Extracorporeal shockwave therapy raises mechanical nociceptive threshold in horses with thoracolumbar pain.

BACKGROUND: Although extracorporeal shockwave therapy (ESWT) is a common treatment for horses with back pain, effects on mechanical nociceptive threshold (MNT) and multifidus muscle cross sectional area (CSA) in the spine are unknown. OBJECTIVES: To evaluate effects of ESWT on spinal MNT and multifidus muscle CSA in horses with thoracolumbar pain. STUDY DESIGN: Non-randomised trial. METHODS: Thoracolumbar spines of 12 horses with thoracolumbar pain were radiographed to document existing pathology. Each horse received three ESWT treatments, 2 weeks apart (days 0, 14, 28). Palpation scores were documented (days 0, 45 and 65). Ultrasonographic CSA of left and right multifidus muscles was recorded at T12, T14, T16, T18, L3 and L5 (days 0, 45 and 65). MNT was measured at the same spinal sites every 7 days (day 0-56). RESULTS: Mechanical nociceptive threshold in 10/12 horses (83%) was greater at each time point compared with day 0 (P < 0.05). Mechanical nociceptive threshold increased at all time points at six sites in 2/12 (16%), at five sites in 3/12 (25%), at four sites in 4/12 (33%) and at one site in 1/12 (8%; P < 0.05). Mechanical nociceptive threshold average per cent  increase from day 0 to 56 was 64% for the thoracic region (T12-T18) and 29% for the lumbar region (L3-L5). There was no statistical difference in MNT from day 35 to 56 (P = 0.25). A bimodal analgesic trend was observed following ESWT. Degree of radiographic change was not associated with response to treatment. No significant change in multifidus muscle CSA was observed. MAIN LIMITATIONS: Small study size and lack of control group. CONCLUSIONS: Three treatments of ESWT 2 weeks apart raised MNT over a 56-day period in horses with back pain, but did not influence change in CSA of the multifidus muscle. While ESWT appears justifiable for analgesia, physiotherapeutic techniques may be necessary in conjunction for concurrent muscle rehabilitation.

Immunogenicity of Potomac horse fever vaccine when simultaneously co-administered with rabies vaccine in a multivalent vaccine or as two monovalent vaccines at separate sites.

BACKGROUND: Potomac horse fever (PHF) is a potentially fatal enterocolitis of horses caused by Neorickettsia risticii. The disease was originally recognised almost 40 years ago in the state of Maryland in the US. It is now known to occur in many areas of North America, as well as having been described in South America and Europe. Monocomponent PHF vaccines are available, but clinical protection with vaccination has been reported to be inconsistent. OBJECTIVES: This study was designed to assess the immunogenicity of a commercially available Potomac Horse Fever (PHF) vaccine when administered as either a monovalent PHF vaccine simultaneously co-administered with a separate monovalent Rabies vaccine or as a multivalent PHF/Rabies vaccine in horses. STUDY DESIGN: Randomised parallel group trial. METHODS: Ninety-one client or University owned horses participated in this open-label randomised study, with 45 horses receiving the monovalent vaccines at separate sites and 46 receiving the multivalent vaccine at a single site. Serum PHF IFA titres were determined twice prior to vaccination and at 1, 2 and 3 months after vaccination. RESULTS: Both vaccination protocols exhibited poor immunogenicity, with only one-third of all the animals demonstrating seroconversion, defined as an increase in titre of greater than 400 over baseline, at any time point after vaccination. The monovalent PHF vaccine exhibited significantly greater immunogenicity in terms of the number of horses exhibiting seroconversion, as compared to the multivalent vaccine, at one (20 vs. 11, P = 0.03) and two (18 vs. 9, p = 0.02) months post vaccination. The monovalent PHF vaccine also exhibited significantly greater immunogenicity in terms of the median (interquartile range) IFA titres, as compared to the multivalent vaccine, at one (800 [200-1600] vs. 400 [200-800], P = 0.009) and 2 months (400 [200-1600] vs. 400 [100-800], P = 0.02) post vaccination. There was no significant difference between groups at 3 months in either seroconversion rate or median IFA titers. MAIN LIMITATIONS: This study did not assess the actual protective effects of PHF vaccination but rather used the serologic response to vaccination as a surrogate biomarker of immunity. CONCLUSIONS: The multivalent PHF/Rabies vaccine exhibited lower immunogenicity as compared to the monovalent PHF vaccine co-administered with a separate Rabies vaccine.

Determination of cross-linking residues in a pharmaceutical polymer by liquid chromatography-high resolution full scan mass spectrometry.

