Flaxseed oil supplementation alters the expression of inflammatory-related genes in dogs.

Long chain n-3 fatty acids are beneficial to mammals because of their anti-inflammatory role. However, whether flaxseed oil, which is rich in short chain n-3 fatty acids, has such a role, it has not been extensively examined. This study investigated the supplementation of flaxseed oil on the regulation of genes involved in inflammatory responses such as heat shock proteins (HSP90 and HSP70) and interleukin (IL1ß) in the white blood cells of dogs. Five beagles and 5 greyhounds were supplemented with Melrose(®) flaxseed oil at the rate of 100 mL/kg food for 21 days. The blood was collected at day 0, 15, and 22 following supplementation. The expression of 3 genes was quantified using real-time polymerase chain reaction. Plasma concentrations of fatty acids such as alpha linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid, and arachidonic acid were measured, and their correlations with changes in gene expression were determined. Flaxseed oil supplementation downregulated the expression of HSP90 and IL1ßin greyhounds but showed no significant effect on these genes in beagles. HSP70 remained unchanged in both breeds following the supplementation. Correlations of HSP90 and IL1ßexpression levels with the plasma fatty acid concentrations on day 22 showed a significant negative correlation in greyhounds. Dietary flaxseed oil altered the expression of genes involved in inflammation in white blood cells. Because the expression of the genes may vary in different breeds, it will be useful to consider breed responses to dietary manipulation in canine nutrition management.

Comparison of testing saliva and serum for detection of antibody to human immunodeficiency virus in Jamaica, West Indies.

BACKGROUND: In Jamaica the reported incidence of AIDS increased from 0.1/100000 in 1985 to 20.2/100000 in 1995. Here there is great reluctance to have voluntary blood testing and, indeed, any blood testing. Since only enzyme-linked immunoassay (EIA) was available for screening serum HIV-1 and 2 antibody, it was considered that a non-invasive saliva screening EIA could be an advantageous alternative. OBJECTIVE: this study was designed to evaluate the OraScreen HIV Rapid Test, a new, simple saliva screening EIA for anti-HIV-1&2 and to compare its sensitivity and specificity with a standard serum anti-HIV screening EIA in current use in Jamaica. STUDY DESIGN: specificity and sensitivity of HIV antibody assays were compared in matched serum and saliva samples obtained from 257 volunteers from a family planning clinic and from visa applicants, representing a low risk population (Group I), and from 52 volunteers known to be HIV infected (Group II). RESULTS: in Group I, 257 volunteers of unknown HIV status, one was positive for anti-HIV-1 in both serum and saliva. One other was seropositive but negative on saliva testing; confirmatory Western Blot (WB) testing on this serum was negative and this subject was tabulated as blood HIV negative. Fifty-one of the known seropositive volunteers (Group II) were saliva antibody positive. One saliva sample was inadequate and this individual was excluded from the study. Serum samples from three others in Group II were grossly haemolysed but their saliva samples were antibody positive. CONCLUSION: With the exclusion of one subject whose saliva sample was inadequate, the OraScreen HIV Rapid Test showed 100% specificity identifying 256/256 HIV antibody negative individuals, and 100% sensitivity by identifying 52/52 infected individuals as HIV antibody positive.

Expression of polymorphic msp1beta genes during acute anaplasma Marginale rickettsemia.

Immunization of cattle with native MSP1 induces protection against Anaplasma marginale. The native immunogen is composed of a single MSP1a protein and multiple, undefined MSP1b polypeptides. In addition to the originally sequenced gene, designated msp1beta(F1), we identified three complete msp1beta genes in the Florida strain: msp1beta(F2), msp1beta(F3), and msp1beta(F4). Each of these polymorphic genes encodes a structurally unique MSP1b protein, and unique transcripts can be identified during acute A. marginale rickettsemia. The structural polymorphism is clustered in discrete variable regions, and each MSP1b protein results from a unique mosaic of five variable regions. Although each of the MSP1b proteins in the Florida strain contains epitopes recognized by serum antibody induced by protective immunization with the native MSP1 complex, the variable regions also include epitopes expressed by some but not all of the MSP1b proteins. These data support testing recombinant vaccines composed of the multiple antigenically and structurally unique MSP1b proteins combined with MSP1a in order to mimic the efficacy of native MSP1 immunization.

