Human immunodeficiency virus type-1 group M quasispecies evolution: diversity and divergence in patients co-infected with active tuberculosis

The evolution of intra-host human immunodeficiency virus type 1 (HIV-1) quasispecies prior and after treating active tuberculosis (TB) with chemotherapy in HIV-1/TB patients was assessed. Two time points HIV-1 quasispecies were evaluated by comparing HIV-1-infected patients with active tuberculosis (HIV-1/TB) and HIV-1-infected patients without tuberculosis (HIV-1/non-TB). Plasma samples were obtained from the Frankfurt HIV cohort, and HIV-1 RNA was isolated. C2V5 env was amplified by PCR and molecular cloning was performed. Eight to twenty-five clones were sequenced from each patient. Various phylogenetic analyses were performed. We found a significant increase in diversity and divergence in HIV-1/TB compared to the HIV-1/non-TB. For HIV-1/TB, the average rate of evolution of C2V5 env was higher than previous reports (2.4 × 10⁻⁴ substitution/site/day). Two groups of HIV-1/TB were observed based on the rate of HIV-1 evolution and coreceptor usage: A fast evolving R5-tropic dominating group and a relatively slowly evolving X4 group. The results demonstrated that active TB has an impact on HIV-1 viral diversity and divergence over time. The influence of active TB on longitudinal evolution of HIV-1 may be predominant for R5 viruses.     

Hematological assessment of health status of African catfish Clarias gariepinus (Burchell 1822) experimentally challenged with Escherichia coli and Vibrio fischeri

Clarias gariepinus were challenged with one of the following treatments: 1?×?10⁵ colony-forming units (CFU)/ml of Escherichia coli (EC1), 2?×?10⁵ CFU/ml of E. coli (EC2), 1?×?10⁵ CFU/ml of Vibrio fischeri (V1), 2?×?10⁵ CFU/ml of V. fischeri (V2), gavaged with distilled water (CW), and not gavaged (C). Fish inoculation with bacteria was done via oral gavage. Fish were maintained in the laboratory for 7 days after the bacterial inoculation, after which various hematological parameters such as percentage packed cell volume (PCV), percentage red blood cell lysis, white blood cell (WBC) counts, and differential white blood cell counts (neutrophils, lymphocytes, monocytes, and eosinophils) were determined. Fish challenged with bacteria have a significantly reduced percentage PCV but a significantly higher percentage of red blood cell lysis, WBC counts, neutrophils, lymphocytes, monocytes, and eosinophils compared to the controls. In all cases, effects tend to be strongly dependent on the CFU of the bacteria with effects generally more pronounced in fish infected with 2?×?10⁵ CFU/ml of either E. coli or V. fischeri. The results of this study indicate that the health status of C. gariepinus is seriously compromised by bacterial infection, and hematological parameters are reliable enough in the assessment and quick diagnosis of the health status of fish infected with bacteria.     

A Phase 1 Trial of CNDO-109–Activated Natural Killer Cells in Patients with High-Risk Acute Myeloid Leukemia

Natural killer (NK) cells are an emerging immunotherapy approach to acute myeloid leukemia (AML); however, the optimal approach to activate NK cells before adoptive transfer remains unclear. Human NK cells that are primed with the CTV-1 leukemia cell line lysate CNDO-109 exhibit enhanced cytotoxicity against NK cell–resistant cell lines. To translate this finding to the clinic, CNDO-109–activated NK cells (CNDO-109-NK cells) isolated from related HLA-haploidentical donors were evaluated in a phase 1 dose-escalation trial at doses of 3?×?10⁵ (n?=?3), 1?×?10⁶ (n?=?3), and 3?×?10⁶ (n?=?6) cells/kg in patients with AML in first complete remission (CR1) at high risk for recurrence. Before CNDO-109-NK cell administration, patients were treated with lymphodepleting fludarabine/cyclophosphamide. CNDO-109-NK cells were well tolerated, and no dose-limiting toxicities were observed at the highest tested dose. The median relapse-free survival (RFS) by dose level was 105 (3?×?10⁵), 156 (1?×?10⁶), and 337 (3?×?10⁶) days. Two patients remained relapse-free in post-trial follow-up, with RFS durations exceeding 42.5 months. Donor NK cell microchimerism was detected on day 7 in 10 of 12 patients, with 3 patients having evidence of donor cells on day 14 or later. This trial establishes that CNDO-109-NK cells generated from related HLA haploidentical donors, cryopreserved, and then safely administered to AML patients with transient persistence without exogenous cytokine support. Three durable complete remissions of 32.6 to 47.6+?months were observed, suggesting additional clinical investigation of CNDO-109-NK cells for patients with myeloid malignancies, alone or in combination with additional immunotherapy strategies, is warranted.     

