The role of the S1 binding site of carboxypeptidase M in substrate specificity and turn-over

The influence of the P1 amino acid on the substrate selectivity, the catalytic parameters K(m) and k(cat), of carboxypeptidase M (CPM) (E.C. 3.4.17.12) was systematically studied using a series of benzoyl-Xaa-Arg substrates. CPM had the highest catalytic efficiency (k(cat)/K(m)) for substrates with Met, Ala and aromatic amino acids in the penultimate position and the lowest with amino acids with branched side-chains. Substrates with Pro in P1 were not cleaved in similar conditions. The P1 substrate preference of CPM differed from that of two other members of the carboxypeptidase family, CPN (CPN/CPE subfamily) and CPB (CPA/CPB subfamily). Aromatic P1 residues discriminated most between CPM and CPN. The type of P2 residue also influenced the k(cat) and K(m) of CPM. Extending the substrate up to P7 had little effect on the catalytic parameters. The substrates were modelled in the active site of CPM. The results indicate that P1-S1 interactions play a role in substrate binding and turn-over.     

Dopamine D1-like receptor-mediated inhibition of Cl/HCO3- exchanger activity in rat intestinal epithelial IEC-6 cells is regulated by G protein-coupled receptor kinase 6 (GRK 6).

The present study investigated the effect of dopamine D1-like receptor stimulation on the Cl-/HCO3- exchange activity in rat intestinal epithelial IEC-6 cells. The Cl-/HCO3- exchange activity was found to be a chloride-dependent, DIDS-sensitive and niflumate-insensitive process. The presence of the SLC26A6 anion exchanger was detected by both RT-PCR and immunoblotting analysis in IEC-6 cells, in which three different small interfering RNAs (siRNAs) targeting SLC26A6 markedly inhibited Cl-/HCO3- exchange. Activation of dopamine D1-like receptors with SKF 38393 inhibited Cl-/HCO3- exchanger activity, this being antagonized by the D1 selective antagonist SKF 83566. However, effects of SKF 38393 were maximal at 5 min of exposure to the agonist and rapidly diminished with no effect at 15 min, suggestive of agonist-induced desensitization of D1-like receptors. Pretreatment of cells with heparin, a non-selective inhibitor of G protein-coupled receptor kinases (GRKs), prevented the observed attenuation of SKF 38393-induced inhibition of Cl-/HCO3- exchange. Overnight pretreatment with anti-GRK6A and anti-GRK6B, but not with anti-GRK4 antibodies, prevented the loss of SKF 38393-mediated effects. Both PKA and PKC signaling pathways participate in SKF 38393-mediated inhibition of Cl-/HCO3- exchange. These findings suggest that SLC26A6 is at least one of the anion exchanger's family members responsible for Cl-/HCO3- exchange in IEC-6 cells. Dopamine D1 receptors in IEC-6 rapidly desensitize to D1-like agonist stimulation and GRK 6, but not GRK 4, appear to be involved in agonist-mediated responsiveness and desensitization.     

Relative contribution of vegetation types to regional biodiversity in Central Zagross forests of Iran

Managers in conservation biology are continually faced with the dilemma of needing to demonstrate which areas should receive conservation priority based on the diversity of species contained. Darenasab (Hashtadpahloo) forest catchment with dominated oak species in Mediterranean forests of Zagross in Iran was chosen as a case study. In order to estimate plant species richness in different vegetation types at landscape level, field method that sample both trees and herbs strata simultaneously (modified multi-scale Whittaker plots) was used to make species–area curves. Twenty-one modified multi-scale Whittaker plots (250 m² area) were located randomly in four vegetation types. Three species–area, species–log (area) and log (species)–log (area) curves models were constructed. The log (species)–log (area) model had the highest adjusted r² among others. Based on Jaccard’s coefficient, the pure oak vegetation type was the most heterogeneous (22% overlap), and the cushion plants vegetation type is the most homogeneous (29% overlap). The slope of species–area curves had the least range (0.05) and the slope of species–log (area) curves had the largest range (4.38). When the slopes of species–log (area) curves divided by mean Jaccard’s coefficient, the species–log (area) curves estimated values closest to those observed. The index of vegetation types contribution to regional diversity for determining conservation priority in off-reserve area, based on ranking of the observed mean species per plots, slopes of the species–log (area) curves, mean of Jaccard’s coefficient, mean unique species per plot, and the number of threatened species in each vegetation types were calculated. This composite index may provide a scientific method to rank vegetation types with high conservation value.     

