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Detection of DNA sequences in Plasmodium berghei by means of in situ hybridization.

A non-radioactive in situ hybridization technique, used to map unique DNA sequences to plant chromosomes, has been adapted for the localization of specific DNA sequences in nuclei of Plasmodium berghei. After hybridization using probes labeled with biotin-11-dUTP, the formed DNA/DNA hybrids were detected by fluorescence microscopy using a specific double-layer antibody technique. Besides its high resolution, this procedure is characterized by a high sensitivity, allowing the detection of a unique sequence as small as 2.5 kb.

Animals↗

Effects of muscle stimulation in situ in pregnant rats.

Effects of muscle stimulation were studied in the perfused hindleg of late pregnant and age-matched control rats. Muscle stimulation increased tissue glucose uptake in both groups with a tendency towards higher values in the pregnant rats. The stimulation-induced lactate production was markedly larger in pregnant than in control rats. In the pregnant rats muscle tension output decreased more rapidly and vascular resistance was greater than in non-pregnant group. The decline of tension, accompanied by the enhanced lactate production of contracting muscles, may suggest a reduced muscle oxidative potential in late pregnancy and/or delayed adaptation of the local blood flow. The greater vascular resistance found during muscle stimulation in pregnant animals might have been due to the persistent local action of estrogens. The data obtained indicate that pregnancy modifies skeletal muscle function during stimulation in situ.

Animals↗

Handling the emotional response to disaster: the case for American Red Cross/community mental health collaboration.

This paper presents the theory, rationale, and a working model for integrating professional resources at the time of a disaster. Red Cross and community mental health agencies have worked for too long in parallel. A collaborative approach to meeting the emotional needs of disaster victims makes optimal use of indigenous resources; provides skill and knowledge normally lacking in each agency alone; and helps provide, through early intervention, a preventive factor to thwart long term community distress. Options for maintenance of readiness, local or regional adaptation and piloting are also discussed.

Adjustment Disorders↗

Clinical score card for diagnosis of group A streptococcal sore throat.

OBJECTIVE: Group A beta hemolytic streptococcus (GAS) sore throat primarily occurs among children in 5-15 years age group, and if not treated appropriately causes rheumatic fever/rheumatic heart disease (RF/RHD). Present study was aimed at validation of a clinical scoring system for diagnosis of GAS. METHODS: Five hundred and thirty six children in 5-15 years age group were enrolled by systematic random selection of households from a peri-urban slum of Chandigarh. They were visited fortnightly at their home for one year to record signs and symptoms of cough and cold. Throat swabs were collected in 918 episodes, of which 123 (13.4%) were GAS culture positive. RESULT: Significant association of GAS was found with pain in the throat, enlarged tonsils, pharyngeal erythema and tender cervical lymphadenopathy. According to the percentage positivity of GAS culture, weighted scores were assigned to age of the child, season of occurrence, fever, size of tonsil, pharyngeal erythema and exudate, lymphadenopathy and pain in throat. Combinations of various symptoms and signs gave sensitivity of 86-89% and specificity of 83-89% whereas clinical score of 15 or more had 91% sensitivity and 98% specificity for diagnosis of GAS pharyngitis. CONCLUSION: As the level of clinical acumen and prevalence of GAS may differ in different primary care settings of the country, the proposed scoring system should be validated and adapted to suit local conditions before establishing it in the primary prophylaxis strategy to prevention of RF/RHD.

Acute Disease↗

A review of boron effects in the environment.

