Influence of environmental factors on the tropical peatlands diazotrophic communities from the Southern Brazilian Atlantic Rain Forest.

The tropical peatlands of southern Brazil are essential for the maintenance of the Atlantic Rain Forest, one of the 25 hotspots of biodiversity in the world. Although diazotrophic micro-organisms are essential for the maintenance of this nitrogen limited ecosystem, so far studies have focused only on micro-organisms involved in the carbon cycle. In this work, peat samples were collected from three tropical peatland regions during dry and rainy seasons and their chemical and microbial characteristics were evaluated. Our results showed that the structure of the diazotrophic communities in the Brazilian tropical peatlands differs in the evaluated seasons. The abundance of the genus Bradyrhizobium showed to be affected by rainfall and peat pH. Despite the shifts of the nitrogen-fixing population in the tropical peatland caused by seasonality it showed to be constantly dominated by α-Proteobacteria followed by Cyanobacteria. In addition, more than 50% of nifH gene sequences have not been classified, indicating the necessity for more studies in tropical peatland, since the reduction of N supply in the peatlands stimulates the recalcitrant organic matter decomposition performed by peatland micro-organisms, influencing the C stock.

Analysis of adenovirus DNA detected in rodent species from the Democratic Republic of the Congo indicates potentially novel adenovirus types.

Different species of adenoviruses (AdVs) infect humans and animals and are known for their role as pathogens, especially in humans, with animals, primarily rodents, often serving as model systems. However, although we know over 100 types of human AdVs, we know comparatively little about the diversity of animal AdVs. Due to the fact that rodents are the most diverse family of mammals and a standard model system for human disease, we set out to sample African rodents native to the Democratic Republic of the Congo and test them for AdV DNA using a semi-nested consensus PCR. A total of 775 animals were tested, and viral DNA was detected in four of them. The AdV DNA found belongs to three different AdVs, all being closely related to murine adenovirus 2 (MAdV-2). Considering the genetic differences of the amplicon were 9%, 11% and 19% from MAdV-2 and at least 10% from each other, they seem to belong to up to three different novel types within the Murine mastadenovirus B species. This evidence of genetic diversity highlights the opportunities to isolate and study additional AdVs that infect rodents as models for AdV biology and pathology.

Cholecystokinin antibody injected in cerebral ventricles stimulates feeding in sheep.

The role of brain cholecystokinin peptides in satiety was further assessed by using antibody to cholecystokinin to reduce cholecystokinin activity in the cerebrospinal fluid of sheep. Food intakes were increased approximately 100 percent during the 2-hour continuous injection of antibody into the cerebrospinal fluid. This supports the hypothesis that, during feeding, cholecystokinin is released into the cerebrospinal fluid, which transports it to the receptors that elicit satiety.

Brain cholecystokinin and nutritional status in rats and mice.

Under certain conditions, exogenously administered cholecystokinin (CCK) or its COOH-terminal octapeptide can terminate feeding and cause behavioral satiety in animals. Furthermore, high concentrations of CCK are normally found in the brains of vertebrate species. It has thus been hypothesized that brain CCK plays a role in the control of appetite. To explore this possibility, a COOH-terminal radioimmunoassay was used to measure concentrations of CCK in the cerebral cortex, hypothalamus, and brain stem of rats and mice after a variety of nutritional manipulations. CCK, mainly in the form of its COOH-terminal octapeptide, was found to appear in rat brain shortly before birth and to increase rapidly in cortex and brain stem throughout the first 5 wk of life. Severe early undernutrition had no effect on the normal pattern of CCK development in rat brain. Adult rats deprived of food for up to 72 h and rats made hyperphagic with highly palatable diets showed no alterations in brain CCK concentrations or distribution of molecular forms of CCK as determined by Sephadex gel filtration of brain extracts. Normal CCK concentrations were also found in the brains of four strains of genetically obese rodents and in the brains of six animals made hyperphagic and obese by surgical or chemical lesioning of the ventromedial hypothalamus. It is concluded that despite extreme variations in the nutritional status of rats and mice, CCK concentrations in major structures of the brain are maintained with remarkable constancy.

Bunker probe: A plasma potential probe almost insensitive to its orientation with the magnetic field.

