Behavioral effects of acute sublethal exposure to dimethoate on wood mice, Apodemus sylvaticus: II--Field studies on radio-tagged mice in a cereal ecosystem

The effects of direct exposure to an organophosphate (OP) pesticide, dimethoate, were examined in free-living wood mice, Apodemus sylvaticus, in a wheat field. Male mice were radio-tagged at night and followed during 2-3-d periods, before and after an intraperitoneal injection of 50 mg/kg dimethoate which previous laboratory studies had demonstrated causes a maximum depression in brain acetylcholinesterase (AChE) activity of 75% relative to non-exposed mice. In subsequent weeks, survival was estimated by tracking and trapping data. Exposure to dimethoate significantly decreased locomotor activity in the first 6 h after administration resulting in a significant decrease in the area over which animals moved. These effects were limited to the night of treatment and disappeared 24 h later. The transient behavioral impairment of the dimethoate-treated animals appeared to have no effect on medium-term survival. Direct exposure to OPs sufficient to cause 75% depression of brain AChE is unlikely to be hazardous to wood mice if exposure is transient, as in the present study; it would be expected that sustained exposure sufficient to cause such behavioral effects would, however, be detrimental.     

Schizorchis arfaai sp. n. (Cestoda: Anoplocephalidae) from the wood-mouse, Apodemus sylvaticus, in Caspian area of Iran

An artificial, slowly expanding mass which compressed the spinal cord produced histological, electrophysiological, and neurological changes. A comparison was made of the effects of laminectomy in the cat with dorsally and ventrally placed masses. Laminectomy reversed declining cord function and produced prompt recovery of somatosensory evoked potentials in cats with dorsally placed masses. In contrast, a worsening of these functions occurred following laminectomy in animals with ventrally placed masses. In all cases, evoked potential changes were consistent with neurological status and were predictive of the outcome of surgery.     

Apparent digestibility of nutrients in diets with different energy density, as estimated by direct and marker methods for pigs with or without ileo-cecal cannulas

The objective of this study were a) to compare the apparent total tract digestibility (TD) between non-cannulated (intact) and cannulated (steered ileo-cecal valve technique, SICV) pigs fed diets differing in energy density (Exp. 1) and b) to compare the direct vs marker (Cr2O3) methods for estimation of the TD and apparent ileal digestibility (ID) in SICV-cannulated pigs (Exp. 2). In Exp. 1, 24 intact and 18 SICV-cannulated castrates of approximately 40 kg initial BW were randomly assigned to six treatments in a 2 x 3 x 2 factorial arrangement (two pig types, three carbohydrate sources, and two fat levels). In Exp. 2, the same SICV-cannulated pigs from Exp. 1 were given those treatments in a 2 x 3 x 2 factorial arrangement (two methods of digestibility estimation, three carbohydrates sources, and two fat levels). In both experiments either cornstarch, soybean hulls, or pure cellulose, without or with fat, were incorporated into a barely-soybean meal based diet to alter energy density. Daily diets were isoenergetic (based on NEf), and water supply was .33 L/MJ of NEf. In Exp. 1, the pig type effect on the TD of DM, OM, CP, and the pig type x carbohydrate interactions for the TD of DM, OM, and crude fiber (CF) were significant (P < .05), merely due to a larger difference found for the diet enriched with cellulose. In Exp. 2, the TD and ID evaluated with the marker method were significantly lower (except for the TD of CF) than with the direct method, mainly because Cr recovery was below 100%. Overall, the marker method seems to be superior because the TD means obtained from Cr ratios were closer to the TD obtained from intact pigs. In general, the SICV technique seems to be suitable for long-term digestibility studies to measure the TD and ID in the same pig fed low-or high-fiber diets.     