Boron (B) is a naturally occurring element that is found in the form of borates in the oceans, sedimentary rocks, coal, shale, and in some soils. Borates are released naturally into the atmosphere and aquatic environment from oceans, geothermal steams, and weathering of clay-rich sedimentary rocks. B is also released to a lesser extent from anthropogenic sources. B concentrations in air range from <0.5 to 80 ng/m3 with an average of 20 ng/m3, and in soils from 10 to 300 mg/kg with an average of 30 mg/kg. Concentrations of B in surface freshwaters are typically <0.1-0.5 mg/L; much higher concentrations are measured in a few areas, depending on the geochemical nature of the drainage catchment. B accumulates in both aquatic and terrestrial plants, but it does not appear to be biomagnified through the food chain. No observed effect concentrations (NOECs) for aquatic invertebrates tend to be in the range of 6-10 mg B/L with lower values of 1-2 mg/L for community studies. No effect concentrations for fish in natural waters are around 1 mg/L, although lower values have been recorded in reconstituted water. Comparing no effect concentrations with the general ambient environmental levels indicates that the risk to aquatic ecosystems from B is low. In a few B-rich areas, natural levels will be higher; however, there is some indication that organisms may be adapted to the local conditions. B is an essential micronutrient for higher plants with interspecies differences in the levels required for optimum growth. In general, there is a small concentration range between deficiency and toxicity; however, toxicity owing to excess B is much less common in the environment than B deficiency. Levels of B in aquatic plants growing in areas receiving B-rich runoff from irrigated fields are higher than dietary concentrations, which cause effects on the growth of young birds in the laboratory; however, the bioavailability in the field of such plant-accumulated B is uncertain.

Animals↗

[Rheumatoid arthritis - pathogenetic role of neuroendocrine axes and the peripheral nervous system].

BACKGROUND: Local innate and adaptive immune processes are of importance during the acute phase of rheumatoid arthritis (RA). In the advanced inflammatory phase alterations of systemic anti-inflammatory feedback mechanisms might be important features which may support chronic inflammation. ALTERATIONS: Similarly, like in other chronic inflammatory diseases, inadequately low cortisol and androgen serum levels can be detected in RA patients. In addition, there is a marked reduction of anti-inflammatory sympathetic nerve fibers in the inflamed joints paralleled by an enhanced number of pro-inflammatory sensory nerve fibers. Thus, an uncoupling of synergistically acting endocrine and neuronal, anti-inflammatory mechanisms (cortisol, dehydroepiandrosterone, androgens, sympathetic neurotransmitters) and a preponderance of pro-inflammatory mechanisms (estrogens, sensory neurotransmitters) may lead to chronic inflammatory disease. CONCLUSION: From this pathogenetic point of view new therapeutic strategies could be developed for the treatment of patients with RA.

Androgens↗

[Importance of rectal extirpation for the therapy concept of low rectal cancers].

INTRODUCTION: The main objective of surgery of rectal carcinomas is to avoid a permanent colostomy by sphincter-sparing surgical procedures. A variety of different abdominoperineal resection rates is described in the literature. MATERIAL/METHOD: The study was performed in 2000 within the framework of a multicentric study including 282 hospitals. The purpose of the study was to document the quality of diagnosis and therapy for colorectal carcinomas.A total of 9477 patients were included in this study: 3402 suffering from a rectal carcinoma and 6075 suffering from a colon carcinoma. RESULTS: A total of 866 abdominoperineal resections was performed. This corresponds to an abdominoperineal resection rate of 27.4%. In 30.4% of all men and in 23.0% of all women an abdominoperineal resection was performed. Of all tumor patients who underwent abdominoperineal resection, 8.3% had a pT4 carcinoma and 57.5% a pT3 carcinoma. Adapted to the localization of the tumor in the rectum, i.e., the distance of the aboral tumor margin to the anal verge, the following abdominoperineal resection rates were found: <4 cm from the anal verge 84.6%, 4-7.9 cm 43.9%, 8-11.9 cm 5.8%, and 12-16 cm 0.5%.Intraoperative complications occurred in 11.8%, specific postoperative complications in 33.1%, and general postoperative complications in 27.4% of the patients. The postoperative lethality was 2.8%. The mean postoperative hospital stay was 21.7 days. Logistic regression identified the body mass index, gender, the distance of the carcinoma from the anal verge, and the T category as independent factors influencing the abdominoperineal resection rate. DISCUSSION: Despite an overall decrease in use, abdominoperineal resection will continue to play an important role for the surgical treatment of low rectal cancers in routine clinical practice in Germany. It will remain an individual decision for each patient whether the tumor and the patient allow sphincter preservation or whether abdominoperineal resection seems to be necessary. According to the results of the present study,a general definition of an abdominoperineal resection rate in an unselected group of patients should be viewed critically.

Adult↗

Retinal regionalization and heterogeneity of butterfly eyes.

