Ductal Lavage for Patients With Nonlactational Mastitis: A Single-Arm, Proof-of-Concept Trial.

BACKGROUND: Surgery, steroids, and/or observations alone have been proposed for patients with nonlactational mastitis (NLM), but most of these studies were retrospective. The optimal treatment for these patients remains unclear. This prospective, single-arm, proof-of-concept trial aimed to evaluate the feasibility and safety of ductal lavage as a novel treatment for patients with NLM. METHODS: Eligible patients with NLM received an intraductal infusion of corticosteroids and antimicrobial agents and returned the next day for a breast massage. This cycle was repeated for 2 wk, and we followed up these patients for 1 y. Patients did not receive surgery or steroids after ductal lavage. The primary endpoint was the time to complete response (CR). RESULTS: This trial included 32 patients with a median (range) age of 32 (20-53). Skin erythema and tenderness were the major symptoms. The median (range) visual analog score was 5 (0-9). There were 21 (65.6%), 4 (12.5%), and 7 (21.9%) patients diagnosed as idiopathic granulomatous mastitis, periductal mastitis, and unspecific NLM, respectively. During the ductal lavage, the median (range) number of cannulated ducts at first attempt was 5 (3-8). Ductal lavage significantly reduced the visual analog score and mastitis score (M-score) (P < 0.01). Within a median follow-up of 15.6 mo, 93.8% (30/32) of patients achieved CR. The median (range) time to CR was 6 (0.5-21) mo. Three patients (10.0%) relapsed. No adverse events associated with ductal lavage were observed. CONCLUSIONS: Ductal lavage for patients with NLM is feasible and safe, and a definitive randomized controlled trial for further investigation is warranted. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02794688.

The effect of subclinical mastitis on the concentration of immunoglobulins A, G, and M, total antioxidant capacity, zinc, iron, total proteins, and calcium in she-camel blood in relation with pathogens present in the udder.

The present study aims to evaluate the amount of immunoglobulins A, G, and M in she-camel blood serum in relation with the presence of pathogens in the udder, and to compare the antioxidative capacity and the concentration of zinc, iron, total proteins, and calcium. Milk and blood samples from she-camels from south Jordan were taken; according to milk bacteriological examination, the animals were divided into two groups: (Gm) which contained samples of milk contaminated with bacteria and (Gh) which contained uncontaminated milk samples. Milk and blood were sampled from 30 females and examined for the concentration of immunoglobulins A, G, and M and for the presence of pathogens in milk. Total antioxidant capacity, zinc, iron, total proteins, and calcium concentrations in blood were determined. Milk samples were checked for the presence of pathogens. She-camels for the study were of similar age and productivity in the middle stage of lactation. It was determined that the presence of pathogenic bacteria infecting the udder quarters had considerably influenced the values of immunoglobulins G, A, and M, total antioxidant capacity, and zinc and total protein concentration (p < 0.05) in blood serum; no significant difference in iron and calcium concentration was determined. Subclinical mastitis has a crucial role in increasing the concentration of immunoglobulins in serum; some parameters measured in blood (zinc, total antioxidant capacity, total proteins) could be indicative for the presence of inflammation in she-camels.

ANGPTL3 is part of the machinery causing dyslipidemia majorily via LPL inhibition in mastitis mice.

Previous investigations have shown that inflammation induces changes in lipid and lipoprotein metabolism, and increased expression of angiopoietin-like protein 3 (ANGPTL3) contributes to the development of dyslipidemia. Here we investigated whether there is a correlation between increased ANGPTL3 expression and dyslipidemia in mastitis mice. Thirty mice were divided into two groups: control group and Staphylococcus aureus (S. aureus)-induced mastitis mice group. Changes in the levels of blood lipids [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C)]; activity of myeloperoxidase (MPO); concentrations of plasma inflammation biomarkers [interferon-γ (IFNγ), tumor necrosis factor α (TNFα), and interleukin-1α (IL-1α)]; concentration of plasma ANGPTL3 protein; lipoprotein lipase (LPL) activities in postheparin plasma; expressions of hepatic N-acetylgalactosaminyltransferase 2 (GALNT2), hepatic ANGPTL3 and adipose LPL were determined. The major results indicated specific pathological mammary tissue changes, elevated MPO activity, reduced GALNT2 mRNA expression, elevated ANGPTL3 mRNA and protein expression and reduced LPL mRNA and protein expression. In plasma samples the S.aureus infused mice displayed elevated ANGPTL3 protein concentration, TG, TC and LDL-C levels, and reduced postheparin LPL activities and HDL-C level. The data suggests that ANGPTL3 is part of the machinery causing dyslipidemia majorily via LPL inhibition in mastitis mice.