Human cell-mediated immunity to tuberculin as assayed by the agarose micro-droplet leukocyte migration inhibition technique: Comparison with the capillary tube assay

Direct and indirect agarose microdroplet migration inhibition assays were performed with log dilutions (50?5 × 10^?8 μg/ml) of soluble purified protein derivative (PPD) of tuberculin and leukocytes (4 × 10⁵/droplet) from PPD skin test positive or negative individuals. Some tests were run in parallel with the capillary tube method. Inhibition of migration of leukocytes from 9/11 PPD skin test positive donors studied was observed in direct tests when employing PPD doses ranging from 1–50 μg/ml PPD. Inhibition of migration of leukocytes from some PPD skin reactive donors was obtained with as little as 5 × 10^?3? 5 × 10^?7 μg/ml (i.e., 5 nanograms to 0.5 picograms). Some inhibition of leukocyte migration was seen with skin test negative donors (presumably toxicity) with the higher doses of PPD (50 μg/ml), but generally little inhibition was observed with these donors at lower doses. Comparative experiments of the agarose micromethod and the capillary tube technique indicated that the agarose assay was more sensitive in that it could detect reactivity with 2–4 logs lower antigen concentration. Indirect assays using supernatants of cultures of PPD triggered mononuclear cells and indicator granulocytes in agarose droplets demonstrating that a lymphokine (presumably leukocyte inhibitory factor) was being produced and suggested that cell-mediated immune (CMI) reactions were operative. The results indicate the usefulness of this technique in the sensitive detection of CMI against such antigens as soluble PPD. The assay should prove useful in quantitative assessment of cell-mediated reactivity by using a wide range of antigen concentrations and the leukocytes from as little as 2–5 ml of blood.     

Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. VIII. Adult and fetal guinea pig (Cavia procellus)

The diffusional water permeability (P _d) of adult, pregnant female and fetal guinea-pig red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition withp-chloromercuribenzene sulphonate (PCMBS). The values ofP _d were around 5.0 × 10^?3 cm/s at 15 °C, 5.3 × 10-3 cm/s at 20 °C, 6.6 × 10^?3 cm/s at 25 °C, 7.5 × 10^?3 cm/s at 30 °C and 8.6 × 10^?3 cm/s at 37 °C with no significant differences between adult, pregnant female and fetal RBCs. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 10 min at 37 °C with 0.1 mm PCMBS. The values of maximal inhibition ranged from 70%–77% at 15–30 °C to 57%–63% at 37 °C in the case of adult and from 64%–67% at 15–30 °C to 51% at 37 °C in the case of fetal RBCs. The basal permeability to water was estimated at 1.1 × 10^?3 cm/s at 15 °C ,1.3 × 10^?3 cm/s at 20 °C, 1.6 × 10^?3 cm/s at 25 °C, 2.2 × 10^?3 cm/s at 30 °C and 3.2 × 10^?3 cm/s at 37 °C for adult and slightly higher values for fetal guinea pig RBCs as 1.6 × 10^?3 cm/s at 15 °C, 2.0 × 10^?3 cm/s at 20 °C, 2.4 × 10^?3 cm/s at 25 °C, 2.6 × 10^?3 cm/s at 30 °C and 4.2 × 10^?3 cm/s at 37 °C. The activation energy of water diffusion was around 22 kJ/ mol, with no significant differences between the adult pregnant female and fetal RBCs, and increased to about 40 kJ/mol in the case of adult and pregnant RBCs and 34 kJ/mol for fetal RBCs after incubation with PCMBS in conditions of maximal inhibition of water diffusion. The membrane polypeptide electrophoretic pattern of adult and fetal guinea-pig RBCs was compared with its human counterpart. The guinea-pig membrane contained higher amounts of spectrin (band 1 and 2), whereas the proteins in bands 4.1, 4.2 and 6 were present in lower amounts. Considerable differences in polypeptides migrating in the region of bands 7 and 8 and in front of them were apparent between the two sources of RBC membranes where some bands were present only in the guinea-pig RBC membranes. The adult guinea-pig membranes contained smaller amounts of proteins migrating in band 4.5 and lacked band 8.     