The regional characteristics of the eyes of butterflies from different families have been surveyed using epi-illumination microscopy, utilizing the eyeshine visible due to the tapetum situated proximally to the rhabdom. All butterflies studied have a high spatial acuity in the frontal region. The facet diameter varies slightly across the eye, and the interommatidial angle and the eye parameter p are especially large dorsally. Whereas the ommatidial lattice is generally highly regular, the eyeshine colours distinctly depend on the species. Sometimes the eyeshine is locally uniform, but often it is heterogeneous. It is hypothesized that the regional characteristics as well as the local heterogeneity are adaptations that optimize spectral discrimination.

Animals↗

Effect of heat and pH stress in the growth of chickpea mesorhizobia.

The development of rhizobial inoculants requires the selection of isolates that are symbiotically efficient as well as adapted to the local environmental conditions. Our aim was to find indigenous chickpea rhizobia tolerant to adverse environmental conditions, such as temperature and pH. Thirteen isolates of chickpea mesorhizobia from southern Portugal were examined. Tolerance to stress temperatures and pH was evaluated by quantification of bacterial growth at 20-37 degrees C and pH 5-9, respectively. Tolerance to heat shocks was studied by submitting isolates to 46 degrees C and 60 degrees C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein analysis revealed qualitative and quantitative differences when isolates were submitted to temperature stress. A 60-kDa protein was overproduced by all isolates under heat stress. Almost all isolates revealed to be more tolerant to 20 degrees C than to 37 degrees C. A positive correlation was found between the maximum growth pH and the isolate origin soil pH. Generally, isolates more tolerant to temperature stress showed a lower symbiotic efficiency.

Bacterial Proteins↗

The relationship between the insulin-like growth factor-1 system and the oestrogen metabolising enzymes in breast cancer tissue and its adjacent non-cancerous tissue.

AIMS: Previous studies have shown that oestrogen and Insulin-like Growth Factor-1 (IGF-1) act synergistically and cross-stimulatory while the oestrogen receptor (ER) and IGF-1R downstream signalling pathways interact at many levels. We investigate the relationship between the ER, and IGF-1R and their ligands in a series of human breast cancer tissue and adjacent non-cancerous tissue (ANCT). METHODS: A series of 139 pairs of breast cancer tissue and ANCT were obtained and divided into ER positive and ER negative groups based on tumour ER alpha immunostaining. All samples were processed for real-time quantitative-PCR to measure IGF-1, IGF-1R, ER alpha, STS and Cyp-19 mRNA levels. In addition, ER positive MCF-7 and ER negative MDA-MB-231 cell lines were treated separately with IGF-1 and an IGF-1R inhibitor called Tyrphostin AG1024 to see the effects of stimulating and inhibiting the IGF-1R. MCF-7 cell line was also treated with 4-hydroxytamoxifen. The mRNA levels of IGF-1, IGF-1R, ER alpha, STS and Cyp-19 of treated cell lines were measured and compared to those of non-treated controls. Data generated was normalised to Cytokeratin-19 mRNA levels. RESULTS: IGF-1R expression was higher in tumour tissue compared to ANCT (P = 0.038) while IGF-1 expression was marginally higher in ANCT compared to tumour tissue only in the ER positive samples (P = 0.098). ER positive tumours had a higher expression of IGF-1 compared to ER negative tumours (P = 0.001) while IGF-1R, STS and Cyp-19 expression were higher in ER negative tumours (P = 0.000, 0.000 and 0.006 respectively). There was no difference in STS or Cyp-19 expression in tumours or ANCT. Using Spearman's Correlation test, IGF-1 positively correlated with STS, Cyp-19 and ER alpha in ER positive and negative groups (Coefficient = +0.497, +0.662 and +0.651 respectively, P = 0.000 in all). IGF-1R correlated with IGF-1, STS, Cyp-19 and ER alpha only in the ER negative tumours (Coefficient = +0.620, +0.394, +0.692 and +0.662 respectively, P = 0.000, 0.012, 0.000 and 0.000 respectively). In cell lines, IGF-1 treatment led to an increase in the mean expression of IGF-1, IGF-1R, STS and Cyp-19 in both cell lines while ER alpha expression increased only in MCF-7. IGF-1R inhibition caused a decrease in expression of all five genes in MDA-MB-231 but not in the MCF-7 cell line. Treatment with 4-hydroxytamoxifen caused a decrease in expression of all five genes. CONCLUSIONS: IGF-1R is over-expressed in malignant tissue. IGF-1 is expressed at higher levels in ER positive tumours probably as a result of oestrogen stimulation while IGF-1R expression is higher in ER negative samples as an adaptation to lower local IGF-1 levels. An IGF-1 paracrine relationship may exist between tumour and ANCT but for STS and Cyp-19, there may be an autocrine-paracrine relationship. The IGF-1 ligand-receptor system is an important regulator of oestrogen production while oestrogen may be involved in stimulating IGF-1 expression. The expression of oestrogen synthesising enzymes is higher in ER negative breast cancers which may be due to the lack of oestrogen negative feedback or contribution from the overexpression of IGF-1R.