Effects of Jin-Ying-Tang on Staphylococcus aureus-induced mastitis in rabbit.

The present study was performed to investigate the effects of Jin-Ying-Tang (JYT), a Chinese herbal formula containing Lonicera japonica, Herba taraxaci, Fructus trichosanthis, Fructus forsythia, Radix et rhizoma rhei, Astragalus membranaceus, Angelica sinensis, on rabbit mastitis induced by Staphylococcus aureus. Suckling rabbits were challenged with 1.5 × 10(7) colony forming unit (CFU) of S. aureus at the base of the third pair teats, and they were treated and pretreated with JYT to detect the formula effects. The results showed that JYT could reduce the occurrence of Staphylococcal mastitis in rabbit model. To further investigate the action mechanism of JYT, we examined the leukocyte counts and inflammatory mediator levels such as TNF-α and IL-6 in blood and infected tissue. From histological study and blood analysis, we found that JYT could suppress leukocyte infiltration in infected mammary gland tissue and significantly inhibit the total leukocyte counts and lymphocytes (LYM), monocytes (MON) and granulocytes (GRA) fractions of leukocyte counts in blood. Enzyme-linked immunosorbent assay (ELISA) results showed JYT significantly decreased the TNF-α and IL-6 concentrations in serum and mammary gland. The analysis of these data suggested that JYT effectively inhibited inflammatory responses to reduce the occurrence of mastitis in rabbit model.

Combining contrast-enhanced ultrasound and blood cell analysis to improve diagnostic accuracy of plasma cell mastitis.

Plasma cell mastitis is a benign suppurative disease of the breast, lack of specific clinical manifestations, which is easy to be misdiagnosed and mistreated, often confused with mastitis, breast cancer (BC), and other diseases. Thus, we aimed to establish a combined model of promoting diagnostic accuracy of plasma cell mastitis by contrast-enhanced ultrasound (CEUS) patterns and routine blood cell analysis. Eighty-eight plasma cell mastitis, 91 breast cancer, and 152 other benign breast diseases' patients grouped according to pathological diagnosis underwent CEUS and blood cell analysis examination; 100 healthy female donors were involved. All the plasma cell mastitis and breast cancer patients presented hyperenhancement of CEUS breast lesions compared with others. The majority of plasma cell mastitis (65/88) showed perfusion defect of CEUS patterns with smooth edge (56/65) and multiple lesions (49/65); in contrast, fewer breast cancer patients (30/91) displayed perfusion defect. White blood cell count (WBC), neutrophils, and neutrophils/lymphocytes ratio of blood cell analysis in plasma cell mastitis patients increased significantly compared with other patients (P < 0.0001). Combining perfusion defect of CEUS patterns and WBC yielded an area under the receiver operating characteristic curve of 0.831, higher than single 0.720 and 0.774, respectively. The cut-off value of WBC (7.28 × 109/L) helped remaining 65.2% (15/23) atypical cases to be correctly diagnosed as plasma cell mastitis, not misdiagnosed as breast cancer. In conclusion, CEUS presented a clear perfusion defect pattern of plasma cell mastitis lesion for the first time. A precise WBC by routine blood cell analysis test can assist CEUS examination in the differential diagnosis of plasma cell mastitis and breast cancer. It is a promised combination for laboratory diagnostic of PCM.

Development and validation of an ELISA for the quantification of bovine ITIH4 in serum and milk.

