Compartment analysis of T cell receptor gamma--and immunoglobulin V(H) rearrangements by microdissection in patients with malt lymphoma and chronic gastritis with lymphatic hyperplasia.

The interpretation of clonality within H. pylori-associated gastritis and low-grade MALT lymphoma remains controversial. Due to the observation of MALT lymphoma regression after H. pylori eradication, new definitions concerning the border between benign reactive lesions and malignant gastric lymphoma are needed. Gene rearrangements for immunoglobulin heavy-chain in low-grade MALT lymphoma (N= 12) and H. pylori-associated chronic gastritis with lymphatic hyperplasia (N= 13) were analyzed by microdissection and polymerase chain reaction. Furthermore, T cell receptor-gamma chain rearrangements were analyzed by gene scan analysis. In 11 of 12 cases with initial low-grade MALT lymphoma, intraepithelial and subepithelial B cell rearrangements showed a restricted usage of the immunoglobulin heavy-chain 3. In H. pylori-associated chronic gastritis, the intraepithelial B cell compartment showed an oligoclonal the immunoglobulin heavy-chain rearrangement pattern with a predominance of VH3. The subepithelial compartment did not show any restrictive immunoglobulin heavy-chain gene usage. Additionally a mono- to oligoclonal rearrangement pattern of the T cell receptor-y chain was observed in low-grade MALT lymphoma, whereas an oligoclonal pattem was observed in chronic gastritis. Our data provide evidence that low-grade MALT lymphoma may start within the epithelium and subsequently infiltrate the subepithelial compartment. The observation of a mono- to oligoclonal TCR-gamma rearrangement suggests that an antigen selecting process also takes place within reactive T cells. Combining TCR-gamma gene scan analysis with IgH chain rearrangement analysis might help in discriminating between chronic gastritis and initial MALT lymphoma in questionable cases.     

Prevalence of Depression,Suicidal Ideation,Alcohol Intake and Nicotine Consumption in Rural Central India. The Central India Eye and Medical Study

Background

To investigate the prevalence of depression, suicidal ideations, alcohol and nicotine consumption in adults in an agrarian society mostly unchanged by the effects of urbanization.

Methods

The Central India Eye and Medical Study is a population-based study in rural Central India close to the tribal belt and included 4711 subjects (aged 30+ years). Depression was assessed by the Center for Epidemiologic Studies Depression Scale (CESD), suicidal ideation by six standardized questions, nicotine use by the Fagerstroem Nicotine Tolerance Questionnaire (FTNQ), and alcohol consumption by the Alcohol Use Disorders Identification Test (AUDIT).

Results

Mild to moderate depression (CESD sum score: 15–21) was detected in 1862 (39.6%) individuals (33.5% of men, 44.8 of women), and major depression (CESD sum score >21) in 613 (13.0%) individuals (8.1 of men, 17.3% of women). Suicide attempt was reported by 199 (4.2%) participants and suicidal thoughts during the last 6 months by 238 (5.1%) individuals. There were 887 (18.9%) smokers and smokeless tobacco was consumed by 1968 (41.8%) subjects. Alcohol consumption was reported by 1081 (23.0%) participants; 283 (6.0%) subjects had an AUDIT score ≥8 (hazardous drinking), and 108 (4.63%) subjects a score ≥13 (women) or ≥15 (men) (alcohol dependence).

Conclusions

In rural Central India, prevalence of major depression was comparable to figures reported from other developing countries. Prevalence of smoking and hazardous alcohol consumption was higher than as reported from urban regions. Measures should be taken to address the relatively high prevalence of suicide attempts and thoughts on suicide in rural Central India.     

Influence of tidal volume on pulmonary NO release,tissue lipid peroxidation and surfactant phospholipids