Adult↗

Do young heroin users in Madrid, Barcelona and Seville have sufficient knowledge of the risk factors for unintentional opioid overdose?

To identify the self-perceived reasons for unintentional opioid overdose of young heroin users in three Spanish cities and their agreement with objective risk factors for overdose. Computer-Assisted Personal Interviews (CAPI) were held with 991 street-recruited current heroin users aged 18-30. The general reasons for overdose and the reasons for the last overdose suffered were explored with open-ended (OEQs) and pre-coded questions (PCQs). Limited knowledge of overdose risk factors was defined as mention of fewer than two objective risk factors for unintentional overdose in the OEQ. Univariate, bivariate, and logistic regression methods were used. 77.8% (Seville), 64.9% (Madrid) and 57.2% (Barcelona) of participants have limited knowledge of overdose risk factors. Residence in Seville and not having attended courses or meetings on overdoses were significantly associated with limited knowledge, after adjusting for other factors. The most frequently identified general reasons in OEQ or PCQ were using heroin in large amounts (66.8%), together with tranquilizers (62.0%), adulterated (60.7%), or purer than usual (57.6%). Most reasons were selected more frequently in PCQ than in OEQ, especially rapid injection of the entire dose and using heroin shortly after using tranquilizers or alcohol, by injection, or after a period of abstinence. The results were similar for overdoses suffered by participants. Most young heroin users do not have sufficient knowledge of overdose risk factors, especially the use of heroin by injection, after a period of abstinence, or together with alcohol or methadone. Specific informational or educational programs adapted to the local context are critically needed.

Adolescent↗

Reversible, highly localized alterations in fatty acid metabolism in the chronically ischemic canine myocardium.

Acute myocardial ischemia is accompanied by a marked decrease in the oxidation of fatty acids. Whether similar changes occur during chronic ischemia was studied in 13 beagles. In 10 dogs, an ameroid constrictor was implanted about the left circumflex artery; 3 others served as sham-operated controls. Mitochondrial and peroxisomal fatty acid oxidation of (1-14C)oleate were measured and compared in affected (posterobasal left ventricular endocardial [posterior LV endo]) and unaffected (anteroapical LV epicardial [anterior LV epi]) tissues of control and experimental dogs. Five experimental and 3 control dogs were killed after 3 weeks. The posterior LV endo sections of experimental dogs at 3 weeks showed increased fatty acid oxidation due to peroxisomal (KCN-insensitive) beta oxidation (p less than 0.01). The anterior LV epi sections showed no difference in fatty acid oxidation between sham-operated and experimental dogs. Five dogs were studied after 3 months; fatty acid oxidation of the posterior LV endo section was normal. Thus, under a slowly evolving state of myocardial ischemia, highly localized and reversible adaptive changes in fatty acid oxidation occur that enable the affected tissue to cope with a fatty acid load. When collateralization is accomplished, fatty acid metabolism reverts to normal.

Acyl Coenzyme A↗

Decreased rat brain cytochrome oxidase activity after prolonged hypoxia.

Three weeks of hypoxic exposure results in a spectrum of systemic physiological and local brain tissue adaptations. Cytochrome oxidase histochemistry was used as an indicator of changes in energy demand in response to hypoxia. We found overall cytochrome oxidase activity decreased in hypoxic adapted rats as compared to normoxic control rats. Some regions, notably layer 3 of the frontal cerebral cortex, layer 4 of the parietal sensory barrel fields, lacunosum moleculare hippocampi, and specific nuclei of the rostral ventral medulla, though exhibiting cytochrome oxidase activity decreases of 16-27%, still retain their relatively higher levels of activity. We conclude that there is a hypometabolic component of the rat brain adaptation to continued hypoxia.

