/dl), higher levels of NEFA (> 600 micromol/l) and Bil (> 22 micromol/l), higher activity of AST (> 100 U/l). Seven cows (11.67%) died after surgical correction and all others recovered. No significant differences in NEFA, Chol, AST, Bil and BUN levels were observed as dependent on the efficacy of treatment (survival, deaths). It was found that cows which died after surgical treatment were characterized by significant higher levels of glucose (5.05 mmol/l) compared to surviving cows (2.93 mmol/l).

Etiological agents of dairy cows' mastitis in western part of Poland.

The purpose of the study was to determine the etiological mastitis agents in cows from herds located in the western part of Poland in years 2003-2005. Bacteriological examinations of 18,713 samples taken from California Mastitis Test (CMT) positive quarters were performed according to standard methods. Some Gram-negative bacilli and Gram-positive cocci were examined with the use of API tests (Analytical Profile Index). It was found that 32.7% of samples that were CMT positive were culture negative and 9.3% of samples were contaminated. Streptococcus species (15.7%), coagulase negative staphylococci (CNS) (14.6%), Staph. aureus (8.6%), Gram-negative bacilli (4.0%) and Corynebacterium species (3.8%) were most frequently isolated. Escherichia coli (52.3%) dominated among Gram-negative bacilli followed by Klebsiella pneumoniae (4.1%), Pseudomonas aeruginosa (3.6%), Enterobacter cloacae (3.6%), Serratia marcescens (3.1%), Pasteurella multocida (3.1%), Acinetobacter lwofii (3.1%), and 26 other bacteria species. Stahylococcus xylosus (31.5%), Staph. hyicus (12.2%), Staph. haemolyticus (12.2%), Staph. sciuri (11.6%), Staph. chromogenes (8.8%), Staph. epidermidis (8.3%) and Staph. simulans (6.1%) were the most frequent CNS. Streptococcus uberis (50%), Str. dysgalactiae (19.7%), Str. acidominimus (6.6%) and Enterococcus faecalis (5.3%) were mostly found among CAMP-negative streptococci. An increase in frequency of mastitis caused by non-agalactiae streptococci, Staph. aureus, A. pyogenes and yeast-like fungi and a decrease in that produced by Str. agalactiae in 2005 as compared to years 2003 or 2004 were observed.

Influence of various treatment methods on bacteriological findings in cows with puerperal endometritis.

The aim of the study was to identify the species of microorganisms isolated from the uterus of healthy cows (control group) and cows affected with puerperal metritis (PM) before and after an experimental therapy with an immunomodulator and antibiotics versus commonly applied methods (antibiotic + beta-blocker or antibiotic + PGF2alpha). Examinations were carried out on 110 cows with PM in three farms with similar system of rearing and nutrition. The control group consisted of 21 cows without postpartum disturbances. Smears from the uterus were taken before treatment and then at 21st day of observation. Escherichia coli and other species of Enterobacteriaceae family were isolated from 48.2% of PM cows and 47.6% of healthy cows. The degree of Arcanobacterium pyogenes infection was statistically lower in healthy than in sick cows (9.5% versus 30.0%). Streptococcus sp. was isolated from 13.6% of PM cows and from 16% of control ones. Staphylococci were isolated from 10% of PM and from 30% of control cows. Other bacteria species were isolated from about 10% of the examined cows. The best elimination of infections (66%) was noted in cows treated with the intrauterine antibiotic in combination with PGF2alpha i.m. injection. Examination showed that species of bacteria in the postparturient uterus were similar in healthy and sick cows. However, Arcanobacterium pyogenes was isolated 3 times more often from the sick animals. This pathogen was identified in 11 cows out of 19 (57.9%) culled subsequently because of infertility after the metritis puerperalis was clinically cured.

The effect of some drugs injection to pregnant heifers on blood antioxidant status.

The purpose of the trial was to establish the effect of the injection of the lysozyme dimer or vitamins connected with Se on the activity of chosen antioxidant enzymes and the total antioxidant status in pregnant heifers. Examinations were carried out during winter season in one farm on 21 heifers aged 22-24 months. Between the 21st and 14st day before expected parturition, seven heifers were once i.m. injected with antioxidants (Vitamin A-600 000 i.u.; Vitamin D3-200 000 i.u.; Vitamin E-1.5 mg/kg b.w., Selenium-0.022 mg/kg b.w.), and the next seven animals with lysozyme dimer (Lydium-KLP) at a dose of 0.02 mg/kg b.w. versus 7 non-treated control animals. Blood samples were taken before injection and then in hour 24 and 72 after injection, and between, the 7th and 14th day after calving. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSHpx), glutathione reductase (GSHred) and total antioxidant status (TAS) were measured by colorimetric method with the use of Randox kits. The mean value of SOD activity 21-14 days before expected calving was 704.8 +/- 294.6 U/ml of whole blood, GSHpx 59222 +/- 23699 U/l of whole blood, GSHred 110.8 +/- 22.5 U/l and TAS 0.33 +/- 0.15 mmol/l of serum. These indicators did not change in the control group with the exception of a statistically insignificant decrease in SOD activity after parturition. Statistically significant increase in blood SOD activity was noted only in the first day after injection of vitamins combined with selenium. These antioxidants also caused an insignificant increase in blood GSHpx activity in 72 hour following the injection, and in the second week after calving (statistically significant). The injection of antioxidants or lysozyme dimer did not change the activity of blood GSHred. However, an increase in the TAS was found in hour 24 (non significant) and 72 (statistically significant) following the single injection of lysozyme dimer.