Sequencing of the mtDNA cytochrome b gene and reconstruction of the matriarchal relationships between wood and field mice of the genus Apodemus (Muridae, Rodentia)

The primary sequence of a 402-bp part of the cytochrome b gene was determined in nine species of wood and field mice of the genus Apodemus. The majority of mutations were synonymous. The total number of transitions exceeded than of transversions. Among all substitutions, C-T transitions prevailed (51%); the most common substitution type in genus-specific sites was C-A transversions (42%). In interpopulation analysis, only transitions were recorded. A phylogenetic tree, constructed with the use of the neighbor-joining method, showed that the genus Apodemus is divided into three highly divergent groups: south Asian (Apodemus argenteus, A. semotus), east Asian (A. speciosus, A. agrarius), and Eurocaucasian (A. sylvaticus, A. flavicollis, A. uralensis, A. ponticus, A. flavipectis). The mean genetic distances within each group were 12.6, 11.2, and 8.8%, respectively. The species of the first group are more remote genetically and ancestrally with regard to the other groups. The interspecies divergence estimated for A. speciosus ranged from 0.25 to 3.75%. Thus, the evolutionary age of the genus Apodemus is about 6 Myr, and time of divergence of A. speciosus populations is 0.1-1.5 Myr. The phylogeny inferred from the data on the sequence of the cytochrome b gene in Apodemus mtDNA is somewhat different from similar phylogenies based on other genetic data and from the zoological taxonomy of wood and field mice. However, the above classification of species is confirmed by features of their karyotypes and segmentation of satDNA, and by the RFLP of total nDNA and isozyme polymorphism. Our results are in good agreement with the new classification of wood and field mice recently proposed by Russian zoologists.     

Total serum IgE values in two different populations of asthmatics with different levels of environmental contamination

A group of asthmatic patients with an average age between 15 to 50 years were followed at the Allergology Service of Teaching Hospitals from Santa Clara and Sagua la Grande, respectively. For the diagnosis, clinical and espirometric criteria were taken into account; the sample consisted of 100 patients living in Sagua; this part is considered to be a very polluted area, and the same amount of patients from Santa Clara City. The seric figures of total IgE were quantified and a survey was applied to value its relationship with other polluting substances and other clinical and epidemiological aspects of bronchial asthma. The number of patients from Sagua with high levels of IgE was relevant, this could be influenced by the high levels of polluting substances which act as inhaler or irritant substances.     

Chromosome variation in two populations of the crested newt, Triturus cristatus carnifex, with different melanoma incidence

Morphometric analysis of the chromosomes of the crested newt, Triturus cristatus carnifex, was carried out for two populations, one from Rosate (Milan) and one from Bagnaia (Perugia), which differed in the prevalence of melanoma (2.9 and 0.8%, respectively). Chromosomes were arranged in pairs both by the cytologist and by a previously developed computer programme. The two populations had significant differences in the relative lengths of chromosomes 9 and 10, and in the arm ratio values of 6 and 11. Though the morphometric parameters of the other elements of the complement appeared to be similar, the Rosate population showed significantly higher chromosomal asymmetry. In the Rosate population, there was greater intra-population and intra-individual variability than in the Bagnaia population. The karyotype variations observed suggest that chromosomal rearrangement may have occurred.     

Identification of two distinct CD5- B cell subsets from human tonsils with different responses to CD40 monoclonal antibody

This study investigated the response of different CD5- B cell subsets to CD40 monoclonal antibody (mAb) in various combinations with interleukin (IL)-4 or rabbit anti-human mu chain antibody (a-mu-Ab). The different CD5- B cell subsets were isolated from tonsillar B cell suspensions depleted of CD5+ B cells and subsequently fractionated on Percoll density gradients. While resting CD5+ B cells proliferated and produced IgM molecules in response to a-mu-Ab, IL-4 and CD40 mAb as well as to Staphylococcus aureus Cowan strain I (SAC) and IL-2, resting CD5- B cells, which were co-purified in the same 60% Percoll fractions, consistently failed to respond. These cells were, however, activated by the stimuli employed, as demonstrated by their capacity to express the surface activation markers CD69, CD25 and CD71. Resting CD5+ B cells had the typical phenotype of mantle zone B cells (IgM+ IgD+ CD39+ CD38- CD10- CDw75dim), whereas resting CD5- B cells were CD38- CD39- CD10- CDw75 intermediate and expressed surface IgM but relatively little surface IgD and could not be classified as mantle zone or germinal center cells. The finding that purified germinal center cells (CD38+ CD10+ CD39- CDw75bright, IgG+) responded to CD40 mAb and IL-4 and also to SAC plus IL-2 further underlined the differences to resting CD5- B cells. However, some of the data collected suggest possible relationships between CD5- B cells and germinal center cells. The CD5- B cells isolated from the 50% Percoll fraction proliferated in response to a-mu-Ab, CD40 mAb and IL-4 as well as to SAC and IL-2. These cells had the same mantle zone B cell phenotype as the CD5+ B cells, but their capacity to respond to the stimuli in vitro was unrelated to a possible contamination with CD5+ B cells, as documented by the appropriate controls. Furthermore, upon exposure to SAC or phorbol esters, the large majority of CD5- B cells from the 50% Percoll fraction did not express surface CD5 and there was very little if any accumulation of CD5 mRNA. Finally, most of the cycling cells in the stimulated CD5- B cells did not express CD5. The CD5- B cells from the 50% Percoll fraction were comprised of a consistent proportion of cells that expressed surface activation markers.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cloning of total mRNA populations from adult and embryonic mice

