Live-Attenuated Lentivirus Immunization Modulates Innate Immunity and Inflammation while Protecting Rhesus Macaques from Vaginal Simian Immunodeficiency Virus Challenge

Immunization with attenuated lentiviruses is the only reliable method of protecting rhesus macaques (RM) from vaginal challenge with pathogenic simian immunodeficiency virus (SIV). CD8(+) lymphocyte depletion prior to SIVmac239 vaginal challenge demonstrated that a modest, Gag-specific CD8(+) T cell response induced by immunization with simian-human immunodeficiency virus 89.6 (SHIV89.6) protects RM. Although CD8(+) T cells are required for protection, there is no anamnestic expansion of SIV-specific CD8(+) T cells in any tissues except the vagina after challenge. Further, SHIV immunization increased the number of viral target cells in the vagina and cervix, suggesting that the ratio of target cells to antiviral CD8(+) T cells was not a determinant of protection. We hypothesized that persistent replication of the attenuated vaccine virus modulates inflammatory responses and limits T cell activation and expansion by inducing immunoregulatory T cell populations. We found that attenuated SHIV infection decreased the number of circulating plasmacytoid dendritic cells, suppressed T cell activation, decreased mRNA levels of proinflammatory mediators, and increased mRNA levels of immunoregulatory molecules. Three days after SIV vaginal challenge, SHIV-immunized RM had significantly more T regulatory cells in the vagina than the unimmunized RM. By day 14 postchallenge, immune activation and inflammation were characteristic of unimmunized RM but were minimal in SHIV-immunized RM. Thus, a modest vaccine-induced CD8(+) T cell response in the context of immunoregulatory suppression of T cell activation may protect against vaginal HIV transmission.     

The effects of biochars produced in different pyrolsis temperatures from agricultural wastes on cadmium uptake of tobacco plant

Abiotic stresses caused by cadmium (Cd) contamination in soil retard plant growth and decline the quality of food. Amendment of biochar was reported effective in reduction of mobility, plant uptake and toxicity of Cd in plants. The aim of this study was to investigate the effect of biochar applications produced from corn cob and rice husk at three different pyrolysis temperatures (400, 500 and 600 °C) on Cd uptake of tobacco plants. The results showed that the shoot Cd concentration and content of tobacco plants significantly increased with the application of Cd in increasing doses. The results showed that increasing Cd dosescaused significant increase (P < 0.01) in shoot Cd concentration and content of the tobacco plant at three different pyrolysis temperatures of both corn cob and rice husk biochars. The concentration of Cd was 0.48 mg kg⁻¹ in Cd0 dose of corn cob biochar produced at 500 °C and increased to 61.6 mg kg⁻¹ at Cd5, while Cd concentration increased to 72.3 mg kg⁻¹ with rice husk biochar. Despite the increase in Cd concentrations and content, shoot Cd concentrations and contents were significantly (P < 0.01) reduced with the treatments of corn cob and rice husk biochars produced at different pyrolysis temperatures. The Cd concentration at Cd5 dose in the absence of biochar addition was 90.5 mg kg⁻¹, while Cd concentration at Cd5 dose in 400, 500 and 600 °C treatments of corn cob biochar was reduced to 66.5, 61.6 and 67.3 mg kg⁻¹ respectively, and to 77.0, 72.3 and 70.2 mg kg⁻¹ in rice husk biochar. The results also revealed that corn cob biochar treatments were more effective in reducing Cd uptake of tobacco plants compared to rice husk biochar. Higher specific surface area of corncob biochar compared to rice husk biochar caused to the difference between two biochar sources on Cd uptake of tobacco plants.     

Comparison of amplification refractory mutation system and polymerase chain reaction-restriction fragment length polymorphism techniques used for the investigation of MEFV gene exon 10 point mutations in familial Mediterranean fever patients living in Cukurova region (Turkey)

