Biological effects of anti-RANKL antibody administration in pregnant mice and their newborns.

Denosumab, a fully human monoclonal antibody that neutralizes receptor activator of nuclear factor-κB ligand (RANKL) and blocks osteoclast differentiation, has received approval in Japan for use as an anti-resorptive drug for osteoporosis and skeletal-related events (SREs) in patients with solid cancer. Denosumab is contraindicated during pregnancy, though the effects of blocking RANKL activity on pregnant mothers and their newborns are unclear. We used mice to investigate the effects of an anti-RANKL antibody on maternal and newborn health. Mothers injected with the anti-RANKL antibody had increased bone mass as compared with the controls, while osteoclast number and the level of tartrate-resistant acid phosphatase (TRAP) in serum were increased at the end of pregnancy. Newborn mice exposed to the antibody in utero were normally born, but showed increased bone mass and died within 48 h after birth. None of the newborns were found to have milk in their stomachs, suggesting that they died due to a maternal defect in lactation. Consistent with this, anti-RANKL antibody-injected mothers displayed impaired mammary gland development. However, fostering by healthy surrogate mothers rescued only 33% of the antibody-exposed newborns, suggesting that neonatal mortality was due, at least in part, to an intrinsic defect in the newborns. Our findings show that anti-RANKL antibody administration during pregnancy results in not only an undesirable increase in bone mass, but also has harmful effects on newborn survival.

Synthesis of Recombinant P48 of Mycoplasma agalactiae by Site Directed Mutagenesis and its Immunological Characterization.

Contagious agalactia caused by Mycoplasma agalactiae is an economically important disease of sheep and goats and has been prevalent worldwide including India. The present study was undertaken to evaluate the membrane protein P48 of M. agalactiae for specific diagnosis of disease. For this, p48 gene of the organism was amplified by PCR and subjected to site directed mutagenesis to convert three TGA codons to TGG's and, subsequently, cloned into prokaryotic expression vector pPRO EX HTb. Purified recombinant P48 protein reacted to anti-P48 serum in western blotting, which confirmed its immunogenic nature. Furthermore, the immune-blotting of the cell lysates from various Indian isolates of M. agalactiae against anti-P48 serum resulted in a single band at âˆ¼ 48 kDa among all isolates, indicating the conserved nature of P48 antigen in M. agalactiae. Also, the cross reactivity of P48 antigen among various Mycoplasma spp. was checked by western blotting which revealed reactivity only with M. agalactiae and M. bovis. Hence, this antigen could be exploited to differentiate M. agalactiae from other pathogenic Mycoplasma species except M. bovis. However, the inability of P48 to distinguish M. agalactiae from M. bovis does not downgrade the significance of P48 as the two species are usually host specific.

Late primary autoimmune hypothyroidism in a patient with postdelivery autoimmune hypopituitarism associated with antibodies to growth hormone and prolactin-secreting cells.

BACKGROUND: Pituitary and thyroid autoimmunity can be triggered by pregnancy. We report the first association of combined growth hormone (GH) and prolactin secretion deficiency due to autoimmune damage to GH- and prolactin-secreting cells in a patient with postdelivery lactation failure, presenting subsequently with primary autoimmune hypothyroidism. PATIENT FINDINGS: A 34-year-old woman presented with lactation failure following the delivery of her first child. She had a family history of hypothyroidism without a history of pituitary dysfunction. Physical examination did not show any abnormal findings. Laboratory investigations showed normal gonadotropin levels after the restoration of normal menstrual cycles following pregnancy, normal basal and stimulated cortisol levels, but an impaired GH response to insulin-induced hypoglycemia, and low basal prolactin and insulin-like growth factor-1 concentrations. Thyroid function was normal when initially investigated three months after delivery, but five months later, marked primary hypothyroidism (thyrotropin levels >100 mIU/L) occurred. Immunological investigation revealed the presence of antipituitary antibodies, identified by double immunofluorescence and targeting GH- and prolactin-secreting cells. Antithyroid antibodies, in the normal range three months postpartum, became significantly elevated when the hypothyroidism appeared. Autoimmune hypophysitis is responsible for selective or multiple pituitary-hormone deficiencies, sometimes involving thyrotropin secretion and causing secondary hypothyroidism, but usually associated with hyperprolactinemia. To our knowledge, this is the first observation of autoimmune hypopituitarism involving deficient growth hormone and prolactin secretion in a patient with lactation failure after delivery, subsequently followed by severe primary autoimmune hypothyroidism, thus falling into an unusual constellation of autoimmune polyendocrine syndrome type 3. CONCLUSIONS: Considering the well-known relationship between pregnancy and autoimmunity, an early postdelivery immunological and functional investigation in women presenting with disorders of lactation may be useful to detect potential pituitary and thyroid dysfunction even at a subclinical stage.