Total clone banks of cDNAs synthesized from poly(A)-RNA obtained from three stages of the developing mouse were constructed. The stages chosen were 13-day-old embryo, neonatal, and fully grown adult. To have as complete a bank as possible, large numbers of individual clones were generated approximately 400,000 for the 13th day embryo and neonatal mouse and approximately 610,000 for the adult bank. In each case the clone bank was constructed by inserting double stranded cDNA into the PstI site of pBR322 by the "G-C tailing" method. Sequences cloned in this way could be separated from the plasmid host DNA by treatment of the resultant total chimeric plasmid population with PstI. Aliquots of the cloned cDNA material were labeled with 32P by "nick translation" using Escherichia coli DNA polymerase I for the preparation of hybridization probes. Back-hybridization of these probes to the total clone banks allowed the determination of the sequence diversity among the above three very different developmental stages. The use of such clone banks should allow the identification of developmental stage specific mRNAs.     

Cholesterol oxidation in meat from chickens fed alpha-tocopherol- and beta-carotene-supplemented diets with different unsaturation grades

The production of B-ring and side-chain oxysterols was evaluated in meat from chickens fed diets differing by the kind of oil or fat added. The effect of supplementary levels of natural antioxidants, as alpha-tocopherol and beta-carotene, on the meat cholesterol oxidative stability was also studied. Lard, sunflower and olive oil were used as dietary fat. Raw and cooked meats were analyzed for oxysterols, and cholesterol was also quantified. Oxysterol analyses were carried out by combining the use of solid-phase extraction, thin-layer chromatography, capillary gas chromatography, and capillary gas chromatography-mass spectrometry. Oxysterols were detected within the 0.1-0.5 microg/g range in raw meat. Cooking increased the oxysterol content of the meat, and levels as high as 5 microg/g muscle tissue were observed. B-Ring oxysterols were mainly produced: the alpha- and the beta-epoxycholesterols, the 7alpha- and 7beta-hydroxycholesterols, and the 7-ketocholesterol. The results showed that the meat from the chickens fed the olive oil-based diet containing alpha-tocopherol at 200 mg/kg of diet presented the best cholesterol oxidative stability. A positive effect could not be found for dietary beta-carotene administered at levels of 15 and 50 mg/kg of diet. Furthermore, a significant decrease in the tissue cholesterol content was observed with the olive and the sunflower oil-based diets.     

Comparison of the effects of two low fat diets with different alpha-linolenic:linoleic acid ratios on coagulation and fibrinolysis