Inter alpha trypsin inhibitor heavy chain 4 (ITIH4) is a serum protein belonging to the Inter alpha trypsin inhibitor (ITI) family, which was previously characterized by our group as a new APP in cattle. This protein was firstly described in pigs where is known to be a major acute phase protein, also denominated Pig-MAP. Increases of ITIH4 of up to 12 times the pre-infection values were previously reported in the serum of heifers with experimentally induced summer mastitis. ITIH4 was detected in the milk of cows with mastitis by western blot, but the method previously used to quantify this protein, radial immunodiffusion, was not sensitive enough to quantify it in milk samples. In this study we developed an ELISA method which allows the quantification of bovine ITIH4 in serum and milk samples. Previously developed antibodies were used to perform the assay, including anti bovine ITIH4 polyclonal antibodies and a monoclonal antibody against pig ITIH4 that also recognizes the bovine homologous protein. The ELISA developed showed an adequate precision, with inter and intra- assay coefficients of variation lower than 10% for serum and milk samples. The assay keeps linearity under dilution for both serum and milk samples. A good agreement was observed between the values measured by ELISA and radial immunodiffusion in serum samples.

Differential quantitative proteomics study of experimental Mannheimia haemolytica mastitis in sheep.

Objective was the differential quantitative proteomics study of ovine mastitis induced by Mannheimia haemolytica; clinical, microbiological, cytological and histopathological methods were employed for confirmation and monitoring. Proteins were separated by two-dimensional gel electrophoresis (2-DE) for all samples and differentially abundant proteins were identified by mass spectrometry; comparisons were performed with pre- (blood, milk) and post- (milk of contralateral gland) inoculation findings. Animals developed mastitis, confirmed by isolation of challenge strain and increase of neutrophils in milk and by histopathological evidence. In blood plasma, 33 differentially abundant proteins (compared to findings before challenge) were identified: 6 with decrease, 13 with new appearance and 14 with varying abundance. In a post-challenge milk whey protein reference map, 65 proteins were identified; actin cytoplasmic-1, beta-lactoglobulin-1/B, cathelicidin-1 predominated. Further, 89 differentially abundant proteins (compared to findings before challenge) were identified: 18 with decrease, 53 with new appearance, 3 with increase and 15 with varying abundance; 15 proteins showed status changes in blood plasma and milk whey. Differential abundance from inoculated and contralateral glands revealed 74 proteins only from the inoculated gland. Most differentially abundant proteins in milk whey were involved in cell organisation and biogenesis (n = 17) or in inflammatory and defence response (n = 13). SIGNIFICANCE: The proteomes of blood and milk from ewes with experimental mastitis caused by Mannheimia haemolytica and the differential proteomics in sequential samples after challenge are presented for the first time. This is the first detailed proteomics study in M. haemolytica-associated mastitis in ewes. An experimental model fully simulating natural mastitis has been used. Use of experimentally induced mastitis minimised potential variations and allowed consistency of results. The study included evaluation of changes in blood plasma and milk whey. Protein patterns have been studied, indicating with great accuracy changes that had occurred as part of the disease process and development, during the acute phase of infection. Relevant protein-protein interactions were studied. The entirety of proteomics findings has suggested that affected ewes had mounted a defence response that had been regulated by many proteins (e.g., cathelicidins, haptoglobin, serum amyloid A) and through various pathways (e.g., acute phase response, binding and transporting significant ions and molecules); these were interdependent at various points. Potential biomarkers have been indicated for use in diagnostic assays of mastitis.

[Mastitis in infancy].

Data on epidemiology, etiology, pathogenesis, clinical picture, diagnostics and treatment of mastitis in infancy are presented. Higher frequency of mastitis in children was detected in 1986-2005 as compared with 1970-1985, less frequency of mastitis in newborn infants being observed and considerable growth of it in adolescent girls. Probable causes of the phenomenon are discussed.

Assessment of Poly(ADP-ribose) Polymerase1 (PARP1) expression and activity in cells purified from blood and milk of dairy cattle.

