Effect of providing citrus pulp-integrated diet on fecal microbiota and serum and fecal metabolome shifts in crossbred pigs.

The study aimed to assess the impact of dehydrated citrus pulp (DCP) on growth performance, fecal characteristics, fecal bacterial composition (based on 16S rRNA analysis), and fecal and serum metabolomic profiles in crossbred pigs. 80 finishing pigs Duroc × (Landrace × Large White) were fed either a control diet (C) or a diet with 240 g/kg DCP (T) for six weeks. Including DCP in diets tended to decrease feed intake, increased (p < 0.05) the concentrations of acetic and heptanoic acids and decreased (p < 0.05) fecal butyric and branched-chain fatty acid concentrations in feces. Animals fed DCP exhibited a lower abundance of the genera Clostridium and Romboutsia, while Lachnospira significantly increased. Orthogonal partial least squares discriminant analysis plotted a clear separation of fecal and serum metabolites between groups. The main discriminant fecal metabolites were associated with bacterial protein fermentation and were downregulated in T-fed pigs. In serum, DCP supplementation upregulated metabolites related to protein and fatty acids metabolism. In conclusion, the addition of DCP as an environmentally friendly source of nutrients in pig diets, resulted in modifications of fecal bacterial composition, fermentation patterns, and overall pig metabolism, suggesting improvements in protein metabolism and gut health.

Preliminary report of transmitted drug resistance to integrase strand chain transfer inhibitors in treatment-naïve HIV infected patients.

Transmitted Resistance exists in a newly diagnosed person who has not yet started their treatment. Our objective was to obtain a profile of HIV-1 resistance to integrase inhibitors in newly diagnosed treatment-naïve patients. Fifty people newly diagnosed with HIV-1 infection who had never received antiretroviral treatment were recruited. The complete integrase gene was amplified by nested RTPCR and the sequences obtained were analyzed with the ReCall and HIVdb v9.0. The overall prevalence transmitted due to mutations with some impact on integrase strand transfer inhibitors (INSTI) activity during the study period was 8%. The major E138K mutation was detected in only 1 patient and the secondary G163R mutation was detected in the other 3. The transmitted resistance for the first generation INSTI was 8% and for the second generation it was 0%. In Chile the resistance transmitted to INSTI is low and it is in according values detect in other part of the world.

Sustained neurotrophin-3 delivery from hyaluronic acid hydrogels for neural tissue regeneration.

The goal of this work was to design a polymer-based platform capable of localized, long-term delivery of biologically active neurotropic factors using an affinity-based approach. Here, we synthesized hyaluronic acid-methylfuran (HA-mF) hydrogels that provide sustained, affinity-based release of neurotrophin-3 (NT-3), a growth factor that promotes axon growth for 28 days. A Diels-Alder crosslinking reaction between HA-mF and polyethylene glycol (PEG)-dimaleimide occurs within 15 min under physiological conditions, resulting in hydrogels that can be polymerized in the presence of cells and growth factors. We also tuned the hydrogel's storage modulus to match that of native rat spinal cord tissue, providing a platform not only for localized drug delivery but also a suitable vehicle for cellular transplantation. The NT-3 released from the HAmF hydrogels remains bioactive for at least 14 days, promoting axonal growth from primary sensory neurons as well as stem cell-derived V2a interneurons and motoneurons in vitro. The hydrogels also supported cell growth allowing for 3-dimensional axonal extensions within the scaffold matrix. Here we confirm the protective role of HA-mF on matrix-bound NT-3 activity and show that these hydrogels are an excellent platform for growth factor delivery for neural applications.

Microbial and immune faecal determinants in infants hospitalized with COVID-19 reflect bifidobacterial dysbiosis and immature intestinal immunity.