Due to their ability to suppress a large part of the electron current and thus measuring directly the plasma potential, ion sensitive probes have begun to be widely tested and used in fusion devices. For these probes to work, almost perfect alignment with the total magnetic field is necessary. This condition cannot always be fulfilled due to the curvature of magnetic fields, complex magnetic structure, or magnetic field reconnection. In this perspective, we have developed a plasma potential probe (named Bunker probe) based on the principle of the ion sensitive probe but almost insensitive to its orientation with the total magnetic field. Therefore it can be used to measure the plasma potential inside fusion devices, especially in regions with complex magnetic field topology. Experimental results are presented and compared with Ball-Pen probe measurements taken under identical conditions. We have observed that the floating potential of the Bunker probe is indeed little affected by its orientation with the magnetic field for angles ranging from 90° to 30°, in contrast to the Ball-Pen probe whose floating potential decreases towards that of a Langmuir probe if not properly aligned with the magnetic field.

Characterization of the unusual insulin of Psammomys obesus, a rodent with nutrition-induced NIDDM-like syndrome.

Psammomys obesus fed a high-calorie diet develops a NIDDM-like syndrome. The use of reverse-phase high-performance liquid chromatography (HPLC) to study Psammomys insulin biosynthesis and release revealed a very delayed elution time for the Psammomys insulin peak appearing near the position of human proinsulin. This unusual peak was initially thought to represent partially processed insulin on the basis of its molecular size and susceptibility to trimming by carboxypeptidase B (CpB). However, the findings of an active carboxypeptidase E (CpE) enzyme and the normal amidated forms of gastrin and cholecystokinin octapeptide (CCK-8) in Psammomys tissues were inconsistent with CpE-related aberrant processing of insulin. Moreover, amino acid sequencing of the delayed peak of Psammomys insulin revealed fully processed insulin with amino acid sequence as predicted by the cDNA. The unique presence of a B-30 phenylalanine residue, resulting in an increased hydrophobicity of the insulin molecule, probably underlies the marked delay in elution time on HPLC. The unusual structure of Psammomys insulin does not appear to contribute to the proinsulinemia observed in diabetic Psammomys since the HPLC-purified molecule did not inhibit PC1 and PC2 convertase activities in an in vitro assay.

Expression of the cholecystokinin gene by cultured human primitive neuroepithelioma cell lines.

In this report we identify two cultured human primitive neuroepithelioma cell lines that express cholecystokinin (CCK) RNA and synthesize substantial amounts of pro-CCK, but differ in their ability to process the prohormone. Seven cultured human neural tumor lines, including two primitive neuroepitheliomas and five neuroblastomas, were screened for CCK gene expression using a CCK C-terminal RIA system, a RIA specific for the carboxy-terminal extension of prepro-CCK, and RNAse protection assays specific for CCK mRNA. RNA derived from the two primitive neuroepitheliomas yielded strong hybridization signals with CCK-specific probes. The five other neuronal tumors were negative for CCK mRNA. The two primitive neuroepitheliomas also synthesized substantial quantities of material reactive in the CCK carboxy-terminal extension RIA system. One of the tumors, Sk-N-Mc, postranslationally processed the CCK prohormonal material poorly, yielding only high mol wt CCK precursors and no immunoreactive CCK. The other, SK-PN-Dw, was able to process the prohormone, producing immunoreactive CCK-like material plus a peptide that immunochemically and chromatographically resembled the intact CCK C-terminal extension peptide. These cultured tumor lines should prove useful in furthering our understanding of the regulation of the CCK gene in human neuronal cells and will also provide an in vitro system for investigation of the posttranslational processing of the CCK preprohormone. In addition, the data demonstrate that screening procedures for examining peptide hormone expression by tumors are most comprehensive when specific molecular genetic probes are employed in addition to standard peptide assay methodology. Finally, these data suggest that CCK production may be a feature of some primitive neuroepitheliomas.

Leukemia after a small dose of radioiodine for metastatic thyroid cancer.

Radioiodine is widely used in the treatment of thyroid cancer. It is one of the most benign forms of therapy for malignancy. Leukemia is a rare complication of 131I therapy, usually occurring after cumulative dosages of more than 800 mCi and with intervals between doses of less than 12 months. We report the occurrence of acute myelogenous leukemia in a 28-yr-old woman 14 months after receiving a total dose of 300 mCi 131I for metastatic follicular thyroid cancer. We also review the published literature of the incidence of leukemia after low dose 131I.

Growth hormone response to growth hormone-releasing hormone in schizophrenic patients.