Poly(ADP-ribosyl)ation (PAR) is a post-translational protein modification catalysed by enzyme member of the poly(ADP-ribose) polymerases (PARPs) family. The activation of several PARPs is triggered by DNA strand breakage and the main PARP enzyme involved in this process is PARP1. Besides its involvement in DNA repair, PARP1 is involved in several cellular processes including transcription, epigenetics, chromatin re-modelling as well as in the maintenance of genomic stability. Moreover, several studies in human and animal models showed PARP1 activation in various inflammatory disorders. The aims of the study were (1) to characterize PARP1 expression in bovine peripheral blood mononuclear cells (PBMC) and (2) to evaluate PAR levels as a potential inflammatory marker in cells isolated from blood and milk samples following different types of infection, including mastitis. Our results show that (i) bovine PBMC express PARP1; (ii) lymphocytes exhibit higher expression of PARP1 than monocytes; (iii) PARP1 and PAR levels were higher in circulating PBMCs of infected cows; (iv) PAR levels were higher in cells isolated from milk with higher Somatic Cell Counts (SCC > 100,000 cells/mL) than in cells from milk with low SCCs. In conclusion, these findings suggest that PARP1 is activated during mastitis, which may prove to be a useful biomarker of mastitis.

Blood ionized calcium levels and acute-phase blood glucose kinetics in goats after intramammary infusion of lipopolysaccharide.

The aim of this study was to determine the blood ionized calcium (Ca) levels and acute-phase blood glucose kinetics in goats with mastitis induced by an intramammary challenge of lipopolysaccharide (LPS). Five goats were subjected to intramammary challenge of either LPS (10 µg) or saline (control). Some clinical manifestations (rectal temperature, pulse rate, respiration rate, ruminal motility, physical activity, and dehydration) were observed, and blood was collected for the measurement of several parameters [ionized and total Ca levels, blood glucose level, pH, and white blood count (WBC)] at 0 (just before challenge), 1-4, 6, 8, 12 and 24 hr post-challenge in both the LPS and control phases. Milk was collected at 0 (just before challenge), 4, 8, 12 and 24 hr post-challenge to measure the somatic cell count (SCC) and N-acetyl-beta-D-glucosaminidase (NAGase) activity. In the LPS phase, increased rectal temperature, significantly decreased ionized Ca and total Ca levels and WBCs were observed compared with those at 0 hr, although there were no differences in all parameters between phases. LPS infusion significantly increased SCCs in milk and NAGase activity. The present results demonstrated that, during the acute phase of mastitis induced by intramammary challenge by LPS at a concentration sufficient to cause general symptoms in goats, a decreased blood ionized Ca level occurs, but not hypoglycemia.

The Antisecretory Factor in Plasma and Breast Milk in Breastfeeding Mothers-A Prospective Cohort Study in Sweden.

Inflammation and infection postpartum threaten the mother and her infant. Human milk provides a defense for the infant, but inflammatory complications like mastitis may lead to the cessation of breastfeeding. Antisecretory factor (AF) has a role in the regulation of secretory processes and inflammation. The objective of the study was to describe AF-levels in plasma and breast milk, and in relation to breast complications. Breastfeeding mothers (n = 95) were consecutively recruited at a Well Baby Clinic in Umeå, Sweden. At inclusion four weeks postpartum, samples of venous blood (10 mL) and breast milk (10 mL) were collected. Active AF was analyzed with ELISA using a monoclonal antibody mAb43, and was detected in all samples of plasma and breast milk with a positive correlation (Spearman coefficient = 0.40, p < 0.001; Pearson correlation = 0.34, p < 0.01). High AF-levels in plasma correlated with high AF-levels in breast milk. The results suggest a co-regulation between active AF in plasma and breastmilk, and/or a local regulation of AF in the breast. Further studies are needed to determine the pathways for the activation of AF-levels in breast milk and plasma.

Experimental staphylococcal mastitis in bitches: clinical, bacteriological, cytological, haematological and pathological features.