Molecular genetic analysis and evolution of begomoviruses and betasatellites causing yellow mosaic disease of bhendi

In India, Bhendi yellow vein mosaic disease (BYVMD) is one of the most economically important diseases of bhendi/okra and is caused by a complex of monopartite begomovirus (Bhendi yellow vein mosaic virus—BYVMV) and betasatellite (Bhendi yellow vein betasatellite—BYVB). In this study, we have analyzed the role of possible evolutionary factors involved in the evolution of BYVMV and BYVB isolates. Evidence of inter-species and inter-strain recombination events was detected among the viral isolates, and majority of these recombinant isolates possess microsatellites in their genome. Recombination analysis suggests that cotton-infecting and bhendi-infecting begomoviruses probably share a recent common ancestor. In addition to genetic differentiation and gene flow, high degree of genetic variability was detected among the viral population. A strong purifying selection seems to be acting on the viral coding regions. The nucleotide substitution rate of V1 gene (for BYVMV) and βC1 gene (for BYVB) was estimated to be 7.55 × 10^?4 and 2.25 × 10^?3 nucleotide substitutions/site/year, respectively. The present study underlines that the evolution of BYVMD-associated viral components is driven by selection acting on the genetic variation generated by recombination and mutation.     

Genetic characterization of bovine coronavirus in Vietnam

A maximum clade credibility tree constructed using the full-length spike (S) and hemagglutinin-esterase genes revealed that Vietnamese Bovine coronavirus (BCoV) strains belong to a single cluster (C1); therefore, they might share a common origin with Cuban and Chinese BCoV strains. The omega values of cluster 1 (C1) and cluster 2 (C2) were 0.15734 and 0.11613, respectively, and naive empirical bayes analysis identified two amino acid positions (179 and 501) in the S protein in C1 and three amino acid positions (113, 501, and 525) in that of C2 that underwent positive selection (p?>?99%). The evolutionary rate of C1 was estimated to be 7.6206?×?10^?4 substitutions/site/year, and the most recent common ancestor (tMRCA) of Vietnamese BCoVs was estimated to date back to 1962 (95% HPD 1950–1973). The effective population sizes of C1 and C2 underwent a rapid reduction after 2000 and 2004, respectively.

     

Comparative NMR studies of diffusional water permeability of red blood cells from different species. X. Camel (Camelus dromedarius) and alpaca (Lama pacos)

The diffusional water permeability (P _d) of camel and alpaca red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition withp-chloromercuribenzene sulphonate (PCMBS). The values ofP _d were, in the case of alpaca RBC≈4.6×10^?3 cm/s at 25°C, 5.4×10^?3 cm/s at 30°C, 6.6×10^?3 cm/s at 37°C and 7.7×10^?3 cm/s at 42°C. In case of camel RBC the values ofP _d where ≈4.2×10^?3 cm/s and 9.0×10^?3 cm/s at 42°C. Systematic studies on the effects of PCMBS on water diffusion in camel RBC indicated that the maximal inhibition was reached in 45 min with 1–2 mm PCMBS. The values of maximal inhibition were around 47% at 25°C and 68% at 30°C for alpaca RBC and around 62% at 25°C and 56% at 37°C for camel RBC. The basal permeability to water of alpaca RBC was estimated at around 2.6×10^?3 cm/s at 25°C, 1.7×10^?3 cm/s at 30°C and of camel RBC as 1.8×10^?3 cm/s at 25°C and 3.0×10^?3 cm/s at 37°C. The values of the activation energy of water diffusion (E _{a, d}) were around 23 kJ/mol for camel and 34 kJ/mol for alpaca RBC. This suggests that in addition to the number of transport channels other features of the pathways might be important for defining the temperature dependence of the water permeability.     