Brucellosis in adult beef cattle of Mexican origin shipped direct-to-slaughter into Texas.

OBJECTIVE: To compare prevalence estimates of brucellosis (BR) in adult beef cattle that originated from different states and regions of Mexico and that were shipped direct-to-slaughter into Texas during 1995. DESIGN: Epidemiologic survey. ANIMALS: About 65,000 adult beef cattle. PROCEDURE: Blood samples were collected during postmortem examinations and were tested for serum antibodies to Brucella abortus, using the particle concentration fluorescence immunoassay and automated complement-fixation test. Prevalence estimates and 95% confidence intervals of BR were calculated by state of origin in Mexico. The difference among prevalence estimates of BR in cattle from different states and regions was tested for significance (P < 0.05), using the proportion test. RESULTS: On the basis of serologic test results, the overall prevalence estimate of BR was 0.32%. The prevalence estimate of BR in cattle from the state of Chihuahua (0.10%) was significantly different than that in cattle from the states of Nuevo Leon (0.23%), Zacatecas (0.34%), Durango (0.47%), Chiapas (1.81%), Tamaulipas (2.71%), Aguascalientes (7.89%), and Campeche (12.24%). In addition, prevalence estimates of BR in cattle were significantly different among the northern (0.22%), south-central (3.18%), and south coastal (9.42%) regions of Mexico. CLINICAL IMPLICATIONS: Results of this study indicate that the number of cattle exposed to B abortus may be significantly different among states and regions of Mexico. Current import sanitary requirements should continue to mitigate potential risk of transmission of BR from sexually intact cattle of Mexican origin to Texas cattle.

Tuberculosis in adult beef cattle of Mexican origin shipped direct-to-slaughter into Texas.

OBJECTIVE: To evaluate differences in prevalence of tuberculosis (TB) in adult beef cattle that originated from different states in Mexico and were shipped direct-to-slaughter into Texas in 1995. DESIGN: Epidemiologic survey. ANIMALS: Approximately 65,000 adult beef cattle. PROCEDURES: Postmortem examinations of carcasses for detection of Mycobacterium bovis infection were conducted at slaughter plants in Texas. Specimens were collected from cattle with granulomatous lesions, stored in neutral-buffered 10% formalin or saturated sodium borate solution, and processed for histologic and bacteriologic diagnosis. Prevalence and 95% confidence intervals were estimated by state of origin. Difference between prevalences for different states was tested for significance (P < 0.05), using the proportion test. RESULTS: Overall prevalence of TB at slaughter in adult beef cattle that originated from Mexico was approximately 0.5/1,000 (34/65,233). Prevalence of TB in cattle that originated from Chihuahua (0.07) was significantly lower than that in cattle from Coahuila (0.80), Nuevo Leon (1.27), and Tamaulipas (1.81). CLINICAL IMPLICATIONS: Prevalence of M bovis infection in adult beef cattle may be significantly different between states in the northern border region of Mexico. On the basis of disease prevalence and numbers of exported cattle and provided safeguards such as TB testing are continued, cattle from Chihuahua may pose a lower risk of TB transmission to Texas cattle than do cattle from Coahuila, Nuevo Leon, and Tamaulipas. To allow interstate/international movement of cattle from northern border states of Mexico, TB testing requirements should be continued. In the context of international trade, southern border states of the United States should continue collaborating with northern border states of Mexico to control and eradicate this disease.

Love styles among Guatemalans in a local village.

81 participants in a village in Guatemala were administered a translated version of the 1986 Love Attitudes Scale of Hendrick and Hendrick. Analysis showed that the men scored significantly higher on the Ludus love style than the women. Research among other Latinos outside the United States is suggested.

Enhanced cardiac metabolism of plasma glucose in high-altitude natives: adaptation against chronic hypoxia.