Mechanical stress during ventilation may cause or aggravate acute lung injury. This study investigates the influence of low vs. high tidal volume (V(t)) on factors known to play key roles in acute lung injury: nitric oxide release, eNOS and iNOS gene expression, lipid peroxidation (LPO), and surfactant phospholipids (PL). Isolated rabbit lungs were subjected to one of three ventilation patterns for 135 min (V(t)-PEEP): 6 ml/kg-0 cm H(2)O. 12 ml/kg-0 cm H(2)O 6 ml/kg-5 cm H(2)O, 12 ml/kg-0 cm H(2)O, and 6 ml/kg-5 cm H(2)O resulted in comparable peak inspiratory pressure (PIP). This allowed comparing low and high V(t) without dependence on PIP. Ventilatory patterns did not induce changes in pulmonary artery pressure, vascular permeability (K(f,c)), PIP or pulmonary compliance. High V(t) in comparison with both of the low V(t) groups caused an increase in BALF-nitrite (30.6+/-3.0* vs. 21.4+/-2.2 and 16.2+/-3.3 microM), BALF-PL (1110+/-19* vs. 750+/-68 and 634+/-82 microg/ml), and tissue LPO product accumulation (0.62+/-0.051* vs. 0.48+/-0.052 and 0.43+/-0.031 nmol/mg), *P<0.05 each. Perfusate nitrite and BALF-PL composition (assessed by use of 31P-NMR spectroscopy and MALDI-TOF mass spectrometry) did not differ among the groups. High V(t) ventilation reduced eNOS gene expression but did not affect iNOS expression. The increased release of NO and the accumulation of LPO products may represent early lung injury while elevated BALF-PL may reflect distension-induced surfactant secretion.     

The impact of mitochondrial genome analyses on the understanding of deuterostome phylogeny

Deuterostomia, one of the three major lineages of Bilateria, comprises many well-known animals such as vertebrates, sea squirts, sea stars and sea urchins. Whereas monophyly of Deuterostomia and several subtaxa is well supported, the relationships of these to each other and, hence, deuterostome relationships are still uncertain. To address these issues in deuterostome phylogeny we analyzed datasets comprising more than 300 complete deuterostome mitochondrial genomes. Based on sequence information, the results revealed support for several relationships such as a basal position of Xenoturbella within Deuterostomia or for taxa like Craniota or Ambulacraria, but yielded also problems in some taxa, e.g. Tunicata, Pterobranchia and Ophiuroidea, due to long-branch artifacts. However, within tunicates the relationships are well supported. Variation in the genetic code was also informative and, e.g., supported the taxon Ambulacraria including Pterobranchia.     

Land use and host neighbor identity effects on arbuscular mycorrhizal fungal community composition in focal plant rhizosphere

Arbuscular mycorrhizal fungi (AMF) provide a number of ecosystem services as important members of the soil microbial community. Increasing evidence suggests AMF diversity is at least partially controlled by the identities of plants in the host plant neighborhood. However, much of this evidence comes from greenhouse studies or work in invaded systems dominated by single plant species, and has not been tested in species-rich grasslands. We worked in 67 grasslands spread across the three German Biodiversity Exploratories that are managed primarily as pastures and meadows, and collected data on AMF colonization, AMF richness, AMF community composition, plant diversity, and land use around focal Plantago lanceolata plants. We analyzed the data collected within each Exploratory (ALB Schwäbische Alb, HAI Hainich-Dün, SCH Schorfheide-Chorin) separately, and used variance partitioning to quantify the contribution of land use, host plant neighborhood, and spatial arrangement to the effect on AMF community composition. We performed canonical correspondence analysis to quantify the effect of each factor independently by removing the variation explained by the other factors. AMF colonization declined with increasing land use intensity (LUI) along with concurrent increases in non-AMF, suggesting that the ability of AMF to provide protection from pathogens declined under high LUI. In ALB and HAI mowing frequency and percent cover of additional P. lanceolata in the host plant neighborhood were important for AMF community composition. The similar proportional contribution of land use and host neighborhood to AMF community composition in a focal plant rhizosphere suggests that the diversity of this important group of soil microbes is similarly sensitive to changes at large and small scales.     

C-type natriuretic peptide (CNP) signal peptide fragments are present in the human circulation

Background

Signal peptides may be novel biomarkers in cardiovascular diseases.

Methods

We developed a novel immunoassay to the signal peptide of preproCNP (CNPsp) and used this to document circulating venous concentrations of CNPsp in normal healthy volunteers (n = 109), regional plasma CNPsp concentrations in patients undergoing clinically indicated catheterisation (n = 24) and temporal CNPsp concentrations in patients with ST-elevation myocardial infarction (STEMI) <4 h after symptom onset (n = 8). The structure/sequence of circulating CNPsp was confirmed by tandem mass spectrometry (MS/MS).

Results

In normal human plasma, CNPsp was detectable at levels higher than NT-proCNP (74 ± 17 vs. 20 ± 5.5 pmol/L). There was no correlation between NTproCNP and CNPsp, but plasma concentrations of sibling signal peptides – CNPsp and BNPsp – were strongly correlated (r = 0.532, P < 0.001). In patients undergoing catheterisation, there were significant arterio-venous step-ups in CNPsp concentrations across the heart (P < 0.01) and kidney (P < 0.01). Arterial concentrations of CNPsp significantly correlated with heart rate (r = 0.446, P < 0.05). In STEMI patients, plasma concentrations of CNPsp showed a biphasic elevation pattern between 6 and 12 h after symptom onset, with 12 h values significantly elevated (∼3-fold) compared with levels at presentation (P < 0.05). MS/MS verified circulating CNPsp to be preproCNP(14–23) and preproCNP(16–23) peptides.