Animals↗

Co-expression of receptor mRNA and protein: striatal dopamine and excitatory amino acid subtypes.

Dopamine (DA) is known to modulate the post-synaptic response of the excitatory amino acid (EAA) neurotransmitters in the striatum. Thus the intrinsic neurons in this nucleus are potential sites of cross-interaction between these two systems. The recent isolation of 5 different DA receptor subtypes and more than 20 EAA subunits argues for a complicated functional role for the protein products encoded by these transcripts. The simultaneous detection of cellular mRNA distributions and translated protein products was an initial step to determine differences in post-translational expression at the cellular level of resolution for two of these receptors. The cloned D2 DA receptor subtype and the ionotropic GluR1 EAA receptor subunit were examined by fluorescence in situ transcription (FIST) following hybridization of specific cDNA primers, complementary to the mRNA transcripts encoding these receptors. Nascent extension of the annealed primer using reverse transcriptase was detected after incorporation of fluorescently labeled dUTP. Protein products were visualized by standard immunofluorescence after incubation with anti-peptide antisera that were selective for each receptor protein. The experimental data corroborate previous work describing the regional expression of ligand binding and in situ hybridization detected with radiolabeled probes for the DA and EAA receptor systems in the striatum. The dual fluorescence method can be completed within 2 days and may be adapted to cellular localization of many novel mRNA/protein combinations to examine post-translational processing within thin tissue slices.

Animals↗

Equity in authorship: a strategy for assigning credit when publishing.

Existing guidelines for authorship are lacking in specific detail, differ from discipline to discipline, implicitly or explicitly contradict each other on many issues, and are therefore open to varying interpretation by different people. In order to encourage and facilitate consistency and fair acknowledgement of contributions to publications, a Points System was developed for evaluating 13 categories of contribution by individuals. The system is intended for use in determining who should be listed as authors, and in what sequence names should appear by enabling points "earned" through contribution to be then "spent" in purchasing authorship. The system is based on ratings of perceived values of the contribution types obtained from the staff of a multi-disciplinary research centre, but it can readily be adapted to the local needs and values of any research group. The system is likely to be particularly useful in relation to complex research projects and programs, to which a number of individuals from different disciplines make contributions over time, and/or from which several publications are derived.

Authorship↗

Changes in the distribution of the neuron-specific B-50, neurofilament protein and glial fibrillary acidic proteins following an unilateral mesencephalic lesion in the rat.

Following a unilateral electrolytic lesion in the ventral rat mesencephalon, changes in the immunocytochemical distribution of the neuron-specific B-50, neurofilament (NF) protein and glial fibrillary acidic (GFAP) proteins were studied around the lesion after 0, 3, 10 and 28 days. At all recovery times, the controls displayed on immunostaining with anti-B-50 and anti-neurofilament antibodies, a characteristic pattern of synaptic and neuritic localization of these antigens, whereas anti-GFAP staining revealed a distribution typical for astrocytes. The lesion was characterized by a center of coagulated material that exhibited immunoreactivity to B-50 (BIR) and NF (NFIR), but never GFAP-immunoreactivity. From 3 days on, the center became surrounded by disintegrating cells which were unreactive to the antibodies. The antigen distribution changed temporally, predominantly at the lesion rim. By 10 and 28 days postlesion, additional BIR was observed as punctuate dots in fibers and membranes of neurons. Enhanced NFIR was detected in fibers and cell bodies. Many astrocytes were detected around the lesion rim, forming by 28 days postsurgery a barrier between the lesion cavity and the uninjured tissue. Our study shows that distribution changes in B-50, NF and GFAP around the lesion may indicate local degenerative and adaptative processes as a temporal response to brain trauma.

Animals↗

ROC assessment of compressed wrist radiographs.

The aim of this paper is to validate a compression scheme applied on a medical image database of digitized wrist radiographs. The compression scheme adapts itself to local statistical properties of the images. The diagnostic quality of the reconstructed images is evaluated using a ROC protocol involving five medical experts. The results of this evaluation enable us to validate the compression scheme on this database with a compression ratio of 40 (0.2 bits per pixel).

Humans↗