Configurational double bond selectivity in the epoxidation of hydroxy-terminated polybutadiene with m-chloroperbenzoic acid

The selectivity of the three different double bonds (CIS, TRANS and VINYL) of hydroxyterminated polybutadiene regarding epoxidation was evaluated, using m-chloroperbenzoic acid in toluene below room temperature (?10°C, ?5°C, 0°C and 5°C). To determine the kinetic constants for the three configuratons (k_1,cis = (2,59–0,6) × 10^?2 L · mol^?1 · min^?1; k_{2, trans} = (5,35–0,82) × 10^?2 L · mol^?1 · min^?1; k_{3, vinyl} = (0,97–0,01) × 10^?2 L · mol^?1 min^?1), ¹H and ¹³C NMR was used along with a system of three parallel equations. The activation energy values were also evaluated (E_{a, cis} = 53,9 kJ · mol^?1; E_{a, trans} = 70,7 kJ · mol^?1 and E_a, vinyl = 261,1 kJ · mol^?1) for the three double bond types.     

Association of genetic variants in estrogen receptor α with HCV infection susceptibility and viral clearance in a high-risk Chinese population

The purpose of this study was to test the hypothesis that genetic variants of estrogen receptor α (ERα) are associated with the outcomes of hepatitis C virus (HCV) infection. We genotyped the seven single nucleotide polymorphisms (SNPs) (rs2077647, rs9340799, rs2234693, rs1801132, rs9322354, rs2228480 and rs3798577) of ERα and conducted a case-control study in a high-risk Chinese population, including 429 HCV spontaneous clearance cases, 880 persistent infection cases and 1,174 uninfected controls. The C allele of rs2234693 was significantly associated with increased susceptibility to HCV infection [dominant model: adjusted odds ratio (OR)?=?1.377, 95 % confidence interval (CI) =1.126–1.778], and the risk effect remained significant among the younger (≤55 years) and hemodialysis subjects (all P?<?0.007). The other three SNPs variant genotypes also showed significant correlation with elevated risk of HCV infection in different strata (rs2077647 in males; rs9340799 in blood donors; rs1801132 in younger subjects; all P?<?0.007). It was also discovered that carriage of rs2228480 A allele was more prone to develop persistent HCV infection (dominant model: adjusted OR?=?1.203, 95 % CI?=?1.154–1.552), and the risk effect was more evident in females and blood donors (all P?<?0.007). Haplotype analyses (rs2077647, rs9340799 and rs2234693) showed that, compared with the most frequent haplotype TAT, CAC played a risk effect in subgroups of younger (P?=?3.24?×?10^?3) and male (P?=?5.51?×?10^?4), whereas CAT expressed a protective effect in females (P?=?2.27?×?10^?4) for HCV infection susceptibility. We first report that these SNPs (rs2077647, rs9340799, rs2234693, rs1801132 and rs2228480) in ERα can influence the outcomes of HCV infection in a high-risk Chinese population.     

Haematological reference ranges of cultured Clarias gariepinus in the Lower Benue River Basin,Nigeria

