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Introduction Advanced chronic liver disease (ACLD) is a major cause of death for people with HIV (PWH). While viral hepatitis coinfections are largely responsible for this trend, metabolic dysfunction-associated steatotic liver disease (MASLD) is an emerging concern for PWH. We aimed to assess the contribution of MASLD to incident ACLD in PWH.Methods and analysis This multicentre prospective observational cohort study will enrol 968 consecutive HIV monoinfected patients from four Canadian sites, excluding subjects with alcohol abuse, liver disease other than MASLD, or ACLD at baseline. Participants will be followed annually for 4 years by clinical evaluation, questionnaires, laboratory testing and Fibroscan to measure liver stiffness measurement (LSM) and controlled attenuation parameter (CAP). The primary outcome will be incidence of ACLD, defined as LSM>10 kPa, by MASLD status, defined as CAP≥285 dB/m with at least one metabolic abnormality, and to develop a score to classify PWH according to their risk of ACLD. Secondary outcomes will include health-related quality of life (HRQoL) and healthcare resource usage. Kaplan-Meier survival method and Cox proportional hazards regression will calculate the incidence and predictors of ACLD, respectively. Propensity score methods and marginal structural models will account for time-varying exposures. We will split the cohort into a training set (to develop the risk score) and a validation set (for validation of the score). HRQoL scores and healthcare resource usage will be compared by MASLD status using generalised linear mixed effects model.Ethics and dissemination This protocol has been approved by the ethics committees of all participating institutions. Written informed consent will be obtained from all study participants. The results of this study will be shared through scientific publications and public presentations to advocate for the inclusion of PWH in clinical trials of MASLD-targeted therapies and case-finding of ACLD in PWH.

In Chile, edible herbs are mainly grown by small farmers. This type of horticultural crop typically requires intensive management because it is highly susceptible to insects, some of which transmit viruses that severely affect crop yield and quality. In 2019, in coriander plants tested negative for all previously reported viruses, RNA-Seq analysis of one symptomatic plant revealed a plethora of viruses, including one virus known to infect coriander, five viruses never reported in coriander, and a new cytorhabdovirus with a 14,180 nucleotide RNA genome for which the species name Cytorhabdovirus coriandrum was proposed. Since all the detected viruses were aphid-borne, aphids and weeds commonly growing around the coriander field were screened for viruses. The results showed the occurrence of the same seven viruses and the alfalfa mosaic virus, another aphid-borne virus, in aphids and weeds. Together, our findings document the presence of multiple viruses in coriander and the potential role of weeds as virus reservoirs for aphid acquisition.

La investigación se centra en las líneas avanzadas desarrolladas a través de cruces interespecíficos entre Oryza sativa L. ssp. japonica x Oryza rufipogon G., y la línea tipo japónica, cuya tolerancia a las sales bajo condiciones de invernadero y campo se sometieron a evaluación. En la primera fase en invernadero, se sembraron 24 líneas avanzadas de arroz F5, cinco líneas parentales y un cultivar comercial SFL-011 (control) en medio líquido con una concentración de 7,0 dS/m CE salinizado con NaCl y control con 0,2.dS/m CE de agua de riego. En la segunda fase en condiciones de campo se evaluaron líneas de arroz F6 en suelos salinos con 7.44 dS/m CE. Se midieron variables agronómicas y de rendimiento. En condiciones de invernadero se destacaron seis líneas: Puyón/JP003 P11-106716, Puyón/JP002 P8-30552, Puyón/JP003 P11-103115, Puyón/JP002 P8-294930, JP002/JP001 P × P 5P 1322 y JP001/JP003 P1 × 11P 413, con mayor tolerancia que el control en cuanto a vigor de planta, panículas por planta, granos por panícula, peso de 1000 granos y rendimiento (g/planta). En condiciones de campo, dos líneas, Puyón/JP003 P11-106716 y Puyón/JP003 P11-103115, se destacaron por tener mejores resultados agronómicos en cultivares y rendimiento. Los resultados mostraron la importancia del uso de cruces interespecíficos entre O. Sativa ssp. japonica x O. rufipogon, y la línea tipo japónica para mejorar la tolerancia a la salinidad del arroz y garantizar altos rendimientos potenciales en suelos salinizados; de ahí la importancia del cruce en términos de rendimientos agrícolas.