[The role of staphylococci in the occurrence, development and chronicity of lactation mastitis]. / Rol' stafilokokkov v vozniknovenii, razvitii i khronizatsii laktatsionnykh mastitov.

The investigation results obtained by the authors and the data of literature on the role of the biological properties of staphylococci in the pathogenesis, development and chronization of acute suppurative lactation mastitis are generalized. The concept of lactation mastitis as a monoetiological disease caused solely by bacteria of the genus Staphylococcus is substantiated. The importance of the individual properties of staphylococci at different stages of the pathogenesis of lactation mastitis, as well as their role in ensuring the resistance of the infective agent to the protective mechanisms of the body and in the chronization of the suppurative process, is considered. The data on the use of a number of biological properties of staphylococci as markers for the prognostication of the course of lactation mastitis are presented and their prognosticating effect at different tactics of surgical treatment is evaluated.

[The information value of the biological properties of the causative agent in the prognosis of the duration of the course of suppurative inflammatory diseases of staphylococcal etiology]. / Informativnost' biologicheskikh svoistv vozbuditelia pri prognozirovanii dlitel'nosti techeniia gnoino-vospalitel'nykh zabolevanii stafilokokkovoi étiologii.

The informative value of biological characteristics of Staphylococcus aureus reflecting the ability of microbes to inactivate certain factors of natural resistance (lysozyme, complement, immunoglobulins, bactericidal component of interferon) was established in determining the duration of the caused by them pyo-inflammatory diseases such as postinfection abscesses, lactation mastitis. On this basis the heterogeneous successive procedure of pattern recognition was used to develop algorithms allowing prognosis of the duration of pyo-inflammatory diseases of staphylococcal etiology in 84.6-94.4% of the patients after the quantitative and qualitative assessment of the biological properties of the pathogen with not less than 90% reliability.

Breastfeeding anaphylaxis case study.

This case describes a woman who experienced an anaphylatic reaction associated with breastfeeding. The reaction occurred with each feeding on day three postpartum and resolved on day four. Possible reasons for this severe reaction are suggested.

Suppression of interferon response of bovine leukocytes during clinical and subclinical ketosis in lactating cows.

The influence of spontaneous ketosis on interferon alpha and gamma production in blood leucocytes and on PHA induced lymphocyte blastogenic response was investigated. Twenty three cows 4.13 +/- 2.8 weeks after calving were divided into three experimental groups on the basis of blood ketone bodies, glucose and free fatty acids concentrations. The leukocytes of cows with clinical symptoms and the highest concentration of ketones and free fatty acids in blood responded with the lowest levels of interferons alpha and gamma to three interferon inducers: Newcastle Disease Virus (NDV), phytohemagglutinin (PHA) and concanavalin A (ConA). Depression in interferon PHA stimulated synthesis correlated with a very low mitogenic response of blood lymphocytes. Blood leukocytes of cows with subclinical ketosis, characterized by mild clinical symptoms and a lower concentration of ketones in blood in comparison to cows with clinical ketosis, responded better to interferon and mitogenic stimulation; however, the interferon titer and blastogenesis were still lower than in leukocytes of healthy cows. Correlation between the stage of ketosis and the level of interferon production in milk leukocytes was also observed. A possible relationship between the suppression of interferon production in blood leukocytes and the increased concentration of ketone bodies in blood is discussed.

Effects of cold treatment and ketosis induced by starvation on interferon production in leukocytes of lactating cows.

The production of interferons in blood and milk leukocytes of three groups of cows was measured to determine the effect of 6-days cold treatment (-2 degrees to -8 degrees C) and/or starving. The first group (cold) was treated with low ambient temperature (-2 degrees C to -8 degrees C) 11 hours every day for 6 days, the second (cold and starved) was treated with low temperature and starved for 6 days. The third group (controls) was fed normally and kept in a barn at room temperature (18 degrees to 20 degrees C). The leukocytes of the control and the cold treated cows responded normally to interferon induction with Newcastle Disease Virus (NDV) and mitogens: phytohemagglutinin (PHA) and concanavalin A (ConA). The cows treated with low temperature and starved for 6 days developed biochemical blood changes of ketosis. Leukocytes of these cows with ketosis produced less interferon (p less than 0.05) than before starvation and less than leukocytes of the control cows and the cold treated cows. It can be assumed that ketosis caused by starving decreases the ability of a cow's leukocytes to produce interferons.