Blood samples were collected from 170 cultured African sharptooth catfish (Clarias gariepinus) to establish haematological baseline values of this important tropical pisciculture fish species in the Guinea savannah agro-ecological zone of Nigeria. The total red blood cell count and the total white blood cell count were obtained by haemocytometry, while the packed cell volume and haemoglobin were obtained by microhaematocrit and cyanomethmoglobin methods, respectively. The results obtained varied between gender and age and were as follows: total red blood cell count, 1.72?±?0.34?×?10⁶/mm³ in juveniles and 4.50?±?0.57?×?10⁶/mm³ in adults; total white blood cell count, 15.50?±?1.1510³/mm³in juvenile and 16.41?±?1.21?×?10³/mm³in adults; packed cell volume, 30.08?±?7.78 % in juveniles and 39.59?±?3.93 % in adults; haemoglobin, 9.43?±?3.45 g/dl in juveniles and 10.99?±?3.29 g/dl in the adults; mean corpuscular volume, 173.75?±?41.93 fl in juveniles and 87.01?±?14.37 fl in adults; mean corpuscular haemoglobin, 51.11?±?13.10 pg in the juveniles and 26.81?±?8.61 pg in the adults, while the mean corpuscular haemoglobin concentration values obtained are 32.61?±?10.42 g/dl in the juveniles and 33.80?±?10.0 g/dl in the adults.     

Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples

In this study, a quantitative real-time PCR was developed to detect and quantify Acanthamoeba spp. in various environmental water samples. The water samples were taken from watershed, water treatment plant, and three thermal spring recreation areas. The overall detection rate was 14.2 % (25/176) for Acanthamoeba spp. The percentages of samples containing Acanthamoeba spp. from river water, raw drinking water, and thermal spring water were 13 % (13/100), 25 % (7/28), and 10.4 % (5/48), respectively. Acanthamoeba spp. concentrations were determined according to SYBR Green quantitative real-time PCR. A plasmid-based standard curve was constructed to determine the Acanthamoeba concentration using dilution factors for achieving 1.36?×?10⁹ gene copies per PCR for 18S rRNA gene in Acanthamoeba spp. The resulting concentrations varied by the type of water, in the range of 46–2.6?×?10² cells/l in positive raw drinking water, 2.7?×?10²–1.5?×?10⁴ cells/l in river water, and 54–1.7?×?10³ cells/l in thermal spring water. The presence of Acanthamoeba spp. in the raw drinking water samples was also found to have a significant difference with heterotrophic plate count. The presence of Acanthamoeba spp. in various aquatic environments may be a potential health hazard and must be further evaluated.     

Hematology of the Mediterranean population of sea turtle (Caretta caretta): comparison of blood values in wild and captive, juvenile and adult animals

In order to establish baseline hematological and biochemical values in loggerhead turtles from the Mediterranean Sea, 84 specimens were sampled, comprising 24 wild turtles in good health at the time of capture and 60 turtles tested after indoor rehabilitation at the Sea Turtle Rescue and Rehabilitation Centre of the Zoological Station Anton Dohrn in Naples, Italy. The following parameters were evaluated: red cell counts (RBC, 488–575?×?10³/μL), white cell counts (WBC, 17–24?×?10³/μL) and thrombocyte counts (TBC, 19–49?×?10²/μL), hemoglobin (Hb, 8–14?g/dL), hematocrit (Ht, 23–34%), mean corpuscular volume (MCV, 487–723?fL), mean corpuscular hemoglobin (MCH, 170–261?pg), mean corpuscular hemoglobin concentration (MCHC, 34–42%), white and red blood cell differential counts, and a panel of hematochemical tests, composed of glucose (97–164?mg/dL), cholesterol (74–144?mg/dL), blood urea nitrogen (35–200?mg/dL), uric acid (1–2.7?mg/dL), total bilirubin (0.20–0.40?mg/dL), GOT (44–184?IU/L) and GPT (6?IU/L) transaminases, calcium (6.7–8.7?mg/dL), and magnesium (3.6–5.4?mEq/L). Comparisons of the statistically analyzed data from the turtles which were divided into groups on the basis of age and/or lifestyle (wild or captive) revealed that erythroid parameters attained higher values in captive turtles. This suggested a positive influence of the rich and complete diet fed in captivity upon the hemopoietic process of the turtle. On the other hand, data suggest a more intense and active hemopoiesis in young turtles, compared to adult specimens.     