The formation of germinal centers (GCs) represents a crucial step in the humoral immune response. Recent studies using gene-targeted mice have revealed that the cytokines tumor necrosis factor (TNF), lymphotoxin (LT) alpha, and LTbeta, as well as their receptors TNF receptor p55 (TNFRp55) and LTbetaR play essential roles in the development of GCs. To establish in which cell types expression of LTbetaR, LTbeta, and TNF is required for GC formation, LTbetaR-/-, LTbeta-/-, TNF-/-, B cell-deficient (BCR-/-), and wild-type mice were used to generate reciprocal or mixed bone marrow (BM) chimeric mice. GCs, herein defined as peanut agglutinin-binding (PNA+) clusters of centroblasts/centrocytes in association with follicular dendritic cell (FDC) networks, were not detectable in LTbetaR-/- hosts after transfer of wild-type BM. In contrast, the GC reaction was restored in LTbeta-/- hosts reconstituted with either wild-type or LTbetaR-/- BM. In BCR-/- recipients reconstituted with compound LTbeta-/-/BCR-/- or TNF-/-/BCR-/- BM grafts, PNA+ cell clusters formed in splenic follicles, but associated FDC networks were strongly reduced or absent. Thus, development of splenic FDC networks depends on expression of LTbeta and TNF by B lymphocytes and LTbetaR by radioresistant stromal cells.     

Percutaneous absorption of ibuprofen from different formulations. Comparative study with gel, hydrophilic ointment and emulsion cream

In a three-way cross-over study on 6 healthy adult volunteers, the percutaneous absorption of ibuprofen (CAS 15687-27-1) was studied with 3 topical formulations containing 5% w/w ibuprofen in a gel (Iprogel) or in a hydrophilic ointment or in an emulsion cream. By analysis of the plasma drug concentrations appearing after topical application, the relative drug bioavailability was calculated in terms of Cmax (maximum blood concentration of the drug), AUC (area under the curve of drug plasma concentrations at various time points) and Tmax (the time required for appearance of maximum drug concentration in the blood). The gel formulation showed the highest drug concentration in blood, reached in the shortest period, whereas that from the hydrophilic ointment showed the lowest drug concentration, reached at the slowest rate. The absorption from the reference product containing the drug as an o/w emulsion cream was less than with the gel formulation but higher than that found with the hydrophilic ointment.     

Molecular cloning, characterization and overexpression of two distinct cysteine protease cDNAs from Leishmania donovani chagasi

We have cloned and characterized two distinct cysteine protease cDNAs from Leishmania donovani chagasi. One of the cDNAs, Ldccy2, was isolated from a cDNA library prepared from total promastigote RNA while the other cDNA, Ldccys1, was isolated from a cDNA library prepared from total amastigote RNA. Ldccys2 has an open reading frame of 471 amino acids and Ldccys1 has an open reading frame of 447 amino acids. Comparison of the predicted protein sequences of the two distinct cysteine proteases with those of cysteine proteases from Leishmania pifanoi, a member of the L. mexicana complex, showed that the cysteine proteases from the two species of Leishmania are similar in their protein sequences. Each of the two cDNAs is distinct in genomic arrangement and chromosome location. Ldccys1 belongs to a family of cysteine proteases encoded by tandemly organized genes located on chromosome 7 while Ldccys2 appears to be a single cysteine protease gene located on chromosome 10. The organization of the two families of cysteine protease genes in L. donovani donovani was also found to be similar. In this species, the Lddcys1 genes are located on chromosome 5 while the Lddcys2 gene is located on chromosome 8. The Ldccys1 genes are expressed abundantly in the amastigotes recovered from infected hamsters, but at a very low level in the promastigote stage of development. On the other hand, the Ldccys2 gene is expressed both in the promastigote and amastigote stages. We have overexpressed the two cDNAs of cysteine proteases in Leishmania cells and the over-produced cysteine proteases are biologically active and are inhibited by cysteine protease inhibitors. Furthermore, the over-produced and indigenous amastigote specific cysteine protease, Ldccys1, reacted with polyclonal antibodies raised against this protein.     