Field trials on the prophylaxis of intramammary infections in pregnant heifers.

The study was carried out in 5 farms on 174 pregnant heifers. Clinical examination of the udder and bacteriological tests of quarter secretion were performed between the 8th and 3rd week before parturition, and then the animals were divided into a control group (64 heifers) and 3 experimental groups and immediately treated. A group of 32 experimental heifers was injected once with antioxidants (Vitamin A--600,000 i.u.; Vitamin D3--200,000 i.u.; Vitamin E--1.5 mg/kg b.w., Selenium--0.022 mg/kg b.w., i.m.). The next group (26 heads) was intramammary infused with antibiotic DC product (cloxacillin). Heifers from last experimental group (52) were injected with lysosyme dimer in a single dose of 0.02 mg/kg b.w. Clinical and bacteriological examinations were made during the first week after calving. The presence of bacteria was found in secretion of 22.6-38.9% udder quarters in 56.2-71.2% of pregnant heifers. The number of infected quarters (cows) did not change distinctly in the first week after calving except the lysozyme dimer group, where a decrease by 30% was noted. The percentage of quarters with elevated somatic cell count was higher in antibiotic DC group and closely similar in the other groups. None of examined methods showed an acceptable prophylactic effect. Clinical mastitis cases during first week after parturition were mostly caused by Escherichia coli, Staph. chromogenes, Staph. simulans, Staph. aureus, Staph. hyicus, Str. uberis, Str. acidominimus and Enterococcus faecalis.

The use of some immunomodulators and non-antibiotic drugs in a prophylaxis and treatment of mastitis.

Antibiotics are widely used in treatment and prophylaxis of mastitis, but their antibacterial and therapeutic efficacy continues to decrease. New methods of prophylaxis of mastitis with the use of vaccines and biological response modifiers are reviewed. Prophylactic activity of staphylococcal vaccines should be investigated in the future because of difficulties in successful treatment of infections caused by these organisms. Vaccination with J5 bacterin (0111:B4) reduced duration of IMI and local signs of clinical coliform-mastitis. Some immunomodulators proved to be effective in treatment of mastitis during lactating period. Promising results were obtained with the use of giseng saponin, herbal gel, herbal extracts, propolis, lysosubtilin, antibacterial proteins, and lysozyme dimer especially.

Concentrations of triiodothyronine (T3), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in milk from healthy and naturally infected quarters of cows.

The effect of naturally acquired bacterial infection of the bovine udder on the activity of 5'-thyroxine monodeiodinase (5'-MD), and on the concentrations of the pro-inflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha in milk, from healthy (control) and inflamed quarters, was determined. The diagnostic procedure included history and clinical examination of the udder, macroscopic evaluation of secretions, the Californian Mastitis Test, determination of somatic cell counts and bacteriological examination of milk. It has been found that the milk triiodothyronine (T3) content and the 5'-MD activity from inflamed quarters were decreased when compared with controls. The decrease in the milk T3 from subclinical mastitic quarters was manifested when somatic cell counts were > 10(6) ml(-1). TNF-alpha was on average 2-fold higher in infected milk, and the concentration of IL-6 was unchanged. These results suggest that the decreased T3 content in mammary secretions during naturally occurring mastitis is associated with the severity of inflammation, increased TNF-alpha concentration and impaired enzymatic activity of 5'-MD.

Epithelial-myoepithelial carcinoma of the bronchus.

The authors describe an extremely rare case of epithelial-myoepithelial carcinoma of bronchial mucous glands involving lower lobe of the right lung, which was detected on a routine radiological examination in a 34-year-old woman, and then surgically resected.

Activation of the murine dihydrofolate reductase promoter by E2F1. A requirement for CBP recruitment.

