Development and Characterization of an Allergic Asthma Rat Model for Interventional Studies.

Allergic asthma is one of the most common chronic diseases of the airways, however it still remains underdiagnosed and hence undertreated. Therefore, an allergic asthma rat model would be useful to be applied in future therapeutic strategy studies. The aim of the present study was to develop an objective model of allergic asthma in atopic rats that allows the induction and quantification of anaphylactic shock with quantitative variables. Female Brown Norway rats were intraperitoneally sensitized with ovalbumin (OVA), alum and Bordetella pertussis toxin and boosted a week later with OVA in alum. At day 28, all rats received an intranasal challenge with OVA. Anaphylactic response was accurately assessed by changes in motor activity and body temperature. Leukotriene concentration was determined in the bronchoalveolar lavage fluid (BALF), and total and IgE anti-OVA antibodies were quantified in blood and BALF samples. The asthmatic animals' motility and body temperature were reduced after the shock for at least 20 h. The asthmatic animals developed anti-OVA IgE antibodies both in BALF and in serum. These results show an effective and relatively rapid model of allergic asthma in female Brown Norway rats that allows the quantification of the anaphylactic response.

Biological Markers to Predict Outcome in Mechanically Ventilated Patients with Severe COVID-19 Living at High Altitude.

BACKGROUND: There is not much evidence on the prognostic utility of different biological markers in patients with severe COVID-19 living at high altitude. The objective of this study was to determine the predictive value of inflammatory and hematological markers for the risk of mortality at 28 days in patients with severe COVID-19 under invasive mechanical ventilation, living at high altitude and in a low-resource setting. METHODS: We performed a retrospective observational study including patients with severe COVID-19, under mechanical ventilation and admitted to the intensive care unit (ICU) located at 2850 m above sea level, between 1 April 2020 and 1 August 2021. Inflammatory (interleukin-6 (IL-6), ferritin, D-dimer, lactate dehydrogenase (LDH)) and hematologic (mean platelet volume (MPV), neutrophil/lymphocyte ratio (NLR), MPV/platelet ratio) markers were evaluated at 24 h and in subsequent controls, and when available at 48 h and 72 h after admission to the ICU. The primary outcome was the association of inflammatory and hematological markers with the risk of mortality at 28 days. RESULTS: We analyzed 223 patients (median age (1st quartile [Q1]-3rd quartile [Q3]) 51 (26-75) years and 70.4% male). Patients with severe COVID-19 and with IL-6 values at 24 h ≥ 11, NLR values at 24 h ≥ 22, and NLR values at 72 h ≥ 14 were 8.3, 3.8, and 3.8 times more likely to die at 28 days, respectively. The SOFA and APACHE-II scores were not able to independently predict mortality. CONCLUSIONS: In mechanically ventilated patients with severe COVID-19 and living at high altitude, low-cost and immediately available blood markers such as IL-6 and NLR may predict the severity of the disease in low-resource settings.

Physicochemical and Rheological Characteristics of Commercial and Monovarietal Wheat Flours from Peru.

In Peru, wheat (Triticum aestivum L.) is one of the main resources in the food industry; however, due to its low harvested area, it is the second most imported cereal. The quality of wheat flour was studied to verify that it has desirable characteristics for the preparation of bakery products. The quality of commercial and monovarietal wheat flours was assessed by measuring their physicochemical and rheological parameters, as well as the gluten content and wheat protein fractions. Eight commercial wheat flours and four monovarietal wheat flours (Barba negra, Candeal, Espelta, and Duro) from Peru were evaluated. Commercial wheat flours presented significantly higher levels of protein and gluten index compared to monovarietal wheat flours (p < 0.05). Between both groups, no significant differences were observed in the content of wet and dry gluten. Interestingly, monovarietal wheat flours presented a higher percentage of gliadins and albumins/globulins, as well as lower levels of glutenin, compared to commercial wheat flours (p < 0.05). According to the logistic regression models, the baking strength (W) was the most important parameter to evaluate the quality of commercial and monovarietal wheat flours. Our results show that monovarietal wheat flours show a lower quality compared to commercial wheat flours.