The metabolism of glucose in mammalian heart is 25-50% more O2 efficient than the metabolism of free fatty acids. To assess the role of substrate preference in adaptations to chronic hypoxia, positron emission tomographic measurements of heart regional glucose uptake rates after an overnight fast were made in volunteer Quechua subjects and in Sherpa subjects, both indigenous to altitudes of over 3,000 m, and in a group of lowlander volunteers. Highest uptake rates were found in the Quechuas on arrival and in the Sherpas after a 3-wk period at low altitude, intermediate rates in Quechuas after a 3-wk period at low altitude and in the lowlanders, and lowest rates in Sherpas on arrival. These low values were probably related to the stress of travel to the site of the experiments. Measured plasma catecholamines, hormones, and substrates indicated that glucose concentrations correlated best with observed variations in glucose uptake, with a negative correlation for the control subjects and a positive correlation for the Quechuas and Sherpas. Uptake values in Quechuas declined significantly after a 3-wk period at low altitude, but the positive correlation with glucose levels persisted. We conclude that an elevated glucose preference in heart is a true metabolic adaptation in humans adapted over generations to chronic hypoxia.

Cross-reactivity between murine melanoma antigen B700 and a human melanoma-associated antigen (M-66) recognized by autologous antibody: evidence suggesting shared epitopes.

We have previously reported the purification and partial characterization of a human melanoma-associated antigen (M-66) recognized by autologous antibody. This antigen was found to be an unusually acidic 66 kDa glycoprotein. In studies of murine melanoma, a 67-kDa albumin-like melanoma-associated antigen (MAA) isolated from B16 melanoma cells has also been reported by our laboratories. Because the murine MAA, B700, has a molecular weight that is nearly the same as M-66, we sought to determine what similarities and differences existed between these two antigens. Human sera S150, which is known to recognize M-66, was found to bind to murine melanoma cell line B16. The addition of purified M-66 inhibited binding of S150 to B16 cells. Binding by S150 was not noted against murine melanoma cell line S91, which is known not to express cell surface B700. Conversely, reactivity of S150 against Y-Mel 84:420, known to express M-66, could be inhibited by preincubation with B16 cells. Four monoclonal antibodies known to recognize B700 were evaluated for-binding against murine B16 and human melanoma cell line Y-Mel 84:420. Binding was noted against both B16 and Y-Mel 84:420 which could be inhibited by the addition of M-66. Binding of S150 was also noted against purified B700 as tested by ELISA. While a comparison of the amino acid composition of the two antigens revealed similarities, M-66 contained 2.8 times as much serine and 0.4 times as much proline as B700. B700 has been reported to be related to serum albumin, which is not the case for M-66.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation and purification of a squamous cell carcinoma of the head and neck-associated antigen identified by autologous antibody.

We have previously shown that detection of autologous antibody activity to squamous cell carcinoma of the head and neck may be augmented by dissociation of immune complexes. Western blot analysis with autologous antibody has identified a 60 kDa squamous cell carcinoma of the head and neck-associated antigen in spent media and immune complex-dissociated serum ultrafiltrate not recognized by normal human sera. Antigen-containing fractions of spent media were eluted from anion exchange columns immediately after serum albumin indicating that the antigen has an acidic pI < 4. Preparative purification of the squamous cell carcinoma antigen was accomplished by anion exchange of concentrated spent media (protein concentration 300 mg/ml) followed by lectin affinity chromatography with a Triticum vulgaris column. A single 60 kDa band was detected by silver stain and Western blot in antigen-containing fractions eluted following lectin affinity chromatography and SDS-PAGE. Final concentration of the antigen was determined to be 1 microgram/ml of protein with relative activity increased 1600 x over unfractionated spent media. We conclude that a squamous cell carcinoma of the head and neck-associated antigen, detected by autologous antibody, is an acidic 60 kDa glycoprotein.

Purification and partial characterization of a shed 66 kDa melanoma-associated antigen identified by autologous antibody.