Conclusions

This is the first report of a circulating preproCNP derived signal peptide. Given the clear cardiac and renal secretion profiles of CNPsp and its response in STEMI patients, further studies on potential biological functions and biomarker applications of CNPsp in cardiovascular disease are warranted.     

Validating Antibodies to the Cannabinoid CB2 Receptor: Antibody Sensitivity Is Not Evidence of Antibody Specificity

Antibody-based methods for the detection and quantification of membrane integral proteins, in particular, the G protein-coupled receptors (GPCRs), have been plagued with issues of primary antibody specificity. In this report, we investigate one of the most commonly utilized commercial antibodies for the cannabinoid CB2 receptor, a GPCR, using immunoblotting in combination with mass spectrometry. In this way, we were able to develop powerful negative and novel positive controls. By doing this, we are able to demonstrate that it is possible for an antibody to be sensitive for a protein of interest—in this case CB2—but still cross-react with other proteins and therefore lack specificity. Specifically, we were able to use western blotting combined with mass spectrometry to unequivocally identify CB2 protein in over-expressing cell lines. This shows that a common practice of validating antibodies with positive controls only is insufficient to ensure antibody reliability. In addition, our work is the first to develop a label-free method of protein detection using mass spectrometry that, with further refinement, could provide unequivocal identification of CB2 receptor protein in native tissues.     

Nutritional and ecological evaluation of dairy farming systems based on concentrate feeding regimes in semi-arid environments of Jordan

The objective of this study was to evaluate the nutritional and ecological aspects of feeding systems practiced under semi-arid environments in Jordan. Nine dairy farms representing the different dairy farming systems were selected for this study. Feed samples (n = 58), fecal samples (n = 108), and milk samples (n = 78) were collected from the farms and analysed for chemical composition. Feed samples were also analysed for metabolisable energy (ME) contents and in vitro organic matter digestibility according to Hohenheim-Feed-Test. Furthermore, fecal nitrogen concentration was determined to estimate in vivo organic matter digestibility. ME and nutrient intakes were calculated based on the farmer’s estimate of dry matter intake and the analysed composition of the feed ingredients. ME and nutrient intakes were compared to recommended standard values for adequate supply of ME, utilizable crude protein, rumen undegradable crude protein (RUCP), phosphorus (P), and calcium (Ca). Technology Impact Policy Impact Calculation model complemented with a partial life cycle assessment model was used to estimate greenhouse gas emissions of milk production at farm gate. The model predicts CH₄, N₂O and CO₂ gases emitted either directly or indirectly. Average daily energy corrected milk yield (ECM) was 19 kg and ranged between 11 and 27 kg. The mean of ME intake of all farms was 184 MJ/d with a range between 115 and 225 MJ/d. Intake of RUCP was lower than the standard requirements in six farms ranging between 19 and 137 g/d, was higher (32 and 93 g/d) in two farms, and matched the requirements in one farm. P intake was higher than the requirements in all farms (mean oversupply = 19 g/d) and ranged between 3 and 30 g/d. Ca intake was significantly below the requirements in small scale farms. Milk nitrogen efficiency N-_eff (milk N/intake N) varied between 19% and 28% and was mainly driven by the level of milk yield. Total CO₂ equivalent (CO₂ equ) emission ranged between 0.90 and 1.88 kg CO₂/kg ECM milk, where the enteric and manure CH₄ contributed to 52% of the total CO₂ equ emissions, followed by the indirect emissions of N₂O and the direct emissions of CO₂ gases which comprises 17% and 15%, respectively, from total CO₂ equ emissions. Emissions per kg of milk were significantly driven by the level of milk production (r² = 0.93) and of eDMI (r² = 0.88), while the total emissions were not influenced by diet composition. A difference of 16 kg ECM/d in milk yield, 9% in N-_eff and of 0.9 kg CO₂ equ/kg in ECM milk observed between low and high yielding animals. To improve the nutritional status of the animals, protein requirements have to be met. Furthermore, low price by-products with a low carbon credit should be included in the diets to replace the high proportion of imported concentrate feeds and consequently improve the economic situation of dairy farms and mitigate CO₂ equ emissions.