Production of Protein Hydrolysate from Quinoa (Chenopodium quinoa Willd.): Economic and Experimental Evaluation of Two Pretreatments Using Supercritical Fluids' Extraction and Conventional Solvent Extraction.

Supercritical fluids' extraction (SFE) and conventional solvent extraction (CSE) for defatting of quinoa flour as pretreatments to produce the quinoa protein hydrolysate (QPH) were studied. The objective was to extract the oil and separate the phenolic compounds (PC) and the defatted quinoa flour for subsequent quinoa protein extraction and enzymatic hydrolysis. The oil extraction yield (OEY), total flavonoid content (TFC), and QPH yield were compared. SuperPro Designer 9.0® software was used to estimate the cost of manufacturing (COM), productivity, and net present value (NPV) on laboratory and industrial scales. SFE allows higher OEY and separation of PC. The SFE oil showed a higher OEY (99.70%), higher antioxidant activity (34.28 mg GAE/100 g), higher QPH yield (197.12%), lower COM (US$ 90.10/kg), and higher NPV (US$ 205,006,000) as compared to CSE (with 77.59%, 160.52%, US$ 109.29/kg, and US$ 28,159,000, respectively). The sensitivity analysis showed that the sale of by-products improves the economic results: at the industrial scale, no significant differences were found, and both processes are economically feasible. However, results indicate that SFE allows the recovery of an oil and QPH of better nutritional quality and a high level of purity-free organic solvents for further health and nutraceutical uses.

Influence of Consumption of Two Peruvian Cocoa Populations on Mucosal and Systemic Immune Response in an Allergic Asthma Rat Model.

Different cocoa populations have demonstrated a protective role in a rat model of allergic asthma by attenuating the immunoglobulin (Ig) E synthesis and partially protecting against anaphylactic response. The aim of this study was to ascertain the effect of diets containing two native Peruvian cocoa populations ("Amazonas Peru" or APC, and "Criollo de Montaña" or CMC) and an ordinary cocoa (OC) on the bronchial compartment and the systemic and mucosal immune system in the same rat model of allergic asthma. Among other variables, cells and IgA content in the bronchoalveolar lavage fluid (BALF) and serum anti-allergen antibody response were analyzed. The three cocoa populations prevented the increase of the serum specific IgG1 (T helper 2 isotype). The three cocoa diets decreased asthma-induced granulocyte increase in the BALF, which was mainly due to the reduction in the proportion of eosinophils. Moreover, both the OC and CMC diets were able to prevent the leukocyte infiltration caused by asthma induction in both the trachea and nasal cavity and decreased the IgA in both fecal and BALF samples. Overall, these results highlight the potential of different cocoa populations in the prevention of allergic asthma.

Encapsulation of Bioactive Compounds for Food and Agricultural Applications.

This review presents an updated scenario of findings and evolutions of encapsulation of bioactive compounds for food and agricultural applications. Many polymers have been reported as encapsulated agents, such as sodium alginate, gum Arabic, chitosan, cellulose and carboxymethylcellulose, pectin, Shellac, xanthan gum, zein, pullulan, maltodextrin, whey protein, galactomannan, modified starch, polycaprolactone, and sodium caseinate. The main encapsulation methods investigated in the study include both physical and chemical ones, such as freeze-drying, spray-drying, extrusion, coacervation, complexation, and supercritical anti-solvent drying. Consequently, in the food area, bioactive peptides, vitamins, essential oils, caffeine, plant extracts, fatty acids, flavonoids, carotenoids, and terpenes are the main compounds encapsulated. In the agricultural area, essential oils, lipids, phytotoxins, medicines, vaccines, hemoglobin, and microbial metabolites are the main compounds encapsulated. Most scientific investigations have one or more objectives, such as to improve the stability of formulated systems, increase the release time, retain and protect active properties, reduce lipid oxidation, maintain organoleptic properties, and present bioactivities even in extreme thermal, radiation, and pH conditions. Considering the increasing worldwide interest for biomolecules in modern and sustainable agriculture, encapsulation can be efficient for the formulation of biofungicides, biopesticides, bioherbicides, and biofertilizers. With this review, it is inferred that the current scenario indicates evolutions in the production methods by increasing the scales and the techno-economic feasibilities. The Technology Readiness Level (TRL) for most of the encapsulation methods is going beyond TRL 6, in which the knowledge gathered allows for having a functional prototype or a representative model of the encapsulation technologies presented in this review.