Impact of Single-Dose Plerixafor as an Adjunct to Granulocyte Colony-Stimulating Factor–Based Peripheral Blood Stem Cell Mobilization on the Graft Composition and Outcome for T Cell–Replete Haploidentical Peripheral Blood Stem Cell Transplantation with Post-Transplantation Cyclophosphamide: A Comparative Study

We conducted a prospective study on T and natural killer (NK) cell subset composition of graft and transplant outcomes in T cell–replete haploidentical transplantation with a single dose of subcutaneous plerixafor (Px) added to granulocyte colony-stimulating factor (G-CSF)-based mobilization in allogeneic donors to collect 10?×?10⁶/kg CD34⁺ hematopoietic stem cells (HSCs) at single apheresis. Twnety-six donors received G-CSF?+?Px and 25 G-CSF alone for mobilization. Despite significantly lower peripheral blood (PB) CD34⁺ HSCs on day 4 in the G-CSF?+?Px group (33 [range, 6-47] cells/µL versus 81 [range, 50-168] cells/µL in the G-CSF group; P?=?.0001), PB CD34⁺ HSC count (median 136 versus 139 cells/µL) on day 5 as well as that in the graft (2.7 versus 2.3?×?10⁶/mL, P?=?.1) were comparable between the 2 groups. The total nucleated cell count was higher (3.4 versus 3.1?×?10⁸/mL, P?=?.05), but CD4⁺ T cells (2.3 versus 2.7?×?10⁷/mL, P?=?.09) were lower in the G-CSF group with mobilization of regulatory T cells being similar. NK cells were skewed toward the CD56⁺/16^? subset in both groups, varying significantly from the steady-state NK subset ratio in PB. The time to engraftment, incidences of acute and chronic graft-versus-host disease, nonrelapse mortality, and overall survival were also similar. Addition of single-dose Px to G-CSF mobilization improves CD34 recovery and does not significantly alter the T and NK cell composition of the graft, including regulatory T cells, with no adverse impact on transplant outcomes.     

Nucleotide sequence of the gene encoding the RNA polymerase and the 3′ non-coding region of a bovine enterovirus Japanese isolate: rapid synonymous substitutions between European and Japanese strains

Summary.  The nucleotide sequences of the genome RNA encoding the RNA polymerase and the 3′ non-coding region (NCR) of bovine enterovirus (BEV) serotype I Japanese isolate, MZ468, were determined. The genetic distance between the two BEV serotype I strains, MZ468 and VG-5-27, was calculated by pairwise comparison of nucleotide sequences. The synonymous substitution rate was high (1.40 × 10⁻²/site/year), and of the same order as those of influenza virus HA, HIV-1 gag and env, and enterovirus 70 VP1 genes. Accepted December 1, 1997 Received June 2, 1997     

Evolution rate of hepatitis delta virus RNA isolated in Taiwan

The complete RNA sequences of hepatitis delta viruses (HDV) isolated at 3 years apart from a chronic delta hepatitis patient in Taiwan were determined. The sequence analysis showed an overall evolution rate of 3.18 × 10^?3 substitutions/nucleotide/year. The evolution rates in different parts of HDV RNA varied. The hypervariable region evolved faster (4.55 × 10^?3 substi-tutions/nucleotide/year) than the hepatitis delta antigen (HDAg)-coding region (2.60 × 10^?3 substitutions/nucleotide/year) and the autocata-lytic region (1.11 × 10^?3 substitutions/nucleotide/ year). These data are compatible with the previous finding that the hypervariable region is more divergent than the HDAg-coding region and the au-tocatalytic regions among the HDV isolates from different geographic areas. No substitution was found in the four previously identified conserved domains of HDV RNA, further confirming their functional importance in viral replication. The evolution rate of this HDV RNA is higher than that determined from the partial RNA sequences of two Japanese HDV isolates and similar to that found in a Lebanon isolate. Further, it was found that this HDV RNA retained the same microheterogeneities at 15 nucleotide positions detected in the RNA 3 years earlier. It is concluded that HDV RNA in patients' serum is extremely heterogeneous, and that the nucleotide substitutions in certain nucleotide positions likely have conferred evolutionary advantages for HDV. Viral sequence evolution is a possible mechanism for chronic HDV infection. © 1994 Wiley-Liss, Inc.     