Ten schizophrenic patients and five normal control subjects were challenged with growth hormone-releasing hormone in a pilot study investigating growth hormone secretion from the pituitary. The results suggest suprapituitary dysfunction in schizophrenia, but replication in a larger study is needed.

Development of Nelson's syndrome in a patient with recurrent Cushing's disease. Analysis of secretory behavior of the pituitary tumor.

A markedly cushingoid 32-year-old man presented to Queens Hospital Center with headache, hyperpigmentation, and visual field loss. Twelve years earlier, he had undergone subtotal adrenalectomy for Cushing's disease, but symptoms of hypercortisolemia promptly recurred. Workup revealed the presence of a large, expanding intrasellar mass, plasma ACTH levels between 3,000 and 10,000 pg/ml, and markedly elevated cortisol levels. The secretion of ACTH (mainly ACTH 1-39-like peptide) by the pituitary tumor showed neither diurnal periodicity nor response to a variety of pharmacologic agents known to affect ACTH secretion. The patient demonstrates a rarely observed presentation of Nelson's syndrome, with aggressive adrenotropic pituitary tumor growth even in the presence of chronic hypercortisolemia.

Synthetic guinea pig insulin B-chain C-terminal decapeptide: a novel immunogen for generating immunocytochemical-grade antisera.

Antisera to guinea pig insulin are not commonly available, largely because of the short supply and limited immunogenicity of the intact hormone. To overcome these problems we have employed a novel reagent, synthetic guinea pig insulin B-chain C-terminal decapeptide, as a hapten for raising antibodies that react with intact guinea pig insulin. The decapeptide, coupled to bovine serum albumin, was successfully used as an immunogen in rabbits. The resulting anti-serum was employed for immunocytochemical staining of guinea pig insulin in pancreatic sections. The specificity of the staining was verified by both pre-absorption and pre-immune serum controls. The utility of this new antiserum for investigations of guinea pig insulin physiology is discussed.

Retinal degeneration 6 (rd6): a new mouse model for human retinitis punctata albescens.

PURPOSE: To characterize the genetics and phenotype of a new mouse mutant with retinal degeneration, rd6, that is associated with extensive, scattered, small white retinal dots seen ophthalmoscopically. METHODS: The phenotype was characterized using ophthalmoscopy, fundus photography, electroretinography, light microscopy, immunocytochemistry, and electron microscopy. Genetic characterization and linkage analysis studies were performed using standard methods. RESULTS: The inheritance pattern of rd6 is autosomal recessive. Linkage analysis mapped rd6 to mouse Chromosome 9 approximately 24 cM from the centromere, suggesting that the human homolog may be on chromosome 11q23. Ophthalmoscopic examination of mice homozygous for rd6 revealed discrete subretinal spots oriented in a regular pattern across the retina. The retinal spots appeared by 8 to 10 weeks of age and persisted through advanced stages of retinal degeneration. Histologic examination revealed large cells in the subretinal space, typically juxtaposed to the retinal pigment epithelium. The white dots seen on fundus examination corresponded both in distribution and size to these large cells. By 3 months of age, the cells were filled with membranous profiles, lipofuscin-like material, and pigment. These cells reacted strongly with an antibody directed against a mouse macrophage-associated antigen. Photoreceptor cells progressively degenerated with age, and an abnormal electroretinogram was initially detected between 1 and 2 months of age. CONCLUSIONS: The fundi of mice homozygous for rd6 exhibit phenotypic similarities to the human flecked retinal disorder retinitis punctata albescens. Thus, rd6/rd6 mice may be a model for understanding the etiology of this or similar disorders. The relationship between the aberrant subretinal cells and the concomitant photoreceptor degeneration remains to be established.

The spatial distribution of pancreatic islets follows a universal power law.

In a previous report we quantified the apparently complex and irregular distribution of pancreatic islets in the guinea pig by showing that they formed a set of cluster or correlation dimension approximately 2.5. Here we show that this distribution holds in a wide range of mammalian species and through ontogenetic development in the guinea pig. These results strongly suggest that islet formation follows an iterative or fractal rule which is universal among mammals.

Statistical geometry of pancreatic islets.

Quantitative histomorphometric studies of the dynamics of growth and development of pancreatic islets in normal and pathological states pose substantial methodological and conceptual problems. We address these problems with the geometry of random fractals, and apply our methods to the analysis of islet regeneration in the alloxan-treated guinea-pig. In both experimental islet-regenerated and control animals, islet centres are found to cluster in similar fractal subsets of dimension strictly less than 3, in agreement with the postulated origin of islets along a system of ductules, and suggesting that regeneration follows the same mathematical dynamics as original islet formation.