Comparison of Four Oil Extraction Methods for Sinami Fruit (Oenocarpus mapora H. Karst): Evaluating Quality, Polyphenol Content and Antioxidant Activity.

The sinami palm (Oenocarpus mapora H. Karst) is a plant from the South American Amazonia that has great potential for industrial applications in the development of functional foods, nutraceuticals and cosmeceuticals. In this manuscript, the physicochemical properties, total polyphenol content and antioxidant activity of sinami oil that was obtained using four extraction systems, namely expeller press extraction (EPE), cold press extraction (CPE), ultrasound-assisted extraction (UAE) and supercritical fluid extraction (SFE), were studied and compared. The oxidative stability (OSI) was statistically non-significant in EPE and SFE. The chromatic properties (CIELab) were influenced by the extraction methods and SFE presented high values of L* and a lower content of plant pigments. Ultrasound-assisted extraction showed a higher content of polyphenols and higher antioxidant activity. Different analyses for the evaluation of the physicochemical properties, the content of total polyphenols and antioxidant activity were used to classify sinami oil according to chemometrics using principal component analysis (PCA). For example, the sinami oil that was obtained using each extraction method was in a different part of the plot. In summary, sinami oil is an excellent resource for plant pigments. Additionally, the information that was obtained on the quality parameters in this study provided a good foundation for further studies on the characterization of major and minor compounds.

Effect of pretreatment by supercritical fluids on antioxidant activity of protein hydrolyzate from quinoa (Chenopodium quinoa Willd.).

The effect of two pretreatments on the antioxidant activity was evaluated in quinoa protein hydrolysate, using supercritical CO2 extraction and ethanol as cosolvent, this type of pretreatment was compared to a conventional petroleum ether extraction method without recovery of bioactive compounds. The extractions were carried out at a temperature of 55°C and a pressure of 23 MPa using ethanol (7-8 g quinoa/100 ml); the CO2 mass flow was 35 g/min, the extraction time was 240 min and the particle size was 500 µm, enzyme COROLASE® 7089 was applied for enzymatic hydrolysis, finally ABTS test assessed antioxidant activity. A significant effect was found on the degree of hydrolysis (23.93%) and antioxidant activity (1,181.64 µmol TE/g protein) compared to a conventional method (24.33%) and (1,448.84 µmol TE/g protein). In conclusion, our results suggest that the use of supercritical CO2 and the addition of ethanol as cosolvent are the interesting green technology, to recovery oil and separate phenolic compounds prior to enzymatic hydrolysis to avoid interference with biological activities from quinoa protein hydrolysates, and shows highest antioxidant activity to be incorporate in food products.

Evaluation of host genetic and viral factors as surrogate markers for HTLV-1-associated myelopathy/tropical spastic paraparesis in Peruvian HTLV-1-infected patients.

Human T-lymphotropic virus 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a complication that affects up to 5% of HTLV-1-infected individuals. Several host genetic and viral factors have been associated with the risk of HAM/TSP. The aim of this study was to evaluate the performance of a prognostic model for HAM/TSP developed in Japan in a Peruvian population of 71 HAM/TSP patients and 94 asymptomatic carriers (ACs). This model included age, proviral load (PVL), the presence of HLA-A*02 and HLA-Cw*08 alleles, SDF-1 +801, and TNF-alpha -863 polymorphisms, and viral subgroup. We describe frequencies for the four host genetic markers and demonstrate the presence of the HTLV-1 tax B subgroup in Peru. Using cross-validation, we show that the predictive ability of the prognostic model, as characterized by the area under the receiver-operating characteristic curve (AUC), does not differ from a model containing PVL only (both AUC = 0.74). We found some suggestive evidence of a protective effect of the HLA-A*02 allele but failed to replicate the associations with the other three genetic markers and with viral subgroup. A logistic model containing PVL, age, gender, and HLA-A*02 provided the best predictive ability in the Peruvian cohort (AUC = 0.79). J. Med. Virol. 82:460-466, 2010. (c) 2010 Wiley-Liss, Inc.