A liquid chromatography-mass spectrometry (LC-MS) method was developed as limit test for an amine cross-linking residue in a pharmaceutical polymer. The method was based on full scan data with extracted ions for the accurate masses of dicyclohexylmethane-4, 4'-diamine (DMDA) and the internal standard 1,12-diaminododecane (DADD) obtained by Fourier transform MS. Dicyclohexylmethane-4,4-diisocyanate (DMDI) the reactive form of the cross-linking residue was determined as it decomposition product DMDA. Calibration curves for quantification of DMDA were linear in the range 2-100 ng/ml, the LOD was 1 ng/ml or 10 pg on column. Precisions/recoveries for spiked samples at the level of the limit of 1 ppm for DMDA and DMDI were +/-9.6%/38.6% and +/-14.5/10.0% (n=3), respectively. Unpredictable recovery was found in the extraction of polymer samples because of the complexity of the matrix and the reactivity of dicyclohexylmethane-4,4-diisocyanate (DMDI). PEG residues extracted from the polymer were found to cause ionization suppression and also affected the chromatography, these effects were reduced by using a gradient program. By using this method the level of amine residues in samples from different batches of polymers were determined to be much lower than the limit of 1 ppm. The method allowed comparison of the results obtained from the polymer before and after purification indicating that the residual DMDA could be decreased by a washing procedure.

Reduced glutathione and S-acetylglutathione as selective apoptosis-inducing agents in cancer therapy.

The effect of reduced glutathione (GSH) and S-acetylglutathione (S-acglu) treatment on several tumor cell lines and normal cells in vitro was investigated. GSH and S-acglu applied at concentrations of 1 mM and 2 mM induced apoptosis in malignant cells as shown by DNA-fragmentation and staining of apoptotic cells with 7-amino-actinomycin D while viability and growth of normal cells were not significantly influenced by this treatment. The results demonstrated that GSH and S-acglu may be selective inducers of apoptosis in malignant cells.

Generation of human monoclonal anti-idiotypic antibodies with specificity to the murine monoclonal anti-CA 125 antibody B43.13.

Two human monoclonal antibodies, HID-7E7 and ROB-6F2, were produced by EBV transformation of peripheral blood lymphocytes (PBL). PBL were obtained from a patient with ovarian cancer who had been exposed several times to a Tc-99m labeled murine monoclonal anti-CA 125 antibody (B43.13, Biomira, Edmonton) for immunoscintigraphy. The HID-7E7 and ROB-6F2 producing B-cells were cloned with a limiting dilution technique and have shown stable immunoglobulin secretion within a period of three years. The human monoclonal antibodies HID-7E7 and ROB-6F2 are of the IgG isotype, and bind with significant affinity to the murine monoclonal antibody B43.13, which was used for immunoscintigraphy. Binding affinity of ROB-6F2 to other murine antibodies could not be detected. Cross reactivity of HID-7E7 to a murine anti-CEA monoclonal antibody was observed. In order to verify the anti-idiotypic character of the generated human antibodies, the ability of HID-7E7 and ROB-6F2, respectively, to inhibit the formation of the CA125/B43.13 complex is demonstrated via an enzyme-linked immunosorbent assay. These human anti-idiotypic antibodies are possible candidates for immunotherapy of ovarian cancer in patients with a small tumor burden following surgery and/or chemotherapy.

Immunological profile of patients with ovarian-cancer under immunostimulation with murine monoclonal-antibodies.

The longer survival of ovarian cancer patients after immunostimulation has been connected with the induction of an anti-tumor activity triggered by cellular and humoral immune responses. Our interest was to study the long-term influence on the immune system in relation to the various levels of the HAMA response and the disease stage. The immunological profile was evaluated by regularly performing phenotyping and functional analysis of peripheral blood lymphocytes (PBL). We report the statistical analysis of immunological data obtained in a study with 77 ovarian cancer patients examined over a period of up to 28 months. The results demonstrate that these immunological data are important for monitoring cancer patients under immunotherapy whereas they provide no prognostic significance.

Interactions between tumor cells and tumor infiltrating lymphocytes in human ovarian carcinoma.

The aim of our study was to investigate the lymphocytic infiltration rate of ovarian tumors and the possible use of tumor-infiltrating lymphocytes (TIL) as a therapeutic tool in gynecologic oncology. Twelve tumors were treated with digesting enzymes in order to isolate tumor-infiltrating lymphocytes as well as tumor cells, TIL were expanded by culture in the presence of human interleukin-2 (IL-2). Freshly prepared tumor cells were allowed to grow in culture medium for several days before the first investigations were performed. TIL could only be isolated from 50% of the investigated tumors. In contrast to this the isolated tumor cells could be largely expanded in 73% of the cases. The expression of the CA125 antigen in the culture supernatants served as control and could still be evaluated up to three months after isolation. In parallel the antigen expression on the cellular surface was estimated by immunocytochemistry. Evaluating the phenotypes of TIL showed predominantly CD3+, their expansion rate was only poor. Tumor cells were isolated and expanded in order to test the tumor-directed cytotoxic efficacy of TIL and for further use in transplantation to nude mice.

