The role of nigral projections to the thalamus in drug-induced circling behaviour in the rat

Unilateral injection of 6-hydroxydopamine (6-OHDA) into the ascending nigrostriatal pathway caused contraversive circling to apomorphine and ipsiversive circling to amphetamine respectively. An electrolesion of the ventromedial thalamic nucleus on the same side as the 6-OHDA lesion reduced apomorphine-induced circling, but not that to amphetamine. An electrolesion of the ventromedial thalamic nucleus on the side opposite to the 6-OHDA lesion reduced amphetamine circling but not that to apomorphine. Bilateral electrolesions of the ventromedial thalamic nucleus reduced neither apomorphine- nor amphetamine-induced circling. Electrolytic lesions of the parafascicular thalamic nucleus did not reduce apomorphine- or amphetamine-induced circling in animals with a unilateral 6-OHDA lesion of the nigrostriatal pathway. Knife cuts rostral and dorsal to the substantia nigra did not attenuate circling induced by injection of muscimol into the substantia nigra. Circling due to activition of nigral output pathways can be mediated by descending nigro-reticular pathways.     

Inhibition of normal allogeneic lymphocyte mitogenesis by a factor released from human tumor cells in culture

Summary Human tumor and normal cell lines in culture were examined for the release of factors capable of inhibiting lymphocyte blastogenesis. Supernatants from tumor cell cultures of melanoma, carcinoma (lung, colon, breast), sarcoma, and normal fibroblasts inhibited normal lymphocyte response to PHA. Only supernatants from the tumor cell lines C1 (colon carcinoma), 734B (breast carcinoma), and 231 (breast carcinoma) were found to inhibit both PHA and ConA responses significantly. The two breast carcinoma cell lines, 734B and 231, which also were capable of inhibiting lymphocyte responses to PPD and alloantigens, were investigated further. The inhibition of lymphocyte blastogenesis caused by the supernatant of these two cell lines could not be overcome by the addition of added mitogen. Further experiments showed that the inhibition was not due to nutrient deficiencies and the supernatants were not directly toxic to the lymphocyte cultures as judged by trypan blue exclusion.     

Complete nucleotide and derived amino acid sequence of cDNA encoding the mitochondrial uncoupling protein of rat brown adipose tissue: lack of a mitochondrial targeting presequence.

A cDNA clone spanning the entire amino acid sequence of the nuclear-encoded uncoupling protein of rat brown adipose tissue mitochondria has been isolated and sequenced. With the exception of the N-terminal methionine the deduced N-terminus of the newly synthesized uncoupling protein is identical to the N-terminal 30 amino acids of the native uncoupling protein as determined by protein sequencing. This proves that the protein contains no N-terminal mitochondrial targeting prepiece and that a targeting region must reside within the amino acid sequence of the mature protein.     

Plasma methylglyoxal and glyoxal are elevated and related to early membrane alteration in young, complication-free patients with Type 1 diabetes

The reactive aldehydes methylglyoxal and glyoxal, arise from enzymatic and non-enzymatic degradation of glucose, lipid and protein catabolism, and lipid peroxidation. In Type 1 diabetes mellitus (T1DM) where hyperglycemia, oxidative stress, and lipid peroxidation are common, these aldehydes may be elevated. These aldehydes form advanced glycation end products (AGEs) with proteins that are implicated in diabetic complications. We measured plasma methylglyoxal and glyoxal in young, complication-free T1DM patients and assessed activity of the ubiquitous membrane enzyme, Na⁺/K⁺ ATPase. A total of 56 patients with TIDM (DM group), 6–22 years, and 18 non-diabetics (ND group), 6–21 years, were enrolled. Mean plasma A1C (%) was higher in the DM group (8.5 ± 1.3) as compared to the ND group (5.0 ± 0.3). Using a novel liquid chromatography-mass spectrophotometry method, we found that mean plasma methylglyoxal (nmol/l) and glyoxal levels (nmol/l), respectively, were higher in the DM group (841.7 ± 237.7, 1051.8 ± 515.2) versus the ND group (439.2 ± 90.1, 328.2 ± 207.5). Erythrocyte membrane Na⁺/K⁺ ATPase activity (nmol NADH oxidized/min/mg protein) was elevated in the DM group (4.47 ± 0.98) compared to the ND group (2.16 ± 0.59). A1C correlated with plasma methylglyoxal and glyoxal, and both aldehydes correlated with each other. A high correlation of A1C with Na⁺/K⁺ ATPase activity, and a regression analysis showing A1C as a good predictor of activity of this enzyme, point to a role for glucose in membrane alteration. In complication-free patients, increased plasma methylglyoxal, plasma glyoxal, and erythrocyte Na⁺/K⁺ ATPase activity may foretell future diabetic complications, and emphasize a need for aggressive management.     