Background. The growing demand for organic quinoa in the national and international market has contributed to the intensification of quinoa cultivation under conventional production systems, causing changes in economic, environmental and social terms in the quinoa-producing areas of the inter-Andean valleys of the highlands of Peru. Objective. The aim of the study was to analyze the economic, social and environmental sustainability of organic quinoa production in plots of five communities in the province of Andahuaylas, Peru. Methodology. Surveys with structured questions were applied to a stratified sample of 50 organic quinoa producers with a focus on sustainability. The internal consistency of the scales used was validated by Multiple Correspondence Analysis (MCA) using the optimal scale technique and the percentage scale was determined with 0 - 24% very low level of sustainability and 100% as optimal level of sustainability. Implications. The value of economic, environmental and social indicators determines the degree of sustainability of organic quinoa production systems. Conclusions. An Economic Indicator (KI) of 42.6%, Social Indicator (IS) of 52.7% and Environmental Indicator (EI) of 53.4% were found. The General Sustainability Index (GSI) was 50.9%, which results in an average sustainability level and with critical points related to the market and adverse climatic and biotic factors.

Background. As a consequence of the growing international quinoa demand, in the last 10 years the cultivated area in Peru has increased, from diversified production systems for self-consumption to monocultures with improved varieties compliant with market characteristics demanded. Objective. Evaluate farmer type and characteristics and their quinoa production units (QPU) based on land tenure, cultivation area, sowing intention, yield and family income. Methodology. Surveys were applied in five communities with the highest quinoa production with structured questions that incorporate economic, environmental and social variables. For the analysis of qualitative and quantitative data, statistical tools of multivariate analysis were used. To express the variables in a set of linear combinations of factors not correlated with each other, principal components analysis (PCA) was used; meanwhile, a cluster analysis was performed from the grouping of the QPU with minimum and maximum intra and inter-class variability. Implications. Knowledge of agroecosystems characteristics and the type of farmer who manages production units is the basis of all research, planning and the technology transfer processes. Conclusions. Three types of quinoa farmers and belong to family farming systems in the process of from articulation to market, were identified. The first group with limited access to land, water, and capital resources makes their exploitation difficult. Meanwhile, the second and third groups, denominated smallholders and medium holders, have land with better resources, but their difficulties persist surpluses that allow them to be capitalized. In the three groups, families carry out activities inside and outside the QPU, to generate additional income beyond their main crop.

En los últimos diez años, la producción de quinua en el Perú se ha intensificado, pasando de las prácticas tradicionales a sistemas de producción tecnificada en función de la demanda del mercado, lo que ha ocasionado cambios en términos económicos, sociales y, principalmente, ambientales. El objetivo de la investigación fue determinar los índices de sostenibilidad ambiental en tres comunidades productoras de quinua de la zona altoandina del departamento de Ayacucho, Perú. Bajo el enfoque metodológico de análisis multicriterio, se construyó una escala en porcentaje de valoración de sostenibilidad, siendo el valor cero (0) no sostenible, y 100 el umbral máximo de sostenibilidad. Para ello, se aplicaron encuestas con preguntas estructuradas en la dimensión ambiental. Los sistemas de producción de quinua alcanzan un Indicador Ambiental (IA) de 49,43 % o un rango de sostenibilidad baja. La variable de mayor contribución fue el riesgo de erosión (51,28 %), seguido del manejo de biodiversidad (48,68 %) y la conservación de la vida del suelo (48,32 %).