Relevance of immunological parameters in progression of colorectal-carcinoma.

The long-term influence on the immunological stage from surgery and/or adjuvant or palliative therapy of 23 patients with metastatic colorectal carcinoma was investigated by performing regular phenotyping and functional analysis of peripheral blood lymphocytes (PBL). The following groups were chosen: A (n=6); patients after resection of primary tumor and liver-metastases without chemotherapy. B (n=3); patients with catheter implantation after resection of primary tumor and liver metastases receiving an adjuvant arterial chemotherapy with 5-fluorouracil (5-FU) and folinic acid (5-formyltetrahydrofolic acid, FA). C (n=7); patients with non-resectable liver-metastases, receiving arterial or systemic chemotherapy after catheter implantation. D (n=7); patients with extrahepatic filiae receiving systemic palliative chemotherapy. Lymphocytes of 10 healthy volunteers served as controls. Furthermore, we were able to show effects of 5-FU and FA on the immune system.

Improved enrichment of functionally intact anterior pituitary cells by sequential centrifugal elutriation and density gradient sedimentation.

By centrifugal elutriation and subsequent Percoll density gradient centrifugation, we obtained highly enriched cell populations of the anterior pituitary of ovariectomized adult rats. Enrichment of somatotrophs, gonadotrophs and mammotrophs was 90%, 80% and 77%, respectively (determined by immunocytochemistry). 41% of the GH-cells showed a medium cell size and the highest density. 52% of the LH-cells and 44% of the FSH-cells were found within the population of the large and medium-dense cells. 69% of the prolactin-cells belonged to the small-sized cells with low density. Thyrotrophs were found within all of the size ranges. The majority of the corticotrophs (82%) showed a small cell size. The enriched somatotrophs, LH-gonadotrophs, and mammotrophs responded to GRF, LRH and TRH. Before and after enrichment, somatotrophs and mammotrophs showed the same responsiveness to releasing hormones. In contrast, enriched LH-gonadotrophs exhibited a higher responsiveness to LRH than unfractionated LH-gonadotrophs.

Functional integrity of anterior pituitary cells separated by a density gradient.

Using a continuous Percoll density gradient, endocrine cells of the anterior pituitary were separated. The cells were obtained from adult female Sprague-Dawley rats which had been ovariectomized for 7 days. The gradient revealed two equally sized populations of cells with densities of about 1.02 and 1.09 g/ml. Ninety-two per cent of the cellular GH content, 64% of LH, and 60% of TSH were found in the high density peak. Sixty-one per cent of the cellular Prl appeared in the low density peak. Immunocytochemical staining of the LH containing cells showed that 74% of the gonadotrophs were in the high density peak. After separation, the cells retained their responsiveness to LRH, TRH and GRF. Culture conditions influenced stimulated hormone release. Before stimulation, the cells were cultured either in tissue culture flasks (attached cells) or in Petri dishes (cells in suspension) for 3 days. After TRH-stimulation, suspended thyrotrophs released more TSH than attached thyrotrophs. Comparing the cells of both peaks, attached thyrotrophs of the high density peak showed higher stimulated TSH-release than those of the low density peak. The response of the gonadotrophs and somatotrophs to stimulation did not differ when culture conditions were changed. The present results demonstrate that the secretory activity of endocrine cells is influenced by culture conditions and should be evaluated fore each cell type.

Migration and remittances: urban income and rural households in the Philippines.

PIP: Concerned about rural-urban ties and the role of remittances in the context of migration to a provincial city in the Philippines, it is argued that remittances must be examined within their social and cultural context and as part of a broader set of 2-way flows of people, goods, and money. The ways in whic remittances are used and their importance for persons in the rural areas need to be considered at the household level, and not only at the level of individuals or of villages. Both survey and case data were used based on research in Dagupan City, a regional urban center in the northern part of the Central Luzon Plain. The available evidence suggests that there is considerable migration into Dagupan City, but because there also is considerable out-migration, the net migration rate is low. Migrants to Dagupan maintain extensive ties with family elsewhere, and particularly with members of their natal families living at home in the rural areas. People, goods, and money move back and forth between city and rural home as migrants visit relatives and receive return visits from them. The flows and exchanges that occur may be divided into several categories for analytical purposes, although they often occur simultaneously: visiting; bringing material goods on visits; and sending material goods. Of the 176 migrants surveyed, only 7 were neither visiting nor sending something to relatives elsewhere. 92% of the migrants reported that they visit relatives outside Dagupan, while 69.9% received return visits from home. In remitting money, there was practically no difference between males and females in this regard. Age seemed to have some effect on the giving of money, with over 60% of those in the 26-35 age bracket reporting remittances, in contrast to only 38% of those in the 36-45 age group and 37% of those over 46 years old. There was little difference with respect to the maintenance of other ties among the different age categories. Length of time in Dagupan did not affect seriously the maintenance of ties. Overall 88 migrants, or 50%, remitted money. Overall, the percentage of income remitted varied considerably. There are no clearcut patterns indicating the types of ties likely to be maintained or the likelihood of remittances being given. 4 case studies are presented of individual migrants in their family context in order to provide a context for understanding both variation in amounts given and variation in the ways in which remittances are used.^ieng

Family strategies and the migration of women: migrants to Dagupan City, Philippines.