The E2F family of heterodimeric transcription factors plays an important role in the regulation of gene expression at the G1/S phase transition of the mammalian cell cycle. Previously, we have demonstrated that cell cycle regulation of murine dihydrofolate reductase (dhfr) expression requires E2F-mediated activation of the dhfr promoter in S phase. To investigate the mechanism by which E2F activates an authentic E2F-regulated promoter, we precisely replaced the E2F binding site in the dhfr promoter with a Gal4 binding site. Using Gal4-E2F1 derivatives, we found that E2F1 amino acids 409-437 contain a potent core transactivation domain. Functional analysis of the E2F1 core domain demonstrated that replacement of phenylalanine residues 413, 425, and 429 with alanine reduces both transcriptional activation of the dhfr promoter and protein-protein interactions with CBP, transcription factor (TF) IIH, and TATA-binding protein (TBP). However, additional amino acid substitutions for phenylalanine 429 demonstrated a strong correlation between activation of the dhfr promoter and binding of CBP, but not TFIIH or TBP. Finally, transactivator bypass experiments indicated that direct recruitment of CBP is sufficient for activation of the dhfr promoter. Therefore, we suggest that recruitment of CBP is one mechanism by which E2F activates the dhfr promoter.

Detection and identification of a methanol-water complex by factor analysis of infrared spectra.

Fourier transform infrared absorption spectra, between 1850 and 2700 cm(-)(1), were used to study the intermolecular interactions between methanol and water. Digitized spectra obtained from methanol-water mixtures, with methanol mole fraction varying from 0 to 1.00, were subjected to factor analysis. Principal factor analysis indicated that three chemical factors were responsible for the data. Window factor analysis, a model-free method, was used to extract the concentration profiles of three species, which were identified as water, methanol, and a methanol-water complex. The profiles were used to deduce the stoichiometry of the complex, which was found to consist of one molecule of methanol and two molecules of water. The dissociation constant of the complex was determined to be 306 ± 33 M(2). These results differ from those reported by other investigators.

Thymomas and thymic carcinomas. Evaluation of the Müller-Hermelink classification.

In the present study the Müller-Hermelink classification was used to analyze 39 thymomas and 4 thymic carcinomas collected over 20 years at the Chair and Department of Pathomorphology CM UJ. Our series consisted of 10 medullary thymomas (25.6%), 13 mixed (33.3%), 7 organoid (17.9%) and 9 cortical thymomas (23.0%). The mean age of the patients was 50.3 and it was the highest in patients with mixed thymomas-58.4 years and the lowest in patients with organoid thymomas-38.9 years, the difference being statistically significant. In the present series women were predominant (24:15). Myasthenia gravis was observed in 10 patients (25.6%), 5 women and 5 men, in 7 of them it was associated with cortical and organoid thymomas. Medullary, mixed and organoid thymomas were usually of lower stage (I and II), having a better prognosis than cortical thymomas and thymic carcinomas. One organoid thymoma recurring after 7 years evolved into cortical thymoma. Two thymic carcinomas were diagnosed as anaplastic, low differentiated, one as sarcomatous and one as well differentiated squamous carcinoma. The mean age of the patients was 39.7 years. The prognosis in all the patients with thymic carcinoma was poor. Based upon personal experience the authors regard the Müller-Hermelink classification as useful, easy to apply, reproducible and providing important prognostic information.

Mature cystic teratoma of the mediastinum containing a carcinoid. A case report.

We report a case of mature mediastinal teratoma containing mature pancreatic tissue and carcinoid in a 57-year-old woman who underwent a successful surgical operation.

Pulmonary mucinous cystadenoma of borderline malignancy. A report of two cases.

Two cases of pulmonary mucinous cystadenoma of borderline malignancy with favourable prognosis have been reported. The discussion section contains a review of current opinion about differentiating these neoplasms from mucinous carcinomas.

Complexation between Copper(II) and Glycine in Aqueous Acid Solutions by Window Factor Analysis of Visible Spectra.

Window factor analysis (WFA), a self-modeling chemometric technique, is used to extract the concentration profiles of the complexes formed between Cu(II) and glycine (GlyH) during pH titration in the acid region. The visible absorption spectra of aqueous acid solutions, each containing 0.0020 M Cu(II) and 0.500 M glycine, are recorded at various pHs, ranging from 1 to 7. The concentration profiles and the spectral absorptivities of four copper species, Cu(2+), CuGlyH(2+), CuGly(+), and CuGly(2), are obtained. The spectral absorptivities of the complexes are in good agreement with those reported in the literature. The concentration profiles are used to determine the stepwise equilibrium constants of the three complexes. The reason why rank annihilation evolving factor analysis (RAEFA), a precursor technique, was not successful in resolving the spectral data is explained.

Effects of acetylsalicylic acid, chlormethine and glucose on cow's mammary gland.

It was found that 100 ml of 5% glucose solution, 200 ml of acetylsalicyclic acid or 200 ml of chlormethine introduced into udder quarter caused a temporary irritation of mammary gland in healthy cows after a single intramammary inlocation. The changes in milk manifested by an increase of somatic cells number, serum albumins level and LDH activity, and decreased lactose content disappeared after 48-60 hours.