Familial Mediterranean fever (FMF) is an autosomal recessive inherited disease characterized by recurrent fever, serositis and arthritis. The disease is highly prevalent in Mediterranean basin populations. Recently, the gene responsible for FMF (MEFV) was cloned and at least 40 MEFV gene mutations have been identified. The most frequently observed mutations in the MEFV gene are M694V, M694I, M680I, and V726A. These occur within exon 10 of the gene, and account for 85% of the known MEFV alleles. In this study, the reliability and economical aspects of amplification refractory mutation system (ARMS) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques were compared for analyzing the frequencies of the major point mutations of 90 unrelated patients with FMF from the Cukurova region in Turkey. Both techniques yielded similar results: The ratio of independent alleles of 90 patients carrying one of the tested mutations was 81.1%; patients consisted of 12 different genotypes. In 64 of 90 patients (71.1%) mutations were observed in both alleles. Thirty-six patients (40%) were homozygous for the same mutation, 28 (31.1%) were heterozygous for different mutations. Eighteen patients (20%) were heterozygous for one allele with one of the four mutations but the other allele was unknown. In 8 patients (8.8%) no mutation could be detected. The most frequently observed mutation was M694V (51.66%), followed by M680I (17.22%), V726A (10.55%), and M694I (1.66%). In conclusion ARMS and PCR-RFLP techniques were equally reliable to detect the mutations in Turkish FMF patients. However, the ARMS technique was found to be more rapid and economical than the PCR-RFLP techniques.     

Biology of three species of Agistemus (Acari: Stigmaeidae): life table parameters using eggs of Panonychus citri or pollen of Malephora crocea as food

The biology and life table parameters of Agistemus industani Gonzalez, A. cyprius Gonzalez, and A. floridanus Gonzalez (Acari: Stigmaeidae) were studied under laboratory conditions using two food sources: Panonychus citri (McGregor) eggs or ice plant, Malephora crocea (Jacquin) Schwantes pollen at 25 degrees C. The larval, protonymph, deutonymph, and adult stages of A. industani fed on citrus red mite eggs. All active stages of A. industani, except the larva, fed on all P. citri stages and the larval stage could not feed on P. citri adults. All immature stages of A. industani fed on M. crocea pollen. Agistemus cyprius larvae fed on P. citri eggs and larvae or ice plant pollen. The nymphal stages fed on P. citri eggs, larvae, and protonymphs but not deutonymphs or adults while A. cyprius deutonymphs and adults fed on all P. citri stages. Adult and nymphal stages of A. cyprius fed on ice plant pollen and successfully completed their development while A. floridanus did not. Agistemus floridanus larvae fed only on P. citri eggs, while the other stages fed on P. citri eggs, larvae, and protonymphs. The developmental times from egg to adult for A. industani and A. cyprius when fed M. crocea pollen were 11.3 and 13.4 days, respectively. Agistemus floridanus was unable to complete its life cycle on a diet of only M. crocea pollen. Agistemus industani, A. cyprius, and A. floridanus completed development from egg to adult in 11.7, 13.8, and 10.8 days, respectively, when fed P. citri eggs. The intrinsic rate of increase (r(m)) values for A. cyprius and A. industani were 0.0311 and 0.1201 per day on the pollen diet. The net reproductive rate (Ro) was 3.58 for A. cyprius and 10.07 for A. industani with generation times (T) of 45.2 and 35.1 days, respectively, on the ice plant pollen diet. The r(m) values for A. cyprius, A. floridanus, and A. industani on the P. citri egg only diet were: 0.0562, 0.1001, and 0.1031 per day, respectively. The Ro values for each species fed P. citri eggs only were: 6.36, 7.90, and 18.70 for A. cyprius, A. floridanus, and A. industani and the generation times (T) for each of the three species were: 35.2, 29.9 and 37.8 days, respectively.     

The effect of desferrioxamine on peroxynitrite-induced oxidative damage in erythrocytes

The aim of this study was to investigate the effect of desferrioxamine on peroxynitrite-mediated damage in erythrocytes by measuring the 3-nitrotyrosine level and glutathione peroxidase and Na(+)-K(+) ATPase activities in vitro. 3-Nitrotyrosine levels were determined by HPLC; glutathione peroxidase and Na(+)-K(+) ATPase activities were measured by spectrophotometry. Peroxynitrite increased the 3-nitrotyrosine level but decreased both enzyme activities. In the presence of desferrioxamine, glutathione peroxidase activity was increased with a decrease in the 3-nitrotyrosine level. Desferrioxamine was found to possess an important antioxidant activity as assessed in an in vitro system, reducing protein nitration, restoring enzyme activities and maintaining erythrocyte membrane integrity.     