Attenuating Effect of Peruvian Cocoa Populations on the Acute Asthmatic Response in Brown Norway Rats.

Cocoa contains bioactive components, which vary according to genetic and environmental factors. The present study aimed to ascertain the anti-allergic properties of native Peruvian cocoa populations ("Blanco de Piura" or BPC, "Amazonas Peru" or APC, "Criollo de Montaña" or CMC, "Chuncho" or CCC, and an ordinary cocoa or OC). To do so, after an initial in vitro approach, an in vivo study focused on the induction of an anaphylactic response associated with allergic asthma in Brown Norway rats was carried out. Based on their polyphenol content, antioxidant activity and in vitro effects, the APC and CMC were selected to be included in the in vivo study. Cocoa diets were tested in a model of allergic asthma in which anaphylactic response was assessed by changes in body temperature, motor activity and body weight. The concentration of specific immunoglobulin E (IgE), mast cell protease and leukotrienes was also quantified in serum and/or bronchoalveolar lavage fluid. CMC and OC populations exhibited a protective effect on the allergic asthma rat model as evidenced by means of a partial protection against anaphylactic response and, above all, in the synthesis of IgE and the release of mast cell protease.

Phytochemical screening and DPPH radical scavenging activity of three morphotypes of Mauritia flexuosa L.f. from Peru, and thermal stability of a milk-based beverage enriched with carotenoids from these fruits.

Mauritia flexuosa L.f. is a palm tree which presents great morphological variability (morphotypes), represented mainly by the mesocarp color of its fruits. The objective of the study was to characterize the physicochemical and antioxidant properties of three morphotypes of Mauritia flexuosa L.f. ("Yellow", "Colour" and "Shambo") of greater economic importance in the Peruvian Amazon. "Shambo" showed a significantly high content of bioactive compounds (total phenolics, flavonoids and carotenoids) and DPPH radical scavenging activity compared to the "Yellow" and "Colour" morphotypes (p ≤ 0.05). There was a significant correlation between DPPH radical scavenging activity and total phenolics, flavonoids and carotenoids (p ≤ 0.01). Furthermore, milk-based beverages enriched with carotenoids of those morphotypes of Mauritia flexuosa L.f. have been shown to be a good source of bioactive compounds for use in the food industry. The milk-based beverages enriched with carotenoids of those morphotypes of Mauritia flexuosa L.f. showed higher lightness (L∗) and yellowness (b∗).

IFN-gamma production in response to Tax 161-233, and frequency of CD4+ Foxp3+ and Lin HLA-DRhigh CD123+ cells, discriminate HAM/TSP patients from asymptomatic HTLV-1-carriers in a Peruvian population.

Human T-lymphotropic virus 1 (HTLV-1) can cause HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The objective of this study was to gain insight into the pathogenesis of HAM/TSP by focusing on the CD8(+) T-cell response. Twenty-three HTLV-1-seronegative controls (SC), 29 asymptomatic HTLV-1 carriers (AC) and 48 patients with HAM/TSP were enrolled in the study. We evaluated the production of interferon-gamma (IFN-gamma) in peripheral blood mononuclear cells stimulated with Tax overlapping peptides, the expression of genes related to the CD8(+) cytotoxic T-cell response, the frequency of CD4(+) Foxp3(+) cells and of dendritic cells, and the HTLV-1 provirus load (PVL). The frequency of cells producing IFN-gamma in response to Tax 161-233, but not to Tax 11-19, discriminated patients with HAM/TSP from AC. The increased pro-inflammatory response observed in patients with HAM/TSP was shared by AC with a high PVL, who also exhibited lower levels of granzyme H mRNA in unstimulated CD8(+) T cells than AC with a low PVL. Patients with HAM/TSP showed higher frequencies of CD4(+) Foxp3(+) cells and lower frequencies of plasmacytoid dendritic cells (pDC) than AC. Our findings are consistent with a model in which HTLV-1, along with the host genetic background, drives quantitative and qualitative changes in pDC and CD4(+) Foxp3(+) cells that lead to a predominance of inflammatory responses over lytic responses in the CD8(+) T-cell response of individuals predisposed to develop HAM/TSP.