The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread rapidly worldwide, seriously endangering human health. Although SARS-CoV-2 had a lower impact on paediatric population, children with COVID-19 have been reported as suffering from gastrointestinal (GI) symptoms at a higher rate than adults. The aim of this work was to evaluate faeces as a source of potential biomarkers of severity in the paediatric population, with an emphasis on intestinal microbiota and faecal immune mediators, trying to identify possible dysbiosis and immune intestinal dysfunction associated with the risk of hospitalization. This study involved 19 patients with COVID-19 under 24 months of age hospitalized during the pandemic at 6 different hospitals in Spain, and it included a comparable age-matched healthy control group (n = 18). Patients and controls were stratified according to their age in two groups: newborns or young infants (from 0 to 3 months old) and toddlers (infants from 6 to 24 months old). To characterize microbial intestinal communities, sequencing with Illumina technology of total 16S rDNA amplicons and internal transcribed spacer (ITS) amplicons of bifidobacteria were used. Faecal calprotectin (FC) and a range of human cytokines, chemokines, and growth factors were measured in faecal samples using ELISA and a multiplex system. Significant reduction in the abundance of sequences belonging to the phylum Actinobacteria was found in those infants with COVID-19, as well as in the Bifidobacteriaceae family. A different pattern of bifidobacteria was observed in patients, mainly represented by lower percentages of Bifidobacterium breve, as compared with controls. In the group of hospitalized young infants, FC was almost absent compared to age-matched healthy controls. A lower prevalence in faecal excretion of immune factors in these infected patients was also observed. CONCLUSION:  Hospitalized infants with COVID-19 were depleted in some gut bacteria, such as bifidobacteria, in particular Bifidobacterium breve, which is crucial for the proper establishment of a functional intestinal microbiota, and important for the development of a competent immune system. Our results point to a possible immature immune system at intestine level in young infants infected by SARS-CoV2 requiring hospitalization. WHAT IS KNOWN: • Although SARS-CoV-2 had a lower impact on paediatric population, children with COVID-19 have been reported as suffering from gastrointestinal symptoms at a higher rate than adults. • Changes in microbial composition have been described in COVID-19 adult patients, although studies in children are limited. WHAT IS NEW: • The first evidence that hospitalized infants with COVID-19 during the pandemic had a depletion in bifidobacteria, particularly in Bifidobacterium breve, beneficial gut bacteria in infancy that are crucial for the proper establishment of a competent immune system. • In young infants (under 3 months of age) hospitalized with SARS-CoV2 infection, the aberrant bifidobacterial profile appears to overlap with a poor intestinal immune development as seen by calprotectin and the trend of immunological factors excreted in faeces.

The Duration of the Trial Influences the Effects of Mineral Deficiency and the Effective Phytase Dose in Broilers' Diets.

Two trials varying in duration (short- and long-term) were conducted to evaluate the effects of providing deficient (NC) or sufficient (PC) Ca and P levels, and different doses of a new phytase (250, 500, and 1000 FTU/kg feed), in broiler feed on growth performance, nutrient digestibility and retention, and tibia mineralization. A total of 80 and 490 male chicks (Ross) of 21 and 1 days of age were used in the short- and long-term trials, respectively. In the long-term trial, chicks fed NC diets showed a lower (p < 0.05) average daily gain and feed intake compared to chicks fed PC and a greater (p < 0.05) feed conversion ratio compared to 500 and 1000 FTU/kg feed during the starting period. Regarding the effects on minerals' and nutrients' coefficients of retention, animals fed NC showed a significantly higher digestibility for P than those fed the PC diet in the long-term trial. Additionally, feeding 250 to 500 FTU/kg diets increased most of the nutrients' digestibility in the short-term but only P digestibility in the long-term trial. Tibia mineralization increased linearly with phytase addition (p < 0.05) only in the long-term trial. In conclusion, the effects of dietary mineral and phytase levels on growth performance are more noticeable in young animals. In addition, the duration of the trial is key due to a possible adaptation phenomenon of birds to low P supplementary levels.

Evaluation of Phosphorus Digestibility from Monocalcium and Dicalcium Phosphate Sources and Comparison between Total Tract and Prececal Digestibility Standard Methods in Broilers.