Risk-Based Zoning Strategy for Soil Remediation at an Industrial Site

After determining at an early stage of the project that the future land use of this New Jersey chemical manufacturing site remain industrial in nature, the site was zoned according to risk. The chemicals of concern (COCs) at the site included relatively low levels of mono- and polynuclear aromatic hydrocarbons, chlorinated aliphatics, as well as other volatile and semivolatile compounds. Direct human exposure scenarios were the key to the mitigation of risks related to soils because the groundwater migration pathway was already interrupted using groundwater recovery. A focused remedial strategy was developed to ensure that the exposure pathways (inhalation, ingestion, and dermal contact) are alleviated and the remedial measures are protective to the workers operating and/or maintaining the site. The risk evaluation process included a preliminary risk assessment (Tier 1) based on a comparison with pertinent soil cleanup criteria, a prioritization analysis to rank zones, chemicals and pathways of concern, and an application of the Risk Based Corrective Action (RBCA) approach (Tier 2) for construction worker exposure scenario. The risk assessment identified selected areas that would benefit from remedial actions. Prioritization Analysis classified the site into five high-priority (comprising 97% of the total health-based risk), three medium-priority (contributing to remaining 2 to 3% of the risk), and adequately protected areas. The boundaries and volumes of affected areas were delineated based on confirmatory soil sampling and statistical analyses. The remedial technologies selected for the site have achieved appropriate reduction in risk to comply with all State regulations and include (in addition to the institutional controls): ??Capping the site where only immobilesemivolatile contaminants are present ??Excavation and on-site treatment of the soils impacted by volatile organic com pounds through ex situ low temperature desorption, or alternative “biopile” treatment and natural attenuation, and ??Excavation and off-site disposal of limited volumes of soils This risk-based, integral approach helped identify the real significance of contamination present at the site and facilitated the development of suitable and adequate remedies. Had not it been for this approach, the mere comparison with soil cleanup criteria would have unnecessarily resulted in denoting all areas as nuisance contributors, and thus requiring some actions. New Jersey Department of Environmental Protection (NJDEP) has approved this approach and contributed to its accomplishment.     

Investigation of New Microstrip Bandpass Filter Based on Patch Resonator with Geometrical Fractal Slot

A compact dual-mode microstrip bandpass filter using geometrical slot is presented in this paper. The adopted geometrical slot is based on first iteration of Cantor square fractal curve. This filter has the benefits of possessing narrower and sharper frequency responses as compared to microstrip filters that use single mode resonators and traditional dual-mode square patch resonators. The filter has been modeled and demonstrated by Microwave Office EM simulator designed at a resonant frequency of 2 GHz using a substrate of ε_r = 10.8 and thickness of h = 1.27 mm. The output simulated results of the proposed filter exhibit 22 dB return loss, 0.1678 dB insertion loss and 12 MHz bandwidth in the passband region. In addition to the narrow band gained, miniaturization properties as well as weakened spurious frequency responses and blocked second harmonic frequency in out of band regions have been acquired. Filter parameters including insertion loss, return loss, bandwidth, coupling coefficient and external quality factor have been compared with different values of perturbation dimension (d). Also, a full comparative study of this filter as compared with traditional square patch filter has been considered.     

Natural history of 11 cases of twin-twin transfusion syndrome without intervention.

The natural history of 11 cases of twin-twin transfusion syndrome (TTTS) in monochorionic diamniotic (MCDA) twin pregnancies has been reviewed. Seven cases before 28 weeks and four pregnancies after 28 weeks had been followed up without intervention. Eight cases had premature uterine contractions. All seven pregnancies before 28 weeks aborted, leading to a 100% mortality rate. After 28 weeks all mothers delivered live births. The diagnosis of TTTS before 28 weeks, and with premature uterine contraction, seems to be a poor prognostic sign.     

Insights into subunit interactions in the heterotetrameric structure of potato ADP-glucose pyrophosphorylase

ADP-glucose pyrophosphorylase, a key allosteric enzyme involved in higher plant starch biosynthesis, is composed of pairs of large (LS) and small subunits (SS). Current evidence indicates that the two subunit types play distinct roles in enzyme function. The LS is involved in mainly allosteric regulation through its interaction with the catalytic SS. Recently the crystal structure of the SS homotetramer has been solved, but no crystal structure of the native heterotetrameric enzyme is currently available. In this study, we first modeled the three-dimensional structure of the LS to construct the heterotetrameric enzyme. Because the enzyme has a 2-fold symmetry, six different dimeric (either up-down or side-by-side) interactions were possible. Molecular dynamics simulations were carried out for each of these possible dimers. Trajectories obtained from molecular dynamics simulations of each dimer were then analyzed by the molecular mechanics/Poisson-Boltzmann surface area method to identify the most favorable dimers, one for up-down and the other for side-by-side. Computational results combined with site directed mutagenesis and yeast two hybrid experiments suggested that the most favorable heterotetramer is formed by LS-SS (side-by-side), and LS-SS (up-down). We further determined the order of assembly during the heterotetrameric structure formation. First, side-by-side LS-SS dimers form followed by the up-down tetramerization based on the relative binding free energies.