Glucose and maltodextrin in enteral diets have different effects on jejunal absorption of nutrients, sodium and water and on flow rate in mini pigs

BACKGROUND: The effect of altitude on subjects between 8 to 22 and 23 to 51 years, that resided at sea level, or in the highlands, and developed altitude lung edema, is presented in series of 21 prospective registered patients at 4,800 to 5,000 m altitude in the Andes mountains of Ecuador. PATIENTS MATERIAL AND METHODS: The 21 sick patients were evaluated at 2 h and at 8 hr after exposition to the high altitude. They were immediately evacuated from the high altitudes. Clinical and radiological methods were used. The statistical analysis was performed with chi(2) test with Yates correction and relative risk. RESULTS: The younger patients tended to develop illness with a higher frequency (p < 0.01). The place of residence influenced the development of the illness only for the group of persons between the ages of 23 to 51 years (p < 0.03); and also in comparing all of the subjects from the coast with those from the highlands. CONCLUSIONS: A inverse relationship was found between the age and the possibility of the development of altitude pulmonary edema, possibly due t a basic incrementation of the pressure of the pulmonary artery, or because of a relative immaturity of the adaptative mechanisms. The place of residence was significantly associated with a higher frequency of pulmonary edema in persons between the ages of 23 to 51 years.     

Effect of monensin on milk production and efficiency of dairy cows fed two diets differing in forage to concentrate ratios

Four primiparous Holstein cows were gradually introduced, according to a Latin square design, to four diets obtained from the factorial combination of two forage to concentrate ratios (70:30 and 50:50) and two concentrations of monensin sodium (0 and 300 mg/d per cow). Addition of monensin tended to depress feed intake and milk fat content without affecting milk production and without interactions with forage to concentrate ratios. Ruminal propionate percentage was increased more by the addition of monensin to the low forage diet than by the addition of monensin to the high forage diet. Serum urea and concentrations of nonesterified fatty acids tended to decrease when monensin was added to the high forage diet but did not change when monensin was added to the low forage diet. The results suggested that monensin had moderate positive effects on efficiency of milk production and might have an antiketogenic effect with high forage diets.     

Growth performance, intestinal microbial populations, and serum cholesterol of broilers fed diets containing Lactobacillus cultures

A study was conducted to determine the effects of adherent Lactobacillus culture on growth performance, intestinal microbial population, and serum cholesterol level of broilers. Four dietary treatments, consisting of the basal diet (control), basal diet + 0.05, 0.10, or 0.15% Lactobacillus culture (LC), were fed to 2,000 Arbor Acres broiler chicks from 1 to 42 d of age (DOA). The chicks were randomly assigned to 40 cages (50 chicks per cage, 10 cages per diet). The experimental period was 42 d. Body weights and feed to gain ratio were measured at 21 and 42 DOA. The intestinal microbial populations and serum cholesterol levels were determined at 10, 20, 30, and 40 DOA. The results showed that body weights and feed to gain ratios were improved significantly (P < 0.05) when compared to control broilers for broilers fed diets containing 0.05 or 0.10% LC, but not 0.15% LC, at 21 and 42 DOA. Coliform counts in the cecum of birds receiving 0.05% LC at 10, 20, and 30 DOA, and 0.10% at 10 and 20 DOA were significantly lower (P < 0.05) than those of the control birds. The total aerobes, total anaerobes, lactobacilli, and streptococci in the small intestines and ceca of the control birds were not significantly different from those of the treated groups. Serum cholesterol levels were significantly lower (P < 0.05) in broilers fed the three diets containing LC at 30 DOA, and in the birds fed 0.05 or 0.10% LC at 20 DOA.     

Pneumocystis carinii organisms obtained from rats, ferrets, and mice are antigenically different

Pneumocystis carinii infections were developed in animals immunosuppressed by dexamethasone treatment either from activation of latent infection (ferret) or by transtracheal inoculation with P. carinii-infected lung tissue from the homologous species (rat or mouse). Convalescent-phase antisera were obtained by stopping dexamethasone treatment after 2 to 4 weeks and allowing animals 5 to 8 weeks for recovery. P. carinii harvests from infected lungs were purified by differential filtration, solubilized in buffer containing urea, sodium dodecyl sulfate (SDS), and 2-mercaptoethanol, subjected to SDS-polyacrylamide gel electrophoresis, and blotted to polyvinylidene difluoride sheets for Western immunoblot analysis. These lung preparations are hereafter referred to as P. carinii antigens. Convalescent-phase antisera from each animal species were reacted on Western blots of P. carinii antigens prepared from organisms isolated from rats, ferrets, or mice. Each combination of P. carinii antigens and antisera from the same species of animal reacted with three or more P. carinii antigen proteins. Convalescent-phase mouse antisera reacted with P. carinii antigens from mice but not rats or ferrets. Convalescent-phase rat antisera reacted with P. carinii antigens from rats and mice but not ferrets, and convalescent-phase ferret antisera showed reactions with ferret and mouse P. carinii antigens but not rat antigens. These findings indicate antigenic differences among P. carinii strains infecting these animals.     