The objectives of the work were to study the features of experimentally induced canine mastitis and to present hypotheses regarding the pathogenesis of the disease. The right caudal abdominal mammary gland of six bitches was inoculated on day 8 after whelping with Staphylococcus intermedius to induce mastitis; adjacent mammary glands were used as controls. Clinical examination, bacteriological and cytological (whiteside test, Giemsa) examination of mammary secretion, as well as haematological tests were performed from 5 days before until 34 days after challenge. Mastectomy was sequentially performed 1, 2, 4, 18, 26 and 34 days after challenge in each of the bitches, in order to carry out a pathological examination of mammary glands. All animals developed clinical mastitis: challenged glands became painful, hot, enlarged and oedematous; secretion was brownish, purulent, with flakes or clots, subsequently becoming yellowish and thick. Staphylococci were isolated from all inoculated glands (up to 22 days). WST was positive in 41/46 samples from inoculated glands and 66/138 samples from control glands; neutrophils predominated during the acute stage. Blood leukocyte counts increased, whilst platelet counts decreased. Gross pathological findings initially included congestion, purulent discharge and subcutaneous oedema; then abscesses, brownish areas and size decrease were seen. Salient histopathological features were initially neutrophilic infiltration, haemorrhages, destruction of mammary epithelial cells and alveoli, and then infiltration by lymphocytes, shrunken alveoli, loss of glandular architecture and fibrous tissue proliferation. We conclude that in bitches, intrammamary inoculation of Staphylococcus intermedius can induce clinical mastitis, followed by subclinical disease. The disorder is characterized by bacterial isolation and leukocyte influx in challenged glands, by leukocyte presence in adjacent mammary glands, by increased blood leukocyte counts and by destruction of mammary parenchyma.

Milk C-reactive protein in canine mastitis.

Presence of mastitis in lactating bitches can become life threatening for both the bitch and pups. The aim of the present study was to evaluate a possible utility of C-reactive protein (CRP) in both milk and serum for canine mastitis diagnosis. Our study showed that milk CRP levels ranged between 0.1 and 4.9µg/mL and from 0.3 to 40.0µg/mL in healthy and diseased bitches (P<0.01), respectively, while serum CRP levels ranged between 2.0 and 8.6µg/mL and between 0.3 and 162.3µg/mL in healthy and diseased bitches (P<0.01), respectively. Milk and serum CRP levels were higher in both clinical and subclinical mastitis when compared with healthy controls (P<0.05 in all cases). However, no significant differences were recorded in CRP concentrations between clinical and subclinical cases. Based on these results, it could be concluded that serum and milk CRP could be useful in order to diagnose canine mastitis.

Whole blood transcriptome analysis reveals potential competition in metabolic pathways between negative energy balance and response to inflammatory challenge.

Negative Energy Balance (NEB) is considered to increase susceptibility to mastitis. The objective of this study was to improve our understanding of the underlying mechanisms by comparing transcriptomic profiles following NEB and a concomitant mammary inflammation. Accordingly, we performed RNA-seq analysis of blood cells in energy-restricted ewes and control-diet ewes at four different time points before and after intra mammary challenge with phlogogenic ligands. Blood leucocytes responded to NEB by shutting down lipid-generating processes, including cholesterol and fatty acid synthesis, probably under transcriptional control of SREBF 1. Furthermore, fatty acid oxidation was activated and glucose oxidation and transport inhibited in response to energy restriction. Among the differentially expressed genes (DEGs) in response to energy restriction, 64 genes were also differential in response to the inflammatory challenge. Opposite response included the activation of cholesterol and fatty acid synthesis during the inflammatory challenge. Moreover, activation of glucose oxidation and transport coupled with the increase of plasma glucose concentration in response to the inflammatory stimuli suggested a preferential utilization of glucose as the energy source during this stress. Leucocyte metabolism therefore undergoes strong metabolic changes during an inflammatory challenge, which could be in competition with those induced by energy restriction.

[Variations of peripheral blood autoantibody, immunoglobuliln, and complement levels in patients with non-lactational mastitis and their clinical significances].