PIP: This study of women migrants to Dagupan, a 2ndry city in the Philippines, finds complex occupational and migration patterns with women in informal sector occupations as well as in professional/clerical ones. A large number of those in professional or clerical positions, who are also better educated, had moved away from home previously to study, whereas for a majority of those in the informal sector, including salesworkers, the move to Dagupan was the 1st in their life. While a majority of the former had moved with a definite job offer, the latter usually had come to Dagupan at the suggestion of relatives to look for work. Decision to move, usually made by the father, occurs within a cultural context which upholds strong expectations regarding the obligations of family members to 1 another, in this case, in the form of remittances. Daughters are expected to support families to a greater extent than sons, though both are encouraged to migrate to cities. 3 types of family strategies emerge in the analysis of migration pattersns: 1) rural households, usually poor and with little education, send their daughters out in the hope that remittances would help attain the basic necessities of life; 2) in an attempt to follow a strategy leading to upward mobility, some rural families educate their daughters, especially for occupations in the formal sector, whose remittances are used to educate other siblings; and 3) for those families with higher levels of education and income, encouraging daughters to migrate is done with a view to enhancing their position in society, rather than for the purpose of remittance. Women are encouraged by their families to migrate to cities with the expectation, based on strong cultural values, that such migration would help maintain the family as a unit through, among other things, financial support recieved.^ieng

Enzyme induction in Streptomyces hydrogenans. Comparison of the effects of different steroids to increase the activity of 3 alpha, 20 beta-hydroxysteroid dehydrogenase and of 3 beta, 17 beta-hydroxysteroid dehydrogenase.

After cultivation of Streptomyces hydrogenans in the presence of different steroids the activity of both 3 alpha, 20 beta-hydroxysteroid dehydrogenase and 3 beta, 17 beta-hydroxysteroid dehydrogenase was determined in the cell homogenate of the microorganism. By comparing the efficacy of the steroids to increase enzyme activities, steroids could be divided into 3 groups: a) steroids which stimulated preferentially the activity of 3 alpha, 20 beta-hydroxysteroid dehydrogenase (e. g., corticosterone), b) steroids which stimulated preferentially 3 beta, 17 beta-hydroxysteroid dehydrogenase (estradiol-17 beta), and c) those behaving intermediately (e. g., progesterone, 5 alpha -dihydrotestosterone). Highest 3 beta, 17 beta-hydroxysteroid dehydrogenase activity could be measured 2 h after addition of 5 alpha-dihydrotestosterone to the culture medium. The activity of 3 alpha, 20 beta-hydroxysteroid dehydrogenase, however, increased continuously up to 4 h. 3 alpha, 20 beta-hydroxysteroid dehydrogenase and 3 beta, 17 beta-hydroxysteroid dehydrogenase syntheses seemed to be controlled by steroids in a non-coordinate manner.

Purine nucleoside triphosphates in Streptomyces hydrogenans. Influence of steroids on the relative nucleotide level.

Various steroids in the culture medium of Streptomyces hydrogenans cause a rapid decrease of the relative GTP content of the cells. The relative ATP level is significantly diminished in the presence of progesterone and testosterone acetate. The initial decrease of the relative GTP content is proportional to the growth inhibitory effect of the steroids tested. There is no relationship between steroid-dependent induction of the synthesis of 20 beta- and 17 beta-hydroxysteroid dehydrogenase and the ability of the steroids to decrease the relative GTP content of the cells.

Immunological detection of 20beta-hydroxysteroid dehydrogenase-synthesizing polysomes from Streptomyces hydrogenans.

Immunoprecipitation of polyribosomes from Streptomyces hydrogenans synthesizing 20beta-hydroxysteroid dehydrogenase was performed by immunoelectrophoresis. Only polyribosomes from cells induced by 11beta, 21-dihydroxy-4, 11(20)-pregnadien-3-one in vivo were precipitated by immunoglobulins specific for 20beta-hydroxysteroid dehydrogenase. No unspecific precipitations with polysomes from control cells cultivated in the absence of steroids were observed. Therefore, 11beta, 21-dihydroxy-4, 17(20)-pregnadien-3-one increases the number of biological active polyribosomes involved in 20beta-hydroxysteroid dehydrogenase synthesis, presumably by specific stimulation of mRNA synthesis in Streptomyces hydrogenans.