IFN-γ Response Is Associated to Time Exposure Among Asymptomatic Immune Responders That Visited American Tegumentary Leishmaniasis Endemic Areas in Peru.

Clinical manifestations of American Tegumentary Leishmaniasis (ATL) include cutaneous (CL) and mucous forms (ML); however, there are asymptomatic individuals who despite being infected do not present any clinical manifestations. This study characterized the cell-mediated immunity of travelers who lived in the Andean highlands of Cusco, free of leishmaniasis transmission, which eventually visited leishmaniasis endemic in the Amazonian basin and returned home without any clinical signs of the disease. Their immune response was compared with CL and ML patients who acquired the disease during their stage in the same region. Fifty-four human subjects from the highlands of Cusco (Peru), who have visited an endemic area, were enrolled: 28 of them did not show any symptoms, 12 showed CL and 14 showed ML. Ten healthy subjects from a non-endemic area (HS) were included as controls. T-cell proliferation was evaluated using peripheral blood mononuclear cells (PBMC) stimulated for 5 days with a total soluble leishmanial antigen (TSLA) of L. (V.) braziliensis. Th1/Th2/Th17 cytokines were also quantified in the supernatants by a flow cytometry multiplex assay. T-cell proliferation was expressed as stimulation index (SI) and the cut off was fixed at SI >2.47. Fifteen out of 28 subjects did not show any signs of disease (54%); subjects with an SI above the cut off. They were defined as asymptomatic immune responders (AIR). CL and ML patients presented a higher SI than HS and AIR. Among the latter group, the exposure time to Leishmania was clearly associated with the IFN-γ response. Increased levels of this cytokine were observed in individuals who remained <90 days in an endemic area of leishmaniasis. Our results evidenced two sub-populations among asymptomatic individuals, one AIR who did not develop clinical disease manifestations when they were exposed to Leishmania in endemic areas. Exposure time to Leishmania in the wild was associated with the IFN-γ response.

Immunological response in cases of complicated and uncomplicated bartonellosis during pregnancy.

Bartonellosis (Carrion's Disease) during pregnancy is associated with high rates of maternal and perinatal mortality. We report the immunological patterns in two cases of human bartonellosis during pregnancy. One patient had an uncomplicated course while the second patient developed life threatening anasarca and cardiac tamponade. The patient with a complicated course had a Th1 response with a higher elevation of IL-10. This elevation has been associated with poor outcome pregnancies during bacterial infections.

Design of a predicted MHC restricted short peptide immunodiagnostic and vaccine candidate for Fowl adenovirus C in chicken infection.

Fowl adenoviruses (FAdVs) are the ethiologic agents of multiple pathologies in chicken. There are five different species of FAdVs grouped as FAdV-A, FAdV-B, FAdV-C, FAdV-D, and FAdV-E. It is of interest to develop immunodiagnostics and vaccine candidate for Peruvian FAdV-C in chicken infection using MHC restricted short peptide candidates. We sequenced the complete genome of one FAdV strain isolated from a chicken of a local farm. A total of 44 protein coding genes were identified in each genome. We sequenced twelve Cobb chicken MHC alleles from animals of different farms in the central coast of Peru, and subsequently determined three optimal human MHC-I and four optimal human MHC-II substitute alleles for MHC-peptide prediction. The potential MHC restricted short peptide epitope-like candidates were predicted using human specific (with determined suitable chicken substitutes) NetMHC MHC-peptide prediction model with web server features from all the FAdV genomes available. FAdV specific peptides with calculated binding values to known substituted chicken MHC-I and MHC-II were further filtered for diagnostics and potential vaccine epitopes. Promiscuity to the 3/4 optimal human MHC-I/II alleles and conservation among the available FAdV genomes was considered in this analysis. The localization on the surface of the protein was considered for class II predicted peptides. Thus, a set of class I and class II specific peptides from FAdV were reported in this study. Hence, a multiepitopic protein was built with these peptides, and subsequently tested to confirm the production of specific antibodies in chicken.