Interactions of heat stress and bovine somatotropin affecting physiology and immunology of lactating cows.

During summer, 34 cows received daily injections of placebo or 25 mg of bST and were placed in a thermoregulated or a heat stress environment. Heat stress increased rectal temperatures, respiration rates, and plasma cortisol concentrations and decreased milk yield. Four of 9 bST-treated cows and none of 8 control cows became atactic on the 1st d of heat stress. When exposed to heat stress, cows treated with bST experienced higher rectal temperatures throughout the trials than cows treated with placebo. Nonetheless, bST increased milk yields in both environments. The major effect of heat stress on immune function was decreased migration of leukocytes to the mammary gland after chemotactic challenge. This effect of heat stress was not altered by bST. In summary, hyperthermia induced by heat stress and associated changes were greater for cows treated with bST. Detected effects of heat stress on the immune system were few and were not alleviated by bST. Use of bST during summer in subtropical climate zones requires careful management to avoid overexposure of bST-treated cows to heat stress.

Depression of polymorphonuclear leukocyte function associated with experimentally induced Escherichia coli mastitis in sows.

In a study of susceptibilities of sows from 2 herds to experimentally induced Escherichia coli mastitis, a marked difference was seen. The "susceptible" sows were from a conventional herd and "resistant" sows were from a specific-pathogen-free herd. The purpose of the study was to determine whether deficient neutrophil function was associated with increased susceptibility to E coli-induced mastitis. Four in vitro procedures were used to evaluate polymorphonuclear leukocyte (PMN) function: (i) random migration under agarose, (ii) ingestion of 125I-iododeoxyuridine-labeled Staphylococcus aureus, (iii) quantitative nitroblue tetrazolium reduction, and (iv) iodination. After parturition and intramammary inoculation with E coli, sows from the susceptible herd were neutropenic and the neutrophils which were present in the peripheral blood had reduced function. Specifically, there were depressed random migration under agarose, S aureus ingestion, and iodination when compared with PMN function in resistant sows. These data indicate that susceptibility to E coli mastitis was associated with deficiencies in PMN numbers and function. Potential causes of the neutrophil dysfunction are discussed and include possible systemic hormonal aberrations or the presence of an inapparent viral or bacterial infection.

[Immunological reactivity of patients undergoing combined treatment of acute suppurative lactation mastitis]. / Immunologicheskaia reaktivnost' bol'nykh pri kompleksnom lechenii ostrogo gnoinogo laktatsionnogo mastita.

The analysis of treatment of 769 patients with acute mastitis enabled the authors to recommend the method of active surgical treatment which allowed shortening the time of the hospital and following ambulant treatment of the patients. The immune reactivity was found to be decreased in patients with severe and continuous course of the disease. So, the complex therapy of such patients must include immunological drugs.

Distribution of antibodies to rotavirus in serum and lacteal secretions of naturally infected swine and their suckling pigs.

Rotavirus antibodies were demonstrated in lacteal secretions and sera of 20 parturient sows and in sera of their newborn by an enzyme-linked immunosorbent assay blocking technique, using bovine rotavirus cell culture antigen and monospecific antibody to bovine rotavirus. Antibodies to rotavirus occur in the 3 immunoglobulin (Ig) classes IgM, IgA, and IgG in lacteal secretions. High and long-persisting antibody activity was mainly associated with the IgA class. The IgM and IgG decreased to undetectable concentrations in most sows during the 14-day investigation period. Serum antibodies of newborn pigs nursing their dams also decreased rapidly during this time. The heterologous enzyme-linked immunosorbent assay blocking technique was a reliable and rapid procedure for the demonstration of rotavirus antibodies.

[Use of heterologous antistaphylococcal gamma-globulin in treating suppurative mastitis]. / Primenenie geterologichnogo protivostafilokokkovogo gamma-globulina pri lechenii gnoinogo mastita.

The effectiveness of heterologic antistaphylococcal gammaglobulin was studied in the clinic in 67 cases of acute lactogenous mastitis. The activity of the preparation was 200 A. E/ml. The Bezredko method of injection was used--l ml once a day during 5 days. The treatment resulted in the decrease of intoxication, purification of the surgical wound, normalization on non-specific protection factors, rise of antistaphylococcal antitoxin titers and decrease of autosensitization phenomena.