Results and efficiency of programmed ventricular stimulation with four extrastimuli compared with one, two, and three extrastimuli

BACKGROUND: Conventional programmed ventricular stimulation protocols are inefficient compared with more recently proposed protocols. The purpose of the present study was to determine if additional efficiency could be derived from a 6-step programmed ventricular stimulation protocol that exclusively uses four extrastimuli. METHODS AND RESULTS: The subjects were 209 consecutive patients with coronary artery disease and documented sustained monomorphic ventricular tachycardia, nonsustained ventricular tachycardia, aborted sudden death, or syncope. These patients underwent 159 electrophysiological tests in the absence of antiarrhythmic drug therapy and 105 electrophysiological tests in the presence of antiarrhythmic therapy. Programmed stimulation was performed with two protocols in random order in each patient. Both protocols used an eight-beat drive train, 4-s intertrain pause, and basic drive cycle lengths of 350, 400, and 600 ms. The 6-step protocol started with coupling intervals of 290, 280, 270, and 260 ms, which were shortened simultaneously in 10-ms steps until S2 was refractory. The 18-step protocol used one, two and three extrastimuli in conventional sequential fashion. The end points were 30 s of sustained monomorphic ventricular tachycardia, two episodes of polymorphic ventricular tachycardia requiring cardioversion, or completion of the protocol at two right ventricular sites. There was no significant difference in the yield of sustained monomorphic ventricular tachycardia using the two protocols, regardless of the clinical presentation or treatment with antiarrhythmic drugs. Polymorphic ventricular tachycardia occurred with the 18-step protocol twice as frequently as with the 6-step protocol (6% versus 3%, P < .001). The duration of the 18-step protocol was significantly longer than that of the 6-step protocol in patients with inducible ventricular tachycardia (5.5 +/- 7 versus 2.3 +/- 2 minutes, P < .001), as well as in patients without inducible ventricular tachycardia (25.4 +/- 7 versus 6.9 +/- 2 minutes, P < .001). CONCLUSION: A stimulation protocol that exclusively uses four extrastimuli improves the specificity and efficiency of programmed ventricular stimulation without compromising the yield of monomorphic ventricular tachycardia in patients with coronary artery disease.     

Familial Mediterranean fever and hyperimmunoglobulinemia D syndrome: two diseases with distinct clinical, serologic, and genetic features

OBJECTIVE: To determine whether the 2 periodic febrile syndromes familial Mediterranean fever (FMF) and hyperimmunoglobulinemia D syndrome (HIDS) are distinct diseases. METHODS: Clinical manifestations of the diseases were analyzed by physicians experienced with FMF and HIDS. Serum immunoglobulin (Ig) levels were studied in 70 patients with FMF using nephelometry or ELISA and compared with Ig levels in 50 patients with HIDS. Genetic linkage of HIDS with the chromosome 16 polymorphic locus RT70, currently used for refined localization of the FMF susceptibility gene (MEFV), was studied in 9 HIDS families (18 patients) using polymerase chain reaction amplification and gel electrophoresis. RESULTS: The main clinical features distinguishing FMF from HIDS were lymphadenectomy, skin eruption, and symmetrical oligoarthritis in HIDS, and monoarthritis, peritonitis, and pleuritis in FMF. Increased IgG levels were found in 12 patients with FMF (17%), IgA in 16 (23%), IgM in 9 (13%), and IgD in 9 (13%), significantly lower than the prevalence reported for HIDS. We found no evidence for genetic linkage between HIDS and the chromosome 16 marker RT70. CONCLUSION: HIDS and FMF are different entities, clinically, immunologically, and genetically.