Development of Quantitative Real-Time PCR Assays for Detection and Quantification of Surrogate Biological Warfare Agents in Building Debris and Leachate

Evaluation of the fate and transport of biological warfare (BW) agents in landfills requires the development of specific and sensitive detection assays. The objective of the current study was to develop and validate SYBR green quantitative real-time PCR (Q-PCR) assays for the specific detection and quantification of surrogate BW agents in synthetic building debris (SBD) and leachate. Bacillus atrophaeus (vegetative cells and spores) and Serratia marcescens were used as surrogates for Bacillus anthracis (anthrax) and Yersinia pestis (plague), respectively. The targets for SYBR green Q-PCR assays were the 16S-23S rRNA intergenic transcribed spacer (ITS) region and recA gene for B. atrophaeus and the gyrB, wzm, and recA genes for S. marcescens. All assays showed high specificity when tested against 5 ng of closely related Bacillus and Serratia nontarget DNA from 21 organisms. Several spore lysis methods that include a combination of one or more of freeze-thaw cycles, chemical lysis, hot detergent treatment, bead beat homogenization, and sonication were evaluated. All methods tested showed similar threshold cycle values. The limit of detection of the developed Q-PCR assays was determined using DNA extracted from a pure bacterial culture and DNA extracted from sterile water, leachate, and SBD samples spiked with increasing quantities of surrogates. The limit of detection for B. atrophaeus genomic DNA using the ITS and B. atrophaeus recA Q-PCR assays was 7.5 fg per PCR. The limits of detection of S. marcescens genomic DNA using the gyrB, wzm, and S. marcescens recA Q-PCR assays were 7.5 fg, 75 fg, and 7.5 fg per PCR, respectively. Quantification of B. atrophaeus vegetative cells and spores was linear (R² > 0.98) over a 7-log-unit dynamic range down to 10¹ B. atrophaeus cells or spores. Quantification of S. marcescens (R² > 0.98) was linear over a 6-log-unit dynamic range down to 10² S. marcescens cells. The developed Q-PCR assays are highly specific and sensitive and can be used for monitoring the fate and transport of the BW surrogates B. atrophaeus and S. marcescens in building debris and leachate.     

Rhizosphere carboxylate concentrations of chickpea are affected by genotype and soil type

We investigated whether concentrations of carboxylates in the rhizosphere of chickpea (Cicer arietinum L.) roots were related to soil phosphorus levels. In a field experiment, cultivar Sona was grown at two P levels on eight soil types at three locations. There were large differences in extractable (0.2 mM CaCl₂) rhizosphere carboxylate concentrations amongst the locations. The effect of P fertiliser was variable and carboxylate concentrations depended on soil type. To examine the effect of soil P in more detail, a glasshouse experiment was carried out, in which three cultivars (Heera, Sona and Tyson) were grown at four P levels on one soil type. The biomass of chickpea plants increased with increasing P level of the soil, and the root mass ratio decreased at the highest soil P level. However, rhizosphere concentrations of the carboxylates malonate, malate and citrate did not differ significantly between P treatments. This implied that there was no simple relation between available P and root exudation rates, in contrast to earlier results in studies using hydroponics. Cultivars differed in carboxylate concentration pattern: Sona and Tyson showed a tendency towards increased rhizosphere carboxylate concentrations at the second harvest, whereas the carboxylate concentration of Heera tended to decrease. It is hypothesised that chickpea roots always exude a basal level of carboxylates into the rhizosphere. They only increase carboxylate exudation considerably when the P availability is extremely low, which may occur in soils that strongly bind P.     

Gender effects on trapezius surface EMG during delayed onset muscle soreness due to eccentric shoulder exercise.

The purpose of this study was to investigate gender-specific motor control strategies during eccentric exercise and delayed onset muscle soreness (DOMS) in the shoulder region. Twelve healthy males and females participated in the study. Eccentric shoulder exercises were conducted on the dominant shoulder while the other side served as control. The exerted force, range of shoulder elevation, rating of perceived exertion, pain intensity, and surface electromyography (EMG) from the trapezius muscles were recorded and analyzed. A significant decrease in exerted force during exercise was only found in males despite similar rating of perceived exertion among genders. During eccentric exercise: males showed increasing root mean square (RMS) of the EMG while a decrease occurred for females, no difference between genders in mean power frequency of the EMG were seen. During static and dynamic contractions: no differences between genders in pain intensity or RMS were observed; RMS of the exercised side were lower than that of the control side (P<0.05) at 24 h after exercise. The results indicated a more prominent muscle fatigue resistance in females compared with males and mobilization of different muscle activation strategies during eccentric exercise. A protective adaptation to DOMS, i.e. decrease in RMS values was found with no gender differences.     

Determining the Phylogenetic and Phylogeographic Origin of Highly Pathogenic Avian Influenza (H7N3) in Mexico

Highly pathogenic (HP) avian influenza virus (AIV) H7N3 outbreaks occurred 3 times in the Americas in the past 10 years and caused severe economic loss in the affected regions. In June/July 2012, new HP H7N3 outbreaks occurred at commercial farms in Jalisco, Mexico. Outbreaks continued to be identified in neighbouring states in Mexico till August 2013. To explore the origin of this outbreak, time resolved phylogenetic trees were generated from the eight segments of full-length AIV sequences in North America using BEAST. Location, subtype, avian host species and pathogenicity were modelled as discrete traits upon the trees using continuous time Markov chains. A further joint analysis among segments was performed using a hierarchical phylogenetic model (HPM) which allowed trait rates (location, subtype, host species) to be jointly inferred across different segments. The complete spatial diffusion process was visualised through virtual globe software. Our result indicated the Mexico HP H7N3 originated from the large North America low pathogenicity AIV pool through complicated reassortment events. Different segments were contributed by wild waterfowl from different N. American flyways. Five of the eight segments (HA, NA, NP, M, NS) were introduced from wild birds migrating along the central North American flyway, and PB2, PB1 and PA were introduced via the western North American flyway. These results highlight a potential role for Mexico as a hotspot of virus reassortment as it is where wild birds from different migration routes mix during the winter.