The objective of this study was to compare the total tract (total excreta and marker) and prececal methodologies to determine phosphorus (P) digestibility and to evaluate its variation as a function of the physicochemical characteristics of the inorganic phosphate used (monocalcium, MCP and dicalcium, DCP) from different commercial sources. A total of 176 1-day-old male broilers were used in two digestibility experiments. In Experiment 1, one MCP and one DCP were incorporated in the basal diet at two levels. In Experiment 2, MCP and DCP from three commercial sources were incorporated to the basal diet at one level. Physicochemical characteristics of inorganic phosphates were examined, as well. Additionally, bone mineralization and growth performance traits were investigated in both trials. The digestibility of MCP ranged from 75.2 to 87.4% and from 80.5 to 86.6% for DCP amongst methodologies, but differences between total tract and preceal methodologies were not statistically significant. Particle size, surface area, degree of crystallinity and impurities varied amongst commercial sources. The P digestibility of the three tested commercial sources of MCP was 79.6% (MCP1), 70.2% (MCP2) and 65.6% (MCP3); p > 0.05. The P digestibility of the 3 tested commercial sources of DCP was 80.1% (DCP1), 77.4% (DCP2) and 71.4% (DCP3); p > 0.05.

Effect of Dietary Mineral Content and Phytase Dose on Nutrient Utilization, Performance, Egg Traits and Bone Mineralization in Laying Hens from 22 to 31 Weeks of Age.

A total of 192 laying hens were used to evaluate the effect of dietary mineral content and phytase dose on nutrient utilization, egg production and quality and bone mineralization of young laying hens. Four dietary treatments were studied: PC, positive control with no added phytase, 4.07% Ca and 0.61% P; NC, negative control with no added phytase, 2.97% Ca and 0.37% P; and P500 and P1000, where NC diet was supplemented with phytase at 500 and 1000 FTU/kg, respectively. Hens' performance and egg traits were controlled from 22 to 31 weeks of age. Coefficients of total tract apparent digestibility (CTTAD) of nutrients were determined at 25 and 31 weeks of age. Apparent ileal digestibility (AID) and blood content of Ca and P, as well as bone traits, were determined at 31 weeks of age. Ca and P retention was higher in birds on PC diet at 25 weeks, but not at 31 weeks of age compared to those on NC diet (p < 0.05). P1000 birds had the highest CTTAD values for dry and organic matter at both ages (p < 0.001). CTTAD of Ca was significantly higher in P1000 diet than in NC diet at 31 weeks of age (p < 0.001). Birds fed with P500 diet at 25 weeks of age and P1000 at 31 weeks of age showed higher CTTAD and retention of P, but lower excretion of P than those fed NC diet (p < 0.05). Phytase inclusion linearly increased AID of dry matter and P (p < 0.001). P500 hens fed had the greatest body weight at the end of the trial (p < 0.05) and P1000 birds had the best feed conversion ratio (p < 0.05). Fowl fed a PC diet produced eggs with higher shell thickness and yolk color than those fed on NC diet (p < 0.05). Phytase inclusion linearly increased the yolk color (p < 0.05). Tibia of laying hens fed with PC had significantly higher ash content than those on NC diet (p < 0.05), and birds fed with P1000 presented intermediate values. It can be concluded that it would be advisable to increase the dose of phytase in the feed of laying hens to obtain long-term benefits.

A proposal of a new algorithm for decision-making approaches in open reduction and internal fixation of complex tibial plateau fractures - SOTA algorithm (Spanish Orthopaedic Trauma Association).