Premise Cañahua is a semi‐domesticated crop grown in high‐altitude regions of the Andes. It is an A‐genome diploid (2n = 2x = 18) relative of the allotetraploid (AABB) Chenopodium quinoa and shares many of its nutritional benefits. Cañahua seed contains a complete protein, a low glycemic index, and offers a wide variety of nutritionally important vitamins and minerals. Methods The reference assembly was developed using a combination of short‐ and long‐read sequencing techniques, including multiple rounds of Hi‐C–based proximity‐guided assembly. Results The final assembly of the ~363‐Mbp genome consists of 4633 scaffolds, with 96.6% of the assembly contained in nine scaffolds representing the nine haploid chromosomes of the species. Repetitive element analysis classified 52.3% of the assembly as repetitive, with the most common repeat identified as long terminal repeat retrotransposons. MAKER annotation of the final assembly yielded 22,832 putative gene models. Discussion When compared with quinoa, strong patterns of synteny support the hypothesis that cañahua is a close A‐genome diploid relative, and thus potentially a simplified model diploid species for genetic analysis and improvement of quinoa. Resequencing and phylogenetic analysis of a diversity panel of cañahua accessions suggests that coordinated efforts are needed to enhance genetic diversity conservation within ex situ germplasm collections.

The aim of the current study was to evaluate the effect of feeding Pelibuey sheep on diet supplemented with different doses of organic selenium (Se)-enriched yeast on carcasses microbiological contamination and meat physical characteristics. The experiment was conducted during the finishing stage of 18 female sheep and lasted for 60 days. In a complete randomized design, sheep were distributed to one of three treatments: the control without Se-yeast (T1), the control supplemented with Se-yeast at 0.35 mg Se/kg DM (T2), and control supplemented with Se-yeast at 0.60 mg Se/kg DM (T3). The yeast product used was Selyeast 3000TM yeast (LFA Lesaffre, Toluca, Mexico) with a Se concentration of 3000 ppm (mg/kg). Lambs were slaughtered at the end of the experiment at an average weight of 39.5±4.41 kg and samples were taken for microbiological analysis. There were no differences between treatments (P>0.05) and the aerobic plate counts for T1, T2 and T3 had indexes of 0.10, 0.08 and 0.08 log10 CFU/cm2, respectively. Total coliform counts obtained were 0.13, 0.10 and 0.09 log10 CFU/cm2 for T1, T2 and T3, respectively, and the faecal coliform counts were 0.09 log10 CFU/cm2 for T1, 0.06 log10 CFU/cm2 for T2 and 0.07 log10 CFU/cm2 for T3. No significant effects (P>0.05) were observed for carcasses physical characteristics of microbial growth, initial and ultimate pH and temperature, colour values and water holding capacity. It can therefore be concluded that organic Se-enriched yeast did not affect carcasses bacterial proliferation or meat physical characteristics.

OBJETIVO: Descreve-se a auditoria médica realizada entre as mulheres que se inscreveram em programa de pré-natal com o objetivo de verificar as características da assistência à gestação e de estabelecer diretrizes de atenção. MÉTODOS: Realizou-se levantamento epidemiológico dos cartões das gestantes inscritas no Programa de Pré-natal do Posto de Saúde da Vila Municipal, Pelotas, RS, com data provável de parto durante 1997 e no primeiro semestre de 1998. Foram incluídas as mulheres ingressadas no Programa até o quarto mês de gestação e que tiveram cinco consultas no mínimo. Foi utilizada a análise bivariada para detectar as condições marcadoras da assistência médica. RESULTADOS: Em 1997, 73 mulheres se inscreveram no Programa de Pré-natal e, em 1998, 75. A média de consultas durante o pré-natal foi de 5,2 em 1997 e 6,2 em 1998. Essa diferença foi significativa (p