Lack of association of higher insulin levels with diffuse atherosclerotic coronary artery disease in nondiabetics.

Since there is experimental evidence that insulin promotes atherosclerosis, we tested the hypothesis that insulin levels are higher in patients with diffuse atherosclerotic coronary artery disease by measuring insulin levels in 46 nondiabetic patients with angiographically defined diffuse coronary artery disease and 46 normal controls with angiographically normal coronary arteries. Fasting insulin levels were similar in both groups of patients: 7.70 +/- 5.77 microU/mL in those with diffuse coronary disease versus 7.39 +/- 5.01 microU/mL in controls. Also, insulin levels drawn 1 and 2 h after oral glucose challenge were not significantly different in patients with diffuse disease (48.78 +/- 32.46 microU/mL and 42.26 +/- 32.38 microU/mL, respectively) compared with patients with normal coronary arteries (51.03 +/- 28.01 microU/mL and 43.79 +/- 31.62 microU/mL, respectively). We conclude that insulin probably does not promote clinical atherosclerosis in nondiabetics.

Studies of antiproteolytic effects of glyburide on rat L6 myoblasts: comparisons with insulin.

Cultured L6 myoblasts afford considerable advantages for identifying and studying the insulin-like actions of test substances in a muscle-derived line. We have used this system to examine the interaction of the oral hypoglycemic sulfonylurea glyburide with bovine insulin on protein degradation and synthesis as well as on thymidine incorporation (as a measure of DNA synthesis) in these cells. Bovine insulin, at doses of 0.1 microgram/mL to 10 micrograms/mL, produced a dose-dependent inhibition of protein degradation (measured by release of trichloracetic acid (TCA)-soluble 14C-tyrosine from myoblasts into the culture medium) and increase in total protein content in the cultured myoblasts. At concentrations of 10 micrograms/mL, insulin achieved its maximal suppression of protein degradation (by nearly 50%) and increased cellular protein content (by 15%) over levels observed in the absence of added insulin. Glyburide, at concentrations at or above 1 microgram/mL, significantly suppressed protein degradation (up to 14%) and slightly augmented protein content of the cells. The effects of glyburide on protein degradation were additive with those of submaximally but not maximally effective concentrations of insulin, suggesting a common mechanism of action of the compounds. Both insulin and glyburide, at maximally effective doses, significantly depressed protein degradation as early as 2 to 6 hours after exposure. In addition, in a 24-hour labeling experiment, insulin stimulated tyrosine incorporation into TCA-insoluble protein and thymidine incorporation into DNA in the cells, whereas glyburide did not enhance these processes and, under certain conditions, inhibited them. These results demonstrate that glyburide, either alone or in concert with insulin, is capable of significantly inhibiting protein turnover in skeletal muscle-derived cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of a sentinel host system to study the questing behavior of Ixodes spinipalpis and its role in the transmission of Borrelia bissettii, human granulocytic ehrlichiosis, and Babesia microti.

Ixodes spinipalpis maintains Borrelia bissettii spirochetes in Colorado in a cycle involving wood rats and deer mice. This tick has been described as nidicolous, remaining either attached to its rodent hosts or in the rodent nest. Nidicolous ticks pose little risk of pathogen transmission to humans if they do not actively quest for hosts. To investigate the questing potential of I. spinipalpis, sentinel mice were placed in an area where I. spinipalpis had been commonly found on wood rats and deer mice. Concurrently, wild rodent populations were trapped and analyzed for Lyme disease spirochetes, the agent of human granulocytic ehrlichiosis (aoHGE), and Babesia microti. A total of 122 I. spinipalpis larvae and 10 nymphs were found on 19% of 244 sentinel mice. In addition, 4 sentinel mice became infested with Malaraeus telchinus or Orchopeas neotomae fleas. Questing I. spinipalpis were positively associated with woody shrubs and negatively associated with sunny and grassy areas. Four sentinel mice became infected with aoHGE after having been fed upon only by I. spinipalpis larvae. One sentinel mouse became infected with B. bissettii after having an I. spinipalpis nymph feed on it, and one sentinel mouse became coinfected with aoHGE and B. bissettii after it was fed upon by a single I. spinipalpis nymph. These sentinel mouse conversions suggest the possibility that the aoHGE is transovarially transmitted by I. spinipalpis, and that I. spinipalpis is capable of simultaneously transmitting B. bissettii and the aoHGE. The findings that I. spinipalpis quest away from rodent nests and will attach to and infect sentinel mice may be of public health importance. It suggests the potential transmission of the agents of human granulocytic ehrlichiosis and Lyme disease to other hosts by I. spinipalpis, in regions of the western United States where Ixodes pacificus is not found.