OBJECTIVE: To detect the variations in peripheral blood levels of autoantibodies, immunoglobulilns and complements in patients with non-lactational mastitis and investigate whether non-lactational mastitis is an autoimmune disease with immune dysfunction. METHODS: Seven-eight patients with non-lactational mastitis treated in our hospital between September 2013 and May 2015 and 88 healthy women (control) were examined for peripheral blood levels of antinuclear antibody (ANA), anti-histone antibody (AHA), immunoglobulins (IgA, IgM, and IgG) and complements (C3, C4, and total complements). RESULTS: s Of the 78 patients with non-lactational mastitis, 50 (64.10%) were positive of ANA showing mainly the granular and cytoplasmic granular fluorescence patterns, and the positivity rate was significantly higher than that in the control group (P<0.000). Twenty-eight (36.00%) of the patients were positive of AHA, a rate significantly higher than that in the control group (P<0.000). The levels of IgA, IgM, C4, and total complements levels were all significantly elevated in the patients compared with those in the control group (P<0.05). CONCLUSION: Patients with non-lactational mastitis have abnormal changes in peripheral blood levels of immunoglobulins and complements with high positivity rates for ANA and AHA, indicating that non-lactational mastitis is an autoimmune disease with immune dysfunction.

The course of acute-phase proteins and serum cortisol in mastitis metritis agalactia (MMA) of the sow and sow performance.

The objective of this study was to evaluate changes in acute-phase proteins (APPs) during mastitis metritis agalactia (MMA) in sows. Sows with MMA (group one, n=15) and healthy sows (group two, n = 15) were evaluated at days 0, 5, 10, 15, and 20 postpartum. Number of total born, liveborn, stillborn, and mummified pigs did not differ significantly between the groups. Preweaning mortality was higher (P < 0.001) among MMA sows than among healthy control animals. The offspring of healthy sows had higher (P < 0.05) weaning litter weights than the off-spring of MMA sows. Mean serum alpha 1-acid glycoprotein (AGP) concentrations were higher in MMA sows on the days 1 (P < 0.05), 5 (P = 0.05), and 10 (P < 0.001) post partum. Mean serum haptoglobin (HPT) was higher in MMA sows on days 1, 5 (P < 0.001), and 10 (P < 0.05) of lactation. Cortisol serum concentrations up to day 10 post partum were higher (P < 0.001) in MMA sows than in healthy sows. AGP was negatively correlated with litter weight, indicating that activation of the cellular immune response in sows negatively affects the growth rate of suckling piglets. Correlations were found between the overall means for weight, acute-phase proteins, and serum cortisol concentration.

Host responses associated with chronic staphylococcal mastitis in rabbits.

Staphylococcal infection causes substantial economic losses in commercial rabbit production systems, and is associated with a wide variety of lesions, including chronic suppurative mastitis, which mainly affects breeding females. Most chronic staphylococcal infections in rabbits are caused by the ST121 lineage of Staphylococcus aureus, although other less common lineages, such as ST96 can also be involved. The aims of the present study were to characterise the host immune response in natural cases of mastitis in rabbits caused by S. aureus, to evaluate any relationship between peripheral and local immunity and to investigate the effect of different S. aureus genotypes on these immune responses. Adult multiparous female rabbits that were affected with chronic staphylococcal mastitis (n = 204) were enrolled into the study. Histological and immunohistochemical evaluations of mammary glands were undertaken, as well as flow cytometric analyses of blood. S. aureus isolates from the mammary glands were identified by multilocus sequence typing. Differences in the number of infiltrating cells were detected, depending on the type of pathology, with more immature lesions demonstrating greater cellularity, characterised by greater numbers of T lymphocytes, macrophages and plasma cells. A relationship was seen between the cells in blood and mammary tissues, the most notable being the positive correlation between monocytes and tissue macrophages. When glands were infected with ST96 strains, fewer granulocytes (P < 0.01) and greater numbers of B cells (P < 0.01), T cells (P < 0.001), CD4(+) T cells (P < 0.001) and CD8(+) T cells (P < 0.01) were detected, compared with mammary glands that were infected by ST121 strains of S. aureus.