We have previously reported the isolation of a 66 kDa melanoma-associated antigen, identified by autologous antibody, in serum and unfractionated spent tissue culture media by Western blot analysis. The antigen, detected by autologous serum S150, was found to be broadly represented on melanoma, glioma, renal cell carcinoma, neuroblastoma and head and neck carcinoma cell lines. S150 did not react with bladder or colon carcinoma, fetal fibroblasts, pooled platelets, lymphocytes and red blood cells, autologous cultured lymphocytes or fetal calf serum. To further characterize the antigen, spent tissue culture media, obtained from autologous melanoma cell line, Y-Mel 84:420, was separated by an isoelectric focusing column. Unabsorbed control serum S150 was noted to have a maximum titer of 1:2040 against autologous melanoma cells as measured by protein A hemadsorption. Following isoelectric focusing the greatest decrease in autologous antibody titer (30-fold) occurred with fractions having a pI between 2 and 3. Further resolution of the antigen was accomplished with high-pressure ion-exchange chromatography. One of these fractions showed a significantly higher concentration of antigen and was distinctly resolved from bulk serum albumin. Subsequent Western blot analysis, with autologous antibody, of the isolated antigen-containing fraction, confirmed the presence of a single 66 kDa band. Exposure of the antigen, purified by high-pressure ion-exchange chromatography, to neuraminidase ablated recognition by autologous antibody and suggests that sialic acid is present on the protein and may be part of the antigenic epitope. Binding of antigen, obtained following DEAE anion exchange chromatography, was noted to lectins derived from Triticum vulgaris, Dolichos biflorus and Lycopersicon esculentum. Preparative purification of the antigen was accomplished by anion exchange followed by lectin affinity chromatography with a Dolichos biflorus column. Antigen obtained following lectin affinity chromatography subjected to SDS-PAGE and silver stain revealed a single band at 66 kDa. We conclude that a melanoma-associated antigen detected by autologous antibody in spent tissue culture media is an unusually acidic glycoprotein (pI 2-3).

Identification of a unique IgG Fc binding site in human intestinal epithelium.

In experiments to determine whether serum antibodies in patients with Crohn's disease could be used as probes for detecting potentially etiologic Ag in the patients' tissues, we found that peroxidase (HRP)-labeled IgG from healthy persons, as well as from the patients, bound to normal colonic and small intestinal epithelium, mostly or entirely to goblet cells. The binding was due to a reaction involving the Fc region of IgG because HRP-labeled Fc fragments of IgG bound, but HRP-Fab, HRP-IgA, and HRP-bovine albumin did not, and because binding of HRP-IgG was inhibited competitively by unlabeled IgG or Fc fragments but not by IgG Fab fragments or IgA. These immunohistochemical results were confirmed by ELISA with microtiter wells coated with a sonicated homogenate from human colonocytes. The epithelial IgG Fc binding site was characterized by SDS-PAGE as consisting of a high Mr (greater than 200,000 Da) and a 78,000-Da component. It bound all four subclasses of human IgG and bound aggregated as well as monomeric IgG. It is distinct from known human Fc-gamma R by lack of recognition by mAb to those receptors and differences in affinity for various subclasses of human and murine IgG. This unique IgG Fc binding site might be involved in immunologic defense of the gut, perhaps by mediating reactions between foreign Ag and the contents of goblet cells.

Pro-development soap operas: a novel approach to development communication.

PIP: Soap operas have their roots in 18th century English romance novels. These evolved into serialized radio dramas. In their current form, they were developed primarily to attract large audiences in order to sell consumer products. Hence the name soap which refers to the soap manufacturers who commonly advertise on such programs. In the world of soap operas there are 2 kinds. Those that function primarily to entertain and sell consumer products, and those that primarily entertain, but infuse positive social messages. The former are found everywhere, but are the only kind in America. The latter are found exclusively in developing countries. American soap operas have conveyed pro-social messages in the past, but they differ fundamentally from pro-development soap operas in their theoretical foundations. American soap operas are created by people who want to sell consumer goods. Development soap operas are created by people who want to convey pro-social messages that will aid their country's development. Both must be popular in order to be successful, but the former lack moral coherency, are unrealistic, erode values, and are created through a process of a theoretical development; while the latter have moral coherency, are realistic, promote values, and are created through a process of theoretical development. The 1st pro-development soap opera was Ven Conmigo (Come With Me) and was produced in Mexico between 1975-76. Its primary purpose was to increase adult literacy. During the year it ran, applicants at adult literacy centers rose by 600,000 or 63% compared to 7% the year before, and 2% the year after. The 2nd pro-development soap opera was Acompaname (Accompany Me) and it primary purpose was to promote family planning. It ran from 1977-78 and during that time the number of family planning adopters rose by 560,000 and contraceptive sale sin Mexico rose sharply. The question of what are pro-social messages and who should control them must be answered by each country in its effort to increase development.^ieng

Pulmonary lysozyme--a secretory protein of type II pneumocytes in the rat.