Este estudo, mediante auditoria médica, avalia a qualidade da atenção médica no Posto de Saúde da Vila Municipal, Pelotas, Rio Grande do Sul. Por meio do levantamento de dados registrados nos prontuários de família em 1992, 1993 e 1994, foram coletadas informações referentes às atividades do Programa de Atenção Integral à Saúde da Mulher (PAISM), como, por exemplo, a idade da paciente, número de consultas médicas e não médicas, aferição de pressão arterial, se as mamas haviam sido examinadas, cobertura do exame citopatológico e uso de método contraceptivo. Verificou-se que, em média, as mulheres consultam-se neste serviço 3,5 a quatro vezes por ano. Destas, 53% encontram-se na faixa etária entre vinte e 39 anos de idade. Quanto à cobertura de exame citopatológico, encontrou-se registro de 42,5% procedimentos realizados nos últimos três anos. Entre as mulheres que tiveram suas mamas examinadas, 19,4% foram registradas. A aferição da tensão arterial foi o procedimento estudado que atingiu a cobertura mais elevada nesta auditoria. Acredita-se que o cuidado ao paciente também depende da qualidade do registro; portanto, a baixa qualidade dos registros médicos encontrados servirá para fomentar discussões com a equipe, a fim de contribuir para um melhor atendimento médico, beneficiando os pacientes.

Background: Patient engagement with transmission-based precautions can be an important strategy to prevent adverse events related to isolation. Most patient education is still highly prescriptive and is thus unlikely to help. Effective communication requires behavior change, leading to a meaningful dialog between the parties involved., Objective: evaluate implementation process of a protocol for effective communication with patients in transmission-based precautions (Com-Efe)., Methods: Implementation research using qualitative methods in 4 sequential phases: (1) nonparticipant observation in inpatient wards; (2) design of the intervention for implementation; (3) adaptation of the Com-Efe through workshops with nurses; (4) final assessment of the implementation results through interviews with nurses. Study was performed in a public, secondary, teaching hospital. Consolidated Framework for Implementation Research was used as the reference for interview design and data analysis, aiming to identify barriers and enablers of the implementation process., Results: Main factors that could have facilitated adherence were beliefs and perceived advantages in using the Com-Efe by nurses. Main barriers that may have contributed to the failure were the unfavorable climate for implementation, insufficient individual and leadership commitment, and the lack of understanding of the concepts underpinning effective communication., Conclusions: Despite using a systematic approach, the Com-Efe protocol was not fully implemented. The lessons learned in this study allowed us to propose suggestions for future protocol implementations in similar contexts., (Copyright © 2022. Published by Elsevier Inc.)

Purpose: To describe the implementation pro cess and the preliminary results of a surveillance system for healthcare-associated endophthalmitis., Methods: This is a case study of the implementation of a surveillance system for healthcare-associated endophthalmitis. The system for healthcare-associated endophthalmitis is a structured system that enables surveillance of cases of healthcare-associated endophthalmitis after intraocular procedures, developed and coordinated by the Division of Hospital Infection at the State Health Department, São Paulo, Brazil. The implementation process included a pilot phase, followed by a scaling-up phase. Data were reported monthly to the Division of Hospital Infection by participating healthcare facilities that performed intraocular procedures in the state of São Paulo, Brazil, from September 2017 to December 2019., Results: Among the 1,483 eligible healthcare facilities, 175 engaged in the study (participation rate of 11.8%), reporting 222,728 intraocular procedures performed, of which 164,207 were cataract surgery and 58,521 were intravitreal injections. The overall incidence rate of endophthalmitis was reported to be 0.05% (n=105; 80 cases after cataract surgery and 25 cases after intravitreal injections). The incidence rates for healthcare facilities ranged from 0.02% to 4.55%. Most cases were caused by gram-positive bacteria, mainly Staphylococcus spp. In 36 (46.2%) of the cases, there was no bacterial growth; no sample was collected in 28 (26.7%) cases. This system for healthcare-associated endophthalmitis enabled the identification of an outbreak of four cases of endophthalmitis after intravitreal injections., Conclusion: The system for healthcare-associated endophthalmitis proved to be operationally viable and efficient for monitoring cases of endophthalmitis at the state level.