INTRODUCTION: The correct choice of surgical approaches in complex tibial plateau fractures is essential to achieve adequate reduction and fixation. Detailed fracture morphology and direction of columns displacement, that we have named the Main Deformity Direction (MDD), may aid in selecting the optimal surgical strategy. In this article we present a new algorithm based on MDD and column concepts. The aim of this study was to evaluate the outcomes of a group of tibial plateau fractures treated according to this algorithm. The secondary aim was to evaluate the incidence of the different MDD in our multicolumnar subgroup. PATIENTS AND METHODS: Excluding isolated lateral one-column fractures, open fractures and patients not treated following this algorithm, 72 patients with tibial plateau fractures surgically treated with open reduction and internal fixation were collected retrospectively, from three trauma centers, from January 2015 to April 2019. Radiological assessment was performed to establish the columns involved and the MDD. Quality and maintenance of reduction and alignment, complications and functional outcomes were assessed. RESULTS: Initial fracture analysis was performed in 72 patients (8 one-column, 35 two-column and 29 multicolumnar fractures). In the multicolumnar group, the posteromedial MDD was the most frequent pattern (17 of 29 patients). Four patients were excluded due to loss of follow-up, resulting 68 patients for final outcome analysis (7 one-column, 34 two-column and 27 multicolumnar). The average follow-up was 18 months (range: 6-52). Excellent/good outcomes were obtained in all one-column, 31 of 34 two-column and 25 of 27 multicolumnar fractures. Incomplete reduction was present in three patients. As complications, we had two loss of reduction, one conversion to knee arthroplasty, one nonunion and one deep infection. No patient presented neurological or vascular problems, knee instability or extension deficit. CONCLUSION: Satisfactory results have been obtained following the principles of our algorithm. In addition to the anatomical involvement of columns and segments, we believe that identifying the Main Deformity Direction (MDD) provides useful information for decision-making, especially in multicolumnar fractures. The most frequent MDD in our multicolumnar subgroup was the posteromedial MDD, but more than one-third presented a different MDD, requiring different surgical strategies.

Effects of Orange Pulp Conservation Methods (Dehydrated or Ensiled Sun-Dried) on the Nutritional Value for Finishing Pigs and Implications on Potential Gaseous Emissions from Slurry.

The inclusion of orange pulp (OP) in pig diets may promote the circular economy, but drying procedures might influence its nutritional value and environmental impact. The purpose of this study was to determine the energy value and nutrient digestibility of dehydrated (DOP) and ensiled sun dried (ESDOP) orange pulp. The potential ammonia (NH3) and methane (CH4) emissions derived from slurry were also measured. Digestible energies of 14.2 and 13.2 MJ/kg DM for DOP and ESDOP, respectively, were estimated by difference after a 500 g/kg substitution of a basal diet with OPs. A high fiber digestion efficiency was observed for both OPs. Pigs fed the basal diet showed a higher intake and a greater excretion of urine N than pigs fed with OP, but fecal N excretion did not differ among diets. A higher benzoic and hippuric acid content in urine was observed in OP than in basal diet. Altogether, these findings explained a lower pH in slurry of OP diets and a reduction of potential NH3 emissions. The biochemical CH4 potential also decreased, especially with ESDOP. Overall, both OP are relevant sources of energy for pig diets. Their inclusion in feeds generate favorable changes of slurry characteristics that reduce potential NH3 and CH4 emissions.

Effects of Exercise Sequence and Velocity Loss Threshold During Resistance Training on Following Endurance and Strength Performance During Concurrent Training.

PURPOSE: This study aimed to analyze the response to 4 concurrent training interventions differing in the training sequence and in the velocity loss (VL) threshold during strength training (20% vs 40%) on following endurance and strength performance. METHODS: A randomized crossover research design was used. Sixteen trained men performed 4 training interventions consisting of endurance training (ET) followed by resistance training (RT), with 20% and 40% VL, respectively (ET + RT20 and ET + RT40), and RT with 20% and 40% VL, respectively, followed by ET (RT20 + ET and RT40 + ET). The ET consisted of running for 10 minutes at 90% of maximal aerobic velocity. The RT consisted of 3 squat sets with 60% of 1-repetition maximum. A 5-minute rest was given between exercises. The oxygen uptake throughout the ET and repetition velocity during RT were recorded. The blood lactate concentration, vertical jump, and squat velocity were measured at preexercise and after the endurance and strength exercises. RESULTS: The RT40 + ET protocol showed an impaired running time along with higher ventilatory equivalents compared with those protocols that performed the ET without previous fatigue. No significant differences were observed in the repetitions per set performed for a given VL threshold, regardless of the exercise sequence. The protocols consisting of 40%VL induced greater reductions in jump height and squat velocity, along with elevated blood lactate concentration. CONCLUSIONS: A high VL magnitude (40%VL) induced higher metabolic and mechanical stress, as well as greater residual fatigue, on the following ET performance.

HIV-1 resistance patterns to integrase inhibitors in Chilean patients with virological failure on raltegravir-containing regimens.