Genome sequence and comparative analysis of Avibacterium paragallinarum.

BACKGROUND: Avibacterium paragallinarum, the causative agent of infectious coryza, is a highly contagious respiratory acute disease of poultry, which affects commercial chickens, laying hens and broilers worldwide. METHODOLOGY: In this study, we performed the whole genome sequencing, assembly and annotation of a Peruvian isolate of A. paragallinarum. Genome was sequenced in a 454 GS FLX Titanium system. De novo assembly was performed and annotation was completed with GS De Novo Assembler 2.6 using the H. influenzae str. F3031 gene model. Manual curation of the genome was performed with Artemis. Putative function of genes was predicted with Blast2GO. Virulence factors were identified by comparison with the Virulence Factor Database. RESULTS: The genome obtained has a length of 2.47 Mb with 40.66% of GC content. Seventy five large contigs (>500 nt) were obtained, which comprised 1,204 predicted genes. All the contigs are available in Genbank [GenBank: PRJNA64665]. A total of 103 virulence factors, reported in the Virulence Factor Database, were found in A. paragallinarum. Forty four of them are present in 7 species of Haemophilus, which are related with pathogenesis, virulence and host immune system evasion. A tetracycline-resistance associated transposon (Tn10), was found in A. paragallinarum, possibly acting as a defense mechanism. DISCUSSION AND CONCLUSION: The availability of A. paragallinarum genome represents an important source of information for the development of diagnostic tests, genotyping, and novel antigens for potential vaccines against infectious coryza. Identification of virulence factors contributes to better understanding the pathogenesis, and planning efforts for prevention and control of the disease.

Cytokines and T-Lymphocute count in patients in the acute and chronic phases of Bartonella bacilliformis infection in an endemic area in peru: a pilot study.

Human Bartonellosis has an acute phase characterized by fever and hemolytic anemia, and a chronic phase with bacillary angiomatosis-like lesions. This cross-sectional pilot study evaluated the immunology patterns using pre- and post-treatment samples in patients with Human Bartonellosis. Patients between five and 60 years of age, from endemic areas in Peru, in the acute or chronic phases were included. In patients in the acute phase of Bartonellosis a state of immune peripheral tolerance should be established for persistence of the infection. Our findings were that elevation of the anti-inflammatory cytokine IL-10 and numeric abnormalities of CD4(+) and CD8(+) T-Lymphocyte counts correlated significantly with an unfavorable immune state. During the chronic phase, the elevated levels of IFN-γ and IL-4 observed in our series correlated with previous findings of endothelial invasion of B. henselae in animal models.

Short communication an interferon-γ ELISPOT assay with two cytotoxic T cell epitopes derived from HTLV-1 tax region 161-233 discriminates HTLV-1-associated myelopathy/tropical spastic paraparesis patients from asymptomatic HTLV-1 carriers in a Peruvian population.

HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a chronic and progressive disorder caused by the human T-lymphotropic virus type 1 (HTLV-1). In HTLV-1 infection, a strong cytotoxic T cell (CTL) response is mounted against the immunodominant protein Tax. Previous studies carried out by our group reported that increased IFN-γ enzyme-linked immunospot (ELISPOT) responses against the region spanning amino acids 161 to 233 of the Tax protein were associated with HAM/TSP and increased HTLV-1 proviral load (PVL). An exploratory study was conducted on 16 subjects with HAM/TSP, 13 asymptomatic carriers (AC), and 10 HTLV-1-seronegative controls (SC) to map the HAM/TSP-associated CTL epitopes within Tax region 161-233. The PVL of the infected subjects was determined and the specific CTL response was evaluated with a 6-h incubation IFN-γ ELISPOT assay using peripheral blood mononuclear cells (PBMCs) stimulated with 16 individual overlapping peptides covering the Tax region 161-233. Other proinflammatory and Th1/Th2 cytokines were also quantified in the supernatants by a flow cytometry multiplex assay. In addition, a set of human leukocyte antigen (HLA) class I alleles that bind with high affinity to the CTL epitopes of interest was determined using computational tools. Univariate analyses identified an association between ELISPOT responses to two new CTL epitopes, Tax 173-185 and Tax 181-193, and the presence of HAM/TSP as well as an increased PVL. The HLA-A*6801 allele, which is predicted to bind to the Tax 181-193 peptide, was overpresented in the HAM/TSP patients tested.