Background: In healthcare implementation research, there is little discussion of researchers' experiences of using frameworks, including the Consolidated Framework for Implementation Research (CFIR)., Aim: To identify and discuss the benefits and challenges encountered and the lessons learnt from researchers' experiences of using the CFIR in different contexts and phases of research., Discussion: This article synthesises the reflections of nursing and public health researchers on their experiences of using the CFIR across four separate healthcare-associated infection prevention and control implementation studies. The CFIR's benefits and challenges, the resolutions to these challenges and the lessons learnt from the application of the framework were discussed., Conclusions: Identified benefits included the framework's adaptability and flexibility, and its provision of structure and shared language for research. Translation to another language and differentiating between domains and constructs were challenges., Implications for Practice: Nurse researchers may find this article useful when considering use of the CFIR, or to anticipate and prepare to overcome the challenges highlighted when using the framework., Competing Interests: The authors declare that there is no conflict of interest. The authors received no financial support for the authorship or publication of this article, (©2022 RCN Publishing Company Ltd. All rights reserved. Not to be copied, transmitted or recorded in any way, in whole or part, without prior permission of the publishers.)

Objective: This study evaluated religiosity and its association with mental health, quality of life, and the intensity of pelvic pain in women with chronic pelvic pain., Methods: A cross-sectional study was conducted with 100 women with chronic pelvic pain. Religiosity was investigated using the Duke University Religion Index. Quality of life was evaluated using the abbreviated version of the World Health Organization's quality of life instrument. Depression and anxiety were assessed using the Hospital Anxiety and Depression Scale, while pain intensity was evaluated using a visual analog scale., Results: Almost half the participants attended religious services at least once a week and 62% prayed, meditated, or studied the Bible at least once a day. There was no association between religiosity and anxiety or depression. The intrinsic religiosity score was lower for women with mixed anxiety-depressive disorder compared to those without mixed anxiety-depressive disorder. There was a positive association between intrinsic religiosity and the psychological health domain of the quality of life instrument. There was no association between religiosity and pain intensity., Conclusions: Women with chronic pelvic pain were strongly religious. Women with mixed anxiety-depressive disorder had lower levels of intrinsic religiosity. On the other hand, intrinsic religiosity was positively associated with quality of life in women with chronic pelvic pain. Religiosity was not associated with the intensity of pelvic pain. These data suggest that health-care professionals should take religiosity into account when treating women with chronic pelvic pain.

The use of nanoparticulate systems with action in breast and ovarian cancer has been highlighted in recent years as an alternative to increasing the therapeutic index of conventional anticancer drugs. Thus, nanoparticles have advantageous characteristics in the treatment of cancer. Several nanocarriers of drugs and nanoparticles are described in the literature. The pharmacokinetics of the drugs can be modified by the use of nanocarriers, which in turn facilitate the specific delivery of the drug to the tumour cell. Therefore, the present work is a review that examines some nanosystems with nanoparticles for action in the treatment of breast cancer and ovarian cancer.