The degree of HIV-1 amino acid variability is strictly related to different disease progression rates

The aim of this study is to evaluate the amino acid variability of HIV-1 Gp41, C2–V3, and Nef in a group of patients characterized by different disease progression rates. HIV-1 sequences were collected from 19 Long term non progressor patients (LTNPs), 9 slow progressors (SPs), and 11 rapid progressors (RPs). Phylogenetic trees were estimated by MEGA 6. Differences in amino acid variability among sequences belonging to the 3 groups have been evaluated by amino acid divergence, Shannon entropy analysis, and the number of amino acid mutations (defined as amino acid variations compared with HxB2). The involvement of amino acid mutations on epitope rich regions was also investigated. The population was mainly composed of males (74.3%) and HIV-1 subtype B strains (B: 92.32%, CRF_12BF, A1, C: 2.56% each). Viral load (log₁₀ copies/mL) and CD4⁺T cell count (cells/mm³) were 3.9 (3.5–4.2) and 618 (504–857) in LTNPs, 3.3 (2.8–4.7) and 463 (333–627) in SPs, and 4.6 (4.3–5.3) and 201 (110–254) in RPs. Gp41 and C2–V3 amino acid divergence was lower in LTNP and SP strains compared to RPs (median value: 0.085 and 0.091 vs. 0.114, p?=?0.005 and 0.042) and a trend of lower variability was observed for Nef (p?=?0.198). A lower entropy value was observed at 10, 3, and 7 positions of Gp41, C2–V3, and Nef belonging to LTNPs and at 7, 3, and 1 positions of Gp41, C2–V3, and Nef belonging to SPs compared with RPs (p?<?0.05). Focusing on epitope rich regions, again a higher degree of conservation was observed in Gp41 and C2–V3 sequences belonging to LTNPs and SPs compared to those belonging to RPs. This study shows that the extent of amino acid variability correlates with a different HIV-1 progression rate. This variability also involves CTL epitope rich regions, thus suggesting its involvement in the immune escape process modulation.     

Sterilization of Escherichia coli O157:H7 using micro corona ionizer

We demonstrated in vitro sterilization of Escherichia coli O157:H7 bacteria on agar by a pin-between-planes micro corona ionizer. The gap between the pin and the grid was ~1.1 mm, the length of the grid was ~2.1 mm and the height was ~1.0 mm. The effective pin radius and discharge length were both approximated to be 200 μm. Ozone generation rates of ~2.3?×?10^?3 mg/s, ~2.7?×?10^?3 mg/s and ~3.5?×?10^?3 mg/s at 1,500 V were calculated for relative humidity (RH) of 35 %, 25 % and 10 % respectively. Analytical ozone generation rate increases as RH decreases and it is consistent with experimental observations. Using target and control petri dishes with E. coli plated agar, the sterilization capability of the micro corona ionizer at 37 °C for 24 h was evaluated. A ~60 % reduction in bacterial colony was shown with plate counting and its kill radius could be tuned from?~?20 mm to ~5 mm by reducing the duty cycle from 100 % to 50 % with 30 min pulse width. The results suggested that the micro corona ionizer might be suitable as a tunable ozone source in wound dressing for chronic wound management.     

Electromyography as a new means of navigation during endotracheal intubation

Abstract

This study tested a method of using rapid analysis of electromyographic response patterns to electrical stimulation to enable real-time navigation during endotracheal intubation. An electromyographic response detection device was constructed and integrated into a standard endotracheal tube. The rebound rates of the response voltages were measured in the trachea and oesophagus after stimulation in an acute study performed in three freshly euthanized male Suffolk sheep. In a blind study, a physician attempted to identify the tissue type solely from the electrical response signals. In the acute study, the observed rebound rate was found to be significantly faster in tracheal tissue (2.21?×?10^?3 V s^?1) than in oesophageal tissue (3.45?×?10^?2 V s^?1; p?=?0.000 05). In the blind study, the physician correctly determined the oesophagus response rate seven out of eight times and the tracheal rate eight out of nine times. These results suggest that electromyographic responses can be used to accurately differentiate tracheal from oesophageal tissue during ETT insertion, thus offering a valuable new means of enhancing patient safety.