Transition diseases in grazing dairy cows are related to serum cholesterol and other analytes.

The objectives of this study were to describe the incidence of postpartum disease and to evaluate the association with serum cholesterol concentrations during the first 3 weeks after calving in grazing dairy cows. The association between non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BHBA), calcium and postpartum diseases was also evaluated. A total of 307 Holstein dairy cows from 6 commercial grazing herds in Osorno, Chile, were monitored from calving until 21 days in milk. Cases of retained placenta, clinical hypocalcemia and clinical mastitis were recorded by the farmer using established definitions. Twice weekly, cows were evaluated for metritis by the same veterinarian based on vaginal discharge and body temperature. Postpartum blood samples were collected weekly and analyzed for serum concentrations of cholesterol, NEFA, BHBA and calcium. Cows were considered as having subclinical ketosis if BHBA >1.2 mmol/L, and subclinical hypocalcemia if calcium <2.0 mmol/L in any of the 3 weekly samples. Overall, 56% of the cows studied developed at least one clinical or subclinical disease after calving. Incidence of individual diseases was 8.8% for retained placenta, 4.2% for clinical hypocalcemia, 11.7% for clinical mastitis, 41.1% for metritis, 19.9% for subclinical hypocalcemia and 16.6% for subclinical ketosis. Lower postpartum cholesterol in cows was associated with developing severe metritis or having more than one clinical disease after calving. For every 0.4 mmol/L decrease in serum cholesterol cows were nearly twice as likely to be diagnosed with multiple clinical diseases after calving. Higher BHBA concentrations and lower calcium concentrations during week 1 were associated with severe cases of metritis. Low serum calcium concentration during week 1 was also associated with developing more than one clinical disorder after calving. In conclusion, the incidence of postpartum diseases can be high even in grazing herds and lower serum cholesterol concentrations were associated with occurrence of clinical postpatum disorders.

Benign breast disease I: hormonal investigation.

One hundred eighty-four patients with benign breast disease (BBD) were studied and compared with 50 normal women. All of the women had ovulatory cycles according to a biphasic basal body temperature and a plasma prolactin in the normal range. Their corpus luteum function was evaluated by way of plasma progesterone (P) and estradiol (E2) determinations at days 5, 7, and 9 of the hyperthermic phase. In the 184 patients, plasma P over plasma E2 ratio during the luteal phase was found significantly lower than in normal women. When the patients were grouped according to type of breast lesions, it appeared that plasma P was constantly lower in all groups than in the normal women, while plasma E2 was either normal or elevated in the groups of patients with adenosis tumors and increased nodularity of both breasts. From these results it may be postulated that an imbalance in the secretion of E2 and P by the corpus luteum is a constant finding in women with benign breast disease.

Quantification of osteopontin in human plasma with an ELISA: basal levels in pre- and postmenopausal women.

OBJECTIVES: To develop an immunoassay for osteopontin (OPN), a secreted phosphoglycoprotein that is implicated in a number of human diseases, and establish basal plasma OPN levels in healthy women. DESIGN AND METHODS: An antigen-capture ELISA was developed to quantity OPN in plasma using a combination of mouse monoclonal and rabbit polyclonal antibodies. Basal OPN levels were determined in blood plasma of 21 pre- and 14 postmenopausal women obtained at 7-day intervals over a 4-week period. RESULTS: A group of 35 healthy women had a median OPN level of 31 micrograms/L (range = 14-64 micrograms/L). Comparison between pre- and postmenopausal women showed that their 4-week average OPN levels did not differ significantly (p > 0.16, Mann-Whitney test), and that levels in each premenopausal individual remained constant during the menstrual cycle, unaffected by cyclical levels of leuteinizing hormone and progesterone. CONCLUSION: Systematic quantification of plasma OPN can now be done by ELISA, which was used to establish basal plasma OPN levels in a group of healthy women. Levels in pre- and postmenopausal women appeared relatively stable over a 4-week period.