An analytical model to describe diffusion of oligonucleotides from stable hydrogel beads is developed and experimentally verified. The synthesized alginate beads are Fe(3+) -cross-linked and polyelectrolyte-doped for uniformity and stability at physiological pH. Data on diffusion of oligonucleotides from inside the beads provide physical insights into the volume nature of the immobilization of a fraction of oligonucleotides due to polyelectrolyte cross-linking, that is, the absence of a surface-layer barrier in this case. Furthermore, the results suggest a new simple approach to measuring the diffusion coefficient of mobile oligonucleotide molecules inside hydrogels. The considered alginate beads provide a model for a well-defined component in drug-release systems and for the oligonucleotide-release transduction steps in drug-delivering and biocomputing applications. This is illustrated by destabilizing the beads with citrate, which induces full oligonucleotide release with nondiffusional kinetics., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The diagnosis of malaria, caused by Plasmodium spp., still remains a challenging process. Especially in low-income countries, a rapid user-friendly method is needed for the efficient care of the patient. A small-angle light scattering device consisting of hardware and software was developed. Using the DNA-binding dye SYBR Green, malaria infections could be distinguished in healthy red blood cells infected with Plasmodium. Subsequently, samples from parasite positive and negative patients living in a hyper-endemic area of Kinshasa, DRC were assessed. The scatter profiles were distinct and malaria infection could be detected using the Giemsa stain. Although these results are preliminary, they indicate that the device has the potential to be used as a new diagnostic tool for the detection of Malaria infection., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Intermittent preventive treatment (IPT) is a proven malaria control strategy in infants and pregnancy. School-aged children represent 26 % of the African population, and an increasing percentage of them are scholarized. Malaria is causing 50 % of deaths in this age group and malaria control efforts may shift the malaria burden to older age groups. Schools have been suggested as a platform for health interventions delivery (deworming, iron-folic acid, nutrients supplementation, (boost-)immunization) and as a possible delivery system for IPT in schoolchildren (IPTsc). However, the current evidence on the efficacy and safety of IPTsc is limited and the optimal therapeutic regimen remains controversial., Methods: A systematic search for studies reporting efficacy and safety of IPT in schoolchildren was conducted using PubMed, Web of Science, Clinicaltrials and WHO/ICTRP database, and abstracts from congresses with the following key words: intermittent, preventive treatment AND malaria OR Plasmodium falciparum AND schoolchildren NOT infant NOT pregnancy., Results: Five studies were identified. Most IPTsc regimes demonstrated substantial protection against malaria parasitaemia, with dihydroartemisinin-piperaquine (DP) given monthly having the highest protective effect (PE) (94 %; 95 % CI 93-96). Contrarily, SP did not provide any PE against parasitaemia. However, no IPT regimen provided a PE above 50 % in regard to anaemia, and highest protection was provided by SP+ amodiaquine (AQ) given four-monthly (50 %; 95 % CI 41-53). The best protection against clinical malaria was observed in children monthly treated with DP (97 %; 95 % CI 87-98). However, there was no protection when the drug was given three-monthly. No severe adverse events were associated with the drugs used for IPTsc., Conclusion: IPTsc may reduce the malaria-related burden in schoolchildren. However, more studies assessing efficacy of IPT in particular against malaria-related anaemia and clinical malaria in schoolchildren must be conducted.

Supramolecular self-assembly has been demonstrated to be a useful approach to developing new functional nanomaterials. In this work, we used a cobalt Prussian blue analogue (PBA, Co3[Co(CN)6]2) compound and a β-cyclodextrin (CD) macrocycle to develop a novel host-guest PBA-CD nanomaterial. The preparation of the functional magnetic material involved the self-assembly of CD molecules onto a PBA surface by a co-precipitation method. According to transmission electronic microscopy results, PBA-CD exhibited a polydisperse structure composed of 3D nanocubes with a mean edge length of 85 nm, which became shorter after CD incorporation. The supramolecular arrangement and structural, crystalline and thermal properties of the hybrid material were studied in detail by vibrational and electronic spectroscopies and X-ray diffraction. The cyclic voltammogram of the hybrid material in a 0.1 mol · L(-1) NaCl supporting electrolyte exhibited a quasi-reversible redox process, attributed to Co2+/Co3+ conversion, with an E1/2 value of 0.46 V (vs. SCE), with higher reversibility observed for the system in the presence of CD. The standard rate constants for PBA and PBA-CD were determined to be 0.07 and 0.13 s(-1), respectively, which suggests that the interaction between the nanocubes and CD at the supramolecular level improves electron transfer. We expect that the properties observed for the hybrid material make it a potential candidate for (bio)sensing designs with a desirable capability for drug delivery.