A new species of Haplothrips from southern Iran (Thysanoptera,Phlaeothripidae)

Haplothrips herajius sp. n. is described from leaves and flowers of a species of Suaeda in the south of Fars Province, Iran. This is the second Iranian species of Haplothrips with the unusual character state of extra setae on the metanotum. Information on variation in color and structure of the new species is provided. The similarities and host plant associations of this new species and Haplothrips kermanensis are discussed,as both are phytophagous on species of Chenopodiaceae.     

Sublethal toxicity of two insect growth regulators on Habrobracon hebetor (Hymenoptera: Braconidae) under laboratory conditions

The sublethal effects of lufenuron and lufenuron?+?fenoxycarb were tested on the adult stage of the parasitoid wasp Habrobracon hebetor under laboratory conditions. Bioassay tests were carried out in the incubator at a temperature of 26?±?1?°C, relative humidity of 65?±?10% and photoperiod of 16:8 (L:D). Exposure cages were used for the experiments. Tests were conducted in three replications. Young female wasps were exposed to the residues of the pesticides’ sublethal concentrations (LC₂₅) after 1, 24, 48 and 72 h, and then their mortalities were recorded. Results showed that both insecticide treatments had significant differences compared to the control, but not between themselves (df?=?1, F?=?29.21, α?<?.0001). It was also revealed that the time between insecticides application and H. hebetor release would significantly affect the survival of searching parasitoids (df?=?3, F?=?237.01, α?<?.0001). In this regard, the releasing programme should be postponed for at least 24 h after using these insect growth regulators.     

Life-history parameters of the western flower thrips,Frankliniella occidentalis,are affected by host plant salinity stress

Salinization, as one of the foremost abiotic stresses, is an intensifying problem in many agroecosystems. Climatic changes, along with altering land use and also salinity of irrigation water all lead to enhanced soil salinity in agricultural lands. Changes in plant characteristics, as a result of raising soil salinity, may impose bottom-up impact on plant-feeding insects. We assessed the bottom-up impact of salinity stress on demographic traits of the western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), on cherry tomato, Solanum lycopersicum L. var. cerasiforme (Solanaceae) plants under greenhouse conditions (27 ± 2 °C, 65 ± 5% r.h., and L16:D8 photoperiod). Our results indicated that salinity stress interfered with the immature development period, adult longevity, and sex ratio of WFT. Salinity stress biased the sex ratio in favor of males. Significant concentration-dependent differences were observed in the intrinsic (r) and finite (λ) increase rates and the net reproduction rate (R₀) of WFT at different salinity levels. Salinity adversely influenced WFT development; nonetheless, population projection forecasted an ascending WFT population growth under moderate salinity stress of 100 mM (2.8 dS m⁻¹ of NaCl), whereas severe salinity stress of 150 mM (4.7 dS m⁻¹ of NaCl) resulted in remarkable fitness costs in WFT. This study demonstrates that WFT has the potential to become problematic in regions with moderate salinity. Therefore, it might exacerbate the detrimental impact of salinity on tomato production. The current survey provides information on the abundance of WFT on saline-stressed tomato plants, thereby contributing to developing environmentally friendly measures to manage this notorious species in ecosystems under salinity stress.     

Spatiotemporal dynamics of re-innervation and hyperinnervation patterns by uninjured CGRP fibers in the rat foot sole epidermis after nerve injury

Background

Minocycline prevents the development of neuropathic and inflammatory pain by inhibiting microglial activation and postsynaptic currents. But, how minocycline obviates acute visceral pain is unclear. The present study investigated whether minocycline had an any antinociceptive effect on acetic acid-induced acute abdominal pain after intraperitoneal (i.p.) administration of saline or minocycline 1 hour before acetic acid injection (1.0%, 250 ??l, i.p.).