The regional and subcellular development of cholecystokinin immunoreactivity in vertebrate brain.

We have previously demonstrated that the development of CCK in rat brain occurs during the first postnatal month. In order to determine whether the appearance of CCK is associated with specific aspects of brain histogenesis, we examined the development of brain CCK immunoreactivity in both precocial and altricial mammals and birds. The two precocial species (guinea pig and chicken) were found to achieve adult CCK concentrations prenatally, while the altricial species (zebra finch and rat) manifested adult brain CCK concentrations only after several weeks of postnatal development. In adulthood, both mammals showed relatively high forebrain CCK concentrations, while the two species of birds manifested much lower forebrain levels. Brainstem levels of CCK were similar in all species studied. In each species, the development of CCK followed a common time course across all major brain areas, although adult brainstem levels of CCK were generally attained shortly before adult forebrain levels. Correlation of our comparative ontogenetic data with known patterns of brain histogenesis indicated that CCK development follows regional neuroblastic proliferation, migration and differentiation, and occurs during or soon after local synaptogenesis. In the rapidly developing precocial chicken brain, CCK production precedes the postnatal gliogenic and myelinogenic increases in brain weight, suggesting that neurogenic production of CCK occurs independently of these non-neuronal maturation events. Subcellular fractionation of developing chicken brain revealed that a substantial fraction of brain CCK is localized in synaptosomes relatively early in embryogenesis; this synaptosomal localization becomes even more pronounced with further brain maturation. This early appearance of CCK in synaptic terminals indicates a correspondingly precocial maturation for the intraneuronal mechanisms subserving peptide cleavage, axonal transport and vesicular insertion, and suggests that CCK may be available for neurotransmission quite early in development. In an analysis of the molecular forms of CCK, gel filtration disclosed no differences between species or different brain areas in the form of CCK present. CCK-8 always predominated in brain, with smaller void volume (pro-CCK) peaks, and negligible amounts of CCK-33. Finally, duodenal CCK (largely CCK-33) appeared much earlier than brain CCK in all species examined, suggesting that the gut and brain CCK systems develop independently of one another.

An analysis of spirochete load, strain, and pathology in a model of tick-transmitted Lyme borreliosis.

Four laboratory-grown, low-passage isolates of Borrelia burgdorferi sensu stricto, B31, JD-1, 910255, and N40, were incorporated into Ixodes scapularis ticks to examine the pathogenesis of these isolates in mice after tick transmission. All isolates induced multifocal, lymphoid nodular cystitis, subacute, multifocal, necrotizing myocarditis, and a localized periostitis and arthritis of the femorotibial joint 6-18 weeks after tick infestation. In terms of the number of mice that demonstrated pathology in bladder, heart, and joint, the highest incidence of lesions occurred 12 weeks after tick bite. Utilizing the Taqman quantitative polymerase chain reaction (q-PCR) fluorogenic detection technology to amplify a conserved region of the flagellin gene, a trend was demonstrated between the number of spirochetes in tissue with duration of pathology. The q-PCR assay developed for this study was sensitive and could reliably measure as few as 1 to 10 spirochetes in the target tissues tested. A higher percentage of B31- and N40-infected mice (92 and 100%, respectively) developed myocarditis than JD-1- or 910255-infected mice (67 and 46%, respectively) 12 weeks after tick bite. The amount of spirochetal DNA that could be amplified for heart at this time point was not statistically different between isolates, indicating a difference in virulence between B31 and N40 relative to JD-1 and 910225. N40-infected mice demonstrated a significantly higher spirochete load (an average of 1.23 spirochetes/mg of tissue, p = 0.045) in femorotibial joints 18 weeks after infection, with 60% of these mice maintaining lesions compared with those infected with B31 (13%), JD-1 (25%), or 910255 (50%), which averaged <0.5 spirochetes/mg of tissue. This mouse model of Lyme borreliosis, including the ability to monitor lesion development and spirochete load, can facilitate the testing of therapeutic regimens for the later stages of tick-transmitted Lyme disease and help investigate aspects of the immunopathogenesis of lesion development.