A 14 kDa basic protein isolated from rat lung lavage was demonstrated to be lysozyme by its amino acid sequence analysis. An antiserum to rat lysozyme stained type II pneumocytes and alveolar macrophages. In rat lungs, no staining of the airway cells was noted. Lysozyme was detectable in type II pneumocytes by immunocytochemistry and by a quantitative immunoassay of lung homogenates of fetal lungs at Day 20 of gestation. An increase in the lysozyme content of the lung with increasing gestational and postnatal age of the rat was noted. In adult animals, lysozyme accounts for about 169.0 micrograms/g of wet lung weight and 0.3% of the soluble proteins in lung homogenate. Lysozyme constitutes about 6.6% of the total soluble proteins in rat lung lavage. Metabolic labeling and immunoprecipitation were used to demonstrate that rat type II pneumocytes synthesize and secrete lysozyme in vitro. However, in human lungs, lysozyme was identified in serous submucosal glands but not in alveolar type II pneumocytes. The results demonstrate differential distribution of a secretory protein in rodent and human lungs and indicate that in the rat lysozyme could be used as an immunohistologic marker for type II pneumocytes and as an indicator of secretory activity and maturation of type II pneumocytes.

Lack of correlation between infection with reovirus 3 and extrahepatic biliary atresia or neonatal hepatitis.

Infection with reovirus 3 (Reo-3) has been suggested as the cause of extrahepatic biliary atresia and idiopathic neonatal hepatitis, but confirmation has been lacking. Therefore we have searched for a specific anti-Reo-3 antibody response in the sera of patients with biliary atresia or neonatal hepatitis and for Reo-3 antigens in their hepatobiliary tissues. Sera from 23 infants with extrahepatic biliary atresia, 12 with neonatal hepatitis, 30 age-matched control patients with other liver diseases, and 55 control patients without liver disease were tested by an enzyme-linked immunosorbent assay for total (IgA, IgG, and IgM) anti-Reo-3 antibodies; sera of infants younger than 6 months of age were tested also for IgM anti-Reo-3 antibodies alone. There was no difference between either total or IgM anti-Reo-3 antibody levels in infants with extrahepatic biliary atresia or neonatal hepatitis and levels in control infants. Reo-3 antigens were not detected in the hepatobiliary tissues of 19 infants (18 with biliary atresia, one with neonatal hepatitis) by an immunoperoxidase method that readily demonstrated Reo-3 in control infected HEp-G2 cells. Our data do not support a relationship between neonatal liver diseases and infection with Reo-3.

Influence of genital herpes on results of fluorescent treponemal antibody absorption test

Both the fluorescent treponemal antibody absorption (FTA-ABS) test and Venereal Disease Laboratory (VDRL).

The hemodynamic effects of intrauterine hypoxia: an experimental model in newborn lambs.

An experimental animal model of intrauterine hypoxia and respiratory distress in newborn lambs was produced by inducing maternal hypotension. Serial hemodynamic data indicated that the oxygenation defect in the lambs was due to right-to-left shunting of blood through fetal channels rather than within the lungs. Shunting was mainly across the foramen ovale, but, in severely distressed animals, significant right-to-left shunt also occurred through the ductus arteriosus. Left-to-right shunts across the ductus arteriosus were found in lambs with milder respiratory distress. The implications of perinatal hypoxia as it affects the pulmonary vascular bed in human neonates with the respiratory distress syndrome (hyaline membrane disease) and persistence of the fetal circulation are discussed. It is speculated that the early pulmonary vascular esponses in the two diseases may be identical.

Inflammatory myopathy, IgA deficiency, and intestinal malabsorption.

Two IgA-deficient children with inflammatory myopathy and intestinal malabsorption were evaluated. The myopathy was characterized by weakness of facial and proximal limb muscles, increased serum concentrations of lactic dehydrogenase and creatine phosphokinase, and histologic evidence of inflammation and degeneration of muscle fibers. Features of the intestinal abnormality were blunted villi, interstitial inflammation, and reduction in IgA-containing plasma cells and IgA content of epithelial cells. The myopathy and malabsorption improved with corticosteroid treatment. Circulating antibodies to striated muscle could not be demonstrated in either patient, but one had antibodies to milk and chicken serum proteins. We speculate that IgA deficiency may predispose to the development of inflammatory myopathy.