Results

Minocycline (4, 10, or 40 mg/kg) significantly decreased acetic acid-induced nociception (0-60 minutes post-injection) and the enhancement in the number of c-Fos positive cells in the T5-L2 spinal cord induced by acetic acid injection. Also, the expression of spinal phosphorylated extracellular signal-regulated kinase (p-ERK) induced by acetic acid was reduced by minocycline pre-administration. Interestingly, intrathecal introduction of PD98059, an ERK upstream kinase inhibitor, markedly blocked the acetic acid-stimulated pain responses.

Conclusions

These results demonstrate that minocycline effectively inhibits acetic acid-induced acute abdominal nociception via the inhibition of neuronal p-ERK expression in the spinal cord, and that minocycline may have therapeutic potential in suppressing acute abdominal pain.     

<Emphasis Type="Italic">Lactobacillus salivarius</Emphasis> NK02: a Potent Probiotic for Clinical Application in Mouthwash

A specific strain of naturally occurring oral lactobacilli was isolated and identified based on morphological, biochemical, and 16S rRNA gene sequencing. The phylogenetic affiliation of the isolate confirmed that the NK02 strain had close association with the Lactobacillus salivarius. An effective mouthwash was developed for treatment of periodontitis and suppression of the indicator bacterium Aggregatibacter actinomycetemcomitans which is an obvious pathogen of periodontal disease. The mouthwash containing L. salivarius NK02 was tested at a dose level of 10⁸ (colony forming units (CFU) ml^?1), monitoring over a period of 4 weeks. The study was a randomized double-blind placebo control trial, and the patients were treated in two groups of control and test by using scaling and root planing (SRP) + placebo and scaling and root planing (SRP) + probiotic, respectively. It appeared that the probiotic mouthwash was able to inhibit the bacterial growth on both saliva and sub-gingival crevice and exhibited antibacterial activity against A. actinomycetemcomitans. The results also showed that SRP+ probiotic treatment led to a significant decrease of gingival index (GI) and bleeding on probing (BOP) compared with that of SRP + placebo for the probiotic group. The rate of decrease in pocket depth was displayed in the group with SRP + probiotic treatment equal to 1/2 mm, and probing pocket depth (PPD) value was decreased in the probiotic bacteria treatment group that can explain the decrease in inflammation in gingiva. Our findings suggest that probiotic mouthwash is healthy for daily use as an alternative for maintaining dental and periodontal health.     

Effects of PM2.5 and NO2 on the 8-isoprostane and lung function indices of FVC and FEV1 in students of Ahvaz city,Iran

The aim of this study was to determine the correlation between PM_2.5 and NO₂ pollutants and oxidative stress marker (8-isoprostane) and lung function tests (FVC and FEV₁) in healthy children who were living and studying in three different areas of Ahvaz city including A₁: Naderi site with high traffic, A₂: Alavi Alley site with average traffic, and A₃: Ein 2 site with low traffic (a rural area on the suburb of Ahvaz). 30 students in the 12–13 year-old range were selected from each studied zone (1, 2 and 3 sites) during three months of year. Of each student, one sample was taken every two weeks to measure 8-isoprostane of exhaled breath condensate (EBC). Air pollution data were collected from three air quality monitoring stations. Also, the relationship between air pollution and 8-isoprostane as well as lung function tests were determined using generalized estimating equations (GEE). The mean concentration of PM_2.5 and NO₂ in A₁, A₂ and A₃ areas were 116, 92 and 45 (μg/m³) also 77, 53 and 14 (ppb) respectively. Among all studied students, there was a significant correlation between the increase of mean concentration of PM_2.5 and NO₂ in 1–4 before sampling day, increased 8-isoprostane concentration and decreased FEV₁, while there was no significant correlation between them and decreased FVC. In A₁ site, an increase in IQR (13 μg/m³) PM_2.5 and IQR (6.5 ppb) NO₂ on 1–4 days before sampling was associated with 0.38 unit (95% CI: 0.11, 0.65) and 1.1 unit (95% CI: 0.85, 1.35) increase in 8-isoprostane concentration, also decreased 121 ml and 190 ml FEV₁, respectively. Results showed that the short-term exposure to traffic-related air pollution can decrease the values of lung function indices and increase the oxidative stress. It may adversely affect children’s lungs.