In this viewpoint we would like to describe our results in terms of resistance pattern in Chilean patients with virological failure (VF) on raltegravir (RAL)-containing-regimens and highlight the need for the concomitant availability of genotypic resistance testing to integrase strand transfer inhibitors (INSTIs) introduction in antiretroviral regimens, particularly in countries in South America. Indeed we found in our study the presence of two or more primary mutations in some of the participants which is associated with cross-resistance to all INSTIs. By using timely genotyping, we could optimally manage these patients, early after detection of VF.

Standardization of a colorimetric technique for determination of enzymatic activity of diamine oxidase (DAO) and its application in patients with clinical diagnosis of histamine intolerance.

BACKGROUND: Diamine Oxidase (DAO) has an essential role for degradation of exogenous histamine in the intestine; thus, histamine intolerance (HI) mainly has been correlated to a low concentration and/or activity of this enzyme. The objective of the study was to standardize a colorimetric technique to measure the enzymatic activity (function) of hDAO to then apply it to a series of 22 patients with a clinical diagnosis of HI. METHODS: For the standardization variables such as volume and type of sample, incubation time, wavelength of maximum absorption, types of substrates, and concentration of oxidized ascorbate were evaluated. Then the activity and concentration of DAO was determined in 22 patients diagnosed with HI and 22 healthy subjects. RESULTS: The mean of serum DAO concentration in the 22 patients was of 9.268 ± 1.124 U/mL. The mean of serum DAO concentration in the 22 controls was of 20.710 ± 2.509 U/mL, being significantly higher (P value 0.0002) the mean of the samples. The mean of serum DAO activity of the patients was of 1.143 ± 0.085 U/L and the controls was 1.533 ± 0.119 U/L, significantly greater than the patients (P value 0.011). In addition, the sensitivity of both techniques was 0.63. In the measuring of DAO concentration the specificity was 0.9, constituting a good diagnostic test, especially to rule out the true negatives. The determination of DAO activity had a specificity of 0.68. CONCLUSIONS: Although we used a small number of patients and controls and the absorbance values were lower than expected, statistically significant differences were found in the levels of concentration and DAO activity between the patients with histamine intolerance and the controls. Therefore, the measuring of DAO concentration and DAO activity is a good diagnostic strategy for study suspect cases of HI. The simultaneous use of both assays allows to reduce positive and negative false results, for example, patients with normal DAO levels that could present a dysfunction in the activity of this enzyme.

Adherence to antihypertensive drug treatment in Argentina: A multicenter study.

Adherence to antihypertensive medication is an important challenge that doctors often face in the treatment of hypertension. Good adherence is crucial to prevent cardiovascular complications. In consequence, the present study aimed at determining the prevalence of adherence to antihypertensive treatment and identifying associated clinical variables. A multicenter cross-sectional study was conducted in 12 cities of Argentina. A systematic sampling was performed in order to select patients with hypertension and under pharmacological treatment for at least 6 months. Physicians took three BP measurements, and the level of adherence was assessed using the self-administered Morisky questionnaire (MMAS-8). Participants were classified into three levels of adherence: high adherence-MMAS score of 8; medium adherence-MMAS scores of 6 to <8; and low adherence-MMAS scores of <6. A total of 1111 individuals (62 ± 12 years old, women 49.4%) were included in the present analysis; 159 (14.3%), 329 (29.6%) and 623 (56.1%) patients had low, medium, and high adherence, respectively. The prevalence of controlled hypertension increased only in high adherent patients: 42.8%, 42.2%, and 64.5% for low, medium, and high adherence groups, respectively. Similarly, systolic BP was lower only in the high adherence group. High educational level (OR 3.47, 95% CI 2.68-4.49) and diuretic treatment (OR 0.64, 95% CI 0.47-0.88) were independent predictors of high adherence. In conclusion, more than a half of treated hypertensive patients had a high level of adherence. These patients had lower BP values and higher control levels. A high educational level predicts high adherence.

Nanostructured porous graphene and its composites for energy storage applications.