SYBR Green-based quantitation of human T-lymphotropic virus type 1 proviral load in Peruvian patients with neurological disease and asymptomatic carriers: influence of clinical status, sex, and familial relatedness.

To evaluate the human T-lymphotropic virus type 1 (HTLV-1) proviral DNA load in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and asymptomatic HTLV-1 carriers, a SYBR Green-based real-time quantitative polymerase chain reaction (qPCR) assay was developed. HTLV-1 proviral DNA in peripheral blood mononuclear cells (PBMCs) was quantified using primers targeting the pX region and the HTLV-1 copy number normalized to the amount of ERV-3 (Endogenous Retrovirus 3) cellular DNA. Thirty-three asymptomatic HTLV-1 carriers (ACs) and 39 patients with HAM/TSP were enrolled. Some participants were relatives of HAM/TSP cases (16 ACs and 7 patients with HAM/TSP). On multiple linear regression analysis, the authors found a significant association between clinical status and HTLV-1 proviral load (P < .01), but only among women. ACs showed a median proviral load of 561 copies per 104 PBMCs (interquartile range: 251-1623). In HAM/TSP patients, the median proviral load was 1783 (1385-2914). ACs related to HAM/TSP patients presented a relatively high proviral load (median 1152); however, the association between relatedness to a HAM/TSP patient and proviral load was not significant (P = .1). In HAM/TSP patients, no association was found between proviral load and disease duration, progression or severity. The fact that the effect of HAM/TSP upon the HTLV-1 proviral load differed between sexes and the finding of a high proviral load among asymptomatic relatives of HAM/TSP patients suggest that not yet identified genetic or environmental factors influence the pathogenesis of HTLV-1 infection.

Cytokines and T-Lymphocute count in patients in the acute and chronic phases of Bartonella bacilliformis infection in an endemic area in peru: a pilot study / Citoquinas y recuento de Linfocitos T en pacientes en fase aguda y crónica de infección por Bartonella bacilliformis, en una área endémica del Perú: estudio piloto

Human Bartonellosis has an acute phase characterized by fever and hemolytic anemia, and a chronic phase with bacillary angiomatosis-like lesions. This cross-sectional pilot study evaluated the immunology patterns using pre- and post-treatment samples in patients with Human Bartonellosis. Patients between five and 60 years of age, from endemic areas in Peru, in the acute or chronic phases were included. In patients in the acute phase of Bartonellosis a state of immune peripheral tolerance should be established for persistence of the infection. Our findings were that elevation of the anti-inflammatory cytokine IL-10 and numeric abnormalities of CD4+ and CD8+ T-Lymphocyte counts correlated significantly with an unfavorable immune state. During the chronic phase, the elevated levels of IFN-γ and IL-4 observed in our series correlated with previous findings of endothelial invasion of B. henselae in animal models.

La Bartonelosis Humana, tiene una fase aguda caracterizada por fiebre y anemia hemolítica, así como una fase crónica con lesiones semejantes a angiomatosis bacilar. En un estudio transversal piloto los patrones inmunológicos en pacientes con Bartonelosis Humana fueron estudiados mediante muestras pre y post tratamiento. Pacientes entre 5 y 60 años en fase aguda y crónica fueron incluidos en área endémica del Perú. En aquellos pacientes con fase aguda, una fase de tolerancia inmunológica periférica es necesaria para la persistencia de la infección. Los hallazgos de significativa elevación de citoquina anti-inflamatoria (IL-10) y anormalidades numéricas en el recuentos de Linfocitos T CD4+ y CD8+ correlacionan con un estado inmune que favorece la infección. Durante la fase crónica, elevados niveles de INF-γ y IL-4 observados en la serie de pacientes correlacionan con previos hallazgos en modelos animales que favorecen la invasión del endotelio por B. henselae.