Graphene, 2D atomic-layer of sp2 carbon, has attracted a great deal of interest for use in solar cells, LEDs, electronic skin, touchscreens, energy storage devices, and microelectronics. This is due to excellent properties of graphene, such as a high theoretical surface area, electrical conductivity, and mechanical strength. The fundamental structure of graphene is also manipulatable, allowing for the formation of an even more extraordinary material, porous graphene. Porous graphene structures can be categorized as microporous, mesoporous, or macroporous depending on the pore size, all with their own unique advantages. These characteristics of graphene, which are further explained in this paper, may be the key to greatly improving a wide range of applications in energy storage systems.

Functional outcome after calcaneus fractures: a comparison between polytrauma patients and isolated fractures.

BACKGROUND: Displaced intra-articular calcaneal fractures (DIACFs) can have long-term consequences in terms of foot pain and disability. This injury frequently occurs in the context of polytrauma patients and its influence in long-term functional prognosis is well known. The purpose of this study is to compare the etiology, severity, and functional outcome of the operated DIACFs between polytrauma patients and isolated cases. METHODS: Eighty-six patients with operated displaced intra-articular calcaneal fractures through open reduction and internal fixation (ORIF) were managed at our institution between January 1, 2008 and December 31, 2015. Eighty patients completed the follow-up, nine of them with both calcaneus operated (89 calcaneus) by ORIF with a plate. Two groups were established; one included 11 (14%) polytrauma patients, three of them with operated bilateral calcaneus, and 69 (86%) patients without polytrauma diagnosis. Severity of trauma, injury patterns, psychiatric background, associated fractures, second surgeries and functional questionnaire (adjusted American Orthopedic Foot and Ankle Society - AOFAS - ankle-hindfoot scale and SF-36) were collected. A detailed comparative statistical analysis is provided. RESULTS: There was significant statistical relationship between polytrauma patients and those with psychiatric comorbidities, severe trauma or severe injury, but not with second surgeries, later subtalar arthrodesis or outcome measures. Only calcaneal fractures with comminution according to Sanders classification was associated with second surgeries and later subtalar arthrodesis. The AOFAS score was solely related to trauma severity (71.5 vs 77; p = 0.29) and calcaneal fracture's comminution according to Sanders classification (79.4 type II and 79.3 type III vs 69.2 type IV; p = 0.000 and p = 0.008, respectively) and SF-36 score only with trauma severity (67.4 vs 78.1; p = 0.00) and psychiatric comorbidities (62.2 vs 75.8; p = 0.048). CONCLUSION: No differences in outcome measures and second surgeries were found between polytrauma patients and isolated fractures. For DIACFs, the severity of the impact was correlated with a higher number of second surgeries and worse functional outcomes. Following a DIACF, patients with psychiatric comorbidities presented worse health-related quality of life than people without this background.

The use of agricultural substrates to improve methane yield in anaerobic co-digestion with pig slurry: effect of substrate type and inclusion level.

Anaerobic co-digestion of pig slurry with four agricultural substrates (tomato, pepper, persimmon and peach) was investigated. Each agricultural substrate was tested in co-digestion with pig slurry at four inclusion levels: 0%, 15%, 30% and 50%. Inclusion levels consisted in the replacement of the volatile solids (VS) from the pig slurry with the VS from the agricultural substrate. The effect of substrate type and inclusion level on the biochemical methane potential (BMP) was evaluated in a batch assay performed at 35 °C for 100 days. Agricultural substrate's chemical composition was also analyzed and related with BMP. Additionally, Bacteria and Archaea domains together with the four main methanogenic archaeal orders were quantified using quantitative real-time TaqMan polymerase chain reaction (qPCR) at the end of the experiment to determine the influence of agricultural substrate on sludge's microbial composition. Results showed that vegetable substrates (pepper and tomato) had higher lipid and protein content and lower carbohydrates than fruit substrates (persimmon and peach). Among substrates, vegetable substrates showed higher BMP than fruit substrates. Higher BMP values were obtained with increasing addition of agricultural substrate. The replacement of 50% of VS from pig slurry by tomato and pepper increased BMP in 41% and 44%, respectively compared with pig slurry only. Lower increments in BMP were achieved with lower inclusion levels. Results from qPCR showed that total bacteria and total archaea gene concentrations were similar in all combinations tested. Methanomicrobiales gene concentrations dominated over the rest of individual archaeal orders.

HIV-1 tropism: a comparison between RNA and proviral DNA in routine clinical samples from Chilean patients.

BACKGROUND: HIV in Chile has a notification rate of 0.01%. Coreceptor antagonists are a family of antiretroviral drugs that are used with the prior knowledge of patients HIV-1 tropism. Viral RNA-based tropism detection requires a plasma viral load ≥1000 copies/mL, while proviral DNA-based detection can be performed regardless of plasma viral load. This test is useful in patients with low or undetectable viral loads and would benefit with a proper therapy. The aim of this study was to determine the correlation between HIV RNA and proviral genotypic DNA tropism tests. FINDINGS: Forty three Chilean patients were examined using population-based V3 sequencing, and a geno2pheno false-positive rate (FPR) cutoff values of 5, 5.75, 10 and 20%. With cutoff 5.75% a concordance of 88.4% in tropism prediction was found after a simultaneous comparison between HIV tropism assessment by RNA and DNA. In total, five discrepancies (11.6%) were found, 3 patients were RNA-R5/DNA-X4 and two were RNA-X4/DNA-R5. Proviral DNA enabled the prediction of tropism in patients with a low or undetectable viral load. For cutoff 5 and 5.75% genotypic testing using proviral DNA showed a similar sensitivity for X4 as RNA. We found that the highest sensitivity for detecting the X4 strain occurred with proviral DNA and cutoff of 10 and 20%. Viral loads were higher among X4 strain carriers than among R5 strain carriers (p < 0.05). CONCLUSIONS: A high degree of concordance was found between tropism testing with RNA and testing with proviral DNA. Our results suggest that proviral DNA-based genotypic tropism testing is a useful option for patients with low or undetectable viral load who require a different therapy.

Andes virus antigens are shed in urine of patients with acute hantavirus cardiopulmonary syndrome.

Hantavirus cardiopulmonary syndrome (HCPS) is a highly pathogenic emerging disease (40% case fatality rate) caused by New World hantaviruses. Hantavirus infections are transmitted to humans mainly by inhalation of virus-contaminated aerosol particles of rodent excreta and secretions. At present, there are no antiviral drugs or immunotherapeutic agents available for the treatment of hantaviral infection, and the survival rates for infected patients hinge largely on early virus recognition and hospital admission and aggressive pulmonary and hemodynamic support. In this study, we show that Andes virus (ANDV) interacts with human apolipoprotein H (ApoH) and that ApoH-coated magnetic beads or ApoH-coated enzyme-linked immunosorbent assay plates can be used to capture and concentrate the virus from complex biological mixtures, such as serum and urine, allowing it to be detected by both immunological and molecular approaches. In addition, we report that ANDV-antigens and infectious virus are shed in urine of HCPS patients.

Immunoresponse of Coho salmon immunized with a gene expression library from Piscirickettsia salmonis.

We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 micrograms were injected intramuscularly into 60 fish followed by a second dose of 10 micrograms applied 40 days later. As control, fish were injected with the same amount of DNA of the vector pCMV-Bios without insert. The titer of IgM anti-P. salmonis of vaccinated fish, evaluated 60 days post-injection, was significantly higher than that of the control group injected with the vector alone. Moreover, this response was specific against P. salmonis antigens, since no cross reaction was detected with Renibacterium salmoninarum and Yersinia ruckeri. The vaccinated and control fish were challenged 60 days after the second dose of DNA with 2.5 x 10(7) P. salmonis corresponding to 7.5 times the LD50. At 30 days post-challenge, 100% mortality was obtained with the control fish while 20% of the vaccinated animals survived. All surviving fish exhibited a lower bacterial load in the kidney than control fish. The expression library was also tested in Balb/c mice and it was found that the humoral immune response was specific to P. salmonis and it was dependent on the amount of DNA injected.

Dengue-1 virus isolation during first dengue fever outbreak on Easter Island, Chile.

Dengue virus was detected for the first time in Chile, in an outbreak of dengue fever on Easter Island. The virus was isolated in tissue culture and characterized by reverse transcription-polymerase chain reaction as being dengue type 1.