Direct electron transfer (DET) between redox enzymes and electrode surfaces is of growing interest and an important strategy in the development of biofuel cells and biosensors. Among the nanomaterials utilized at electrode/enzyme interfaces to enhance the electronic communication, graphene oxide (GO) has been identified as a highly promising candidate. It is postulated that GO layers decrease the distance between the flavin cofactor (FAD/FADH2) of the glucose oxidase enzyme (GOx) and the electrode surface, though experimental evidence concerning the distance dependence of the rate constant for heterogeneous electron-transfer (k(het)) has not yet been observed. In this work, we report the experimentally observed DET of the GOx enzyme adsorbed on flexible carbon fiber (FCF) electrodes modified with GO (FCF-GO), where the k(het) between GO and electroactive GOx has been measured at a structurally well-defined interface. The curves obtained from the Marcus theory were used to obtain k(het), by using the model proposed by Chidsey. In agreement with experimental data, this model proved to be useful to systematically probe the dependence of electron transfer rates on distance, in order to provide an empirical basis to understand the origin of interfacial DET between GO and GOx. We also demonstrate that the presence of GO at the enzyme/electrode interface diminishes the activation energy by decreasing the distance between the electrode surface and FAD/FADH2.

Background: In areas of high malaria transmission, Plasmodium falciparum infection during pregnancy is characterized by malaria-related anaemia, placental malaria and does not always result in clinical symptoms. This situation is associated with poor pregnancy outcomes. The aim of this study was to determine the extent of asymptomatic P. falciparum infection, its relation with anaemia as well as the most cost-effective technique for its diagnosis in healthy pregnant women living in Kinshasa, Democratic Republic of the Congo., Methods: In a cross-sectional study design, information on socio-demographic characteristics and cost data were collected in healthy pregnant women attending antenatal care consultations. Plasmodium falciparum infection was diagnosed using rapid diagnostic test (RDT), microscopy and polymerase chain reaction (PCR). Haemoglobin concentration was also determined., Results: In total, 332 pregnant women were enrolled. RDT and microscopy data were available for all the blood samples and 166 samples were analysed by PCR. The prevalence of asymptomatic P. falciparum infection using microscopy, RDTs and PCR, were respectively 21.6%, 27.4% and 29.5%. Taking PCR as a reference, RDTs had a sensitivity of 81.6% and a specificity of 94.9% to diagnose asymptomatic P. falciparum infection. The corresponding values for microscopy were 67.3% and 97.4%. The prevalence of anaemia was 61.1% and asymptomatic malaria increased five times the odds (p < 0.001) of having anaemia. RDTs were more cost-effective compared to microscopy. Incremental cost-effectiveness ratio was US$ 63.47 per microscopy adequately diagnosed case., Conclusion: These alarming results emphasize the need to actively diagnose and treat asymptomatic malaria infection during all antenatal care visits. Moreover, in DRC, malaria and anaemia control efforts should be strengthened by promoting the use of insecticide-treated nets, intermittent preventive treatment with sulphadoxine-pyrimethamine and iron and folic acid supplements.

Purpose: To investigate the presence of depressive symptoms in women with chronic pelvic pain., Methods: This descriptive cross-sectional study was performed with women aged 18 years or older, diagnosed with chronic pelvic pain, with no pregnancy history in the previous year, and with no cancer history. The sample was established by calculating the representative sample, estimated as 50 women. All women were undergoing treatment at a gynecology outpatient clinic, referred by the primary health care network of the Brazilian national health system. Data collection was performed from October 2009 to May 2010. The women's sociodemographic, economic and clinical characteristics were analyzed. Pain intensity was evaluated using a visual analogue scale. The depressive symptoms were investigated using Beck's Depression Inventory. Statistical analysis was performed using position measures (mean, median), dispersion (standard deviation) and the χ² test. Values of p ≤ .05 were considered statistically significant., Results: The participants' mean age was 41.6 ± 9.4 years. The following features predominated: secondary education level; pardo (brown) skin color; Catholic religion; and living with a steady partner. Most (98%) were economically active and worked with general domestic services. Regarding the participants' subjective perception of pain, 52% reported experiencing intense pain, while 48% reported experiencing moderate pain. Most women (52%) had been living with pain for five years or less, and 30%, for over 11 years. The mean BDI score was 17.4 (± 9.4). It was observed that 58% of the women presented mild, moderate and severe depressive symptoms according to the BDI. The most frequent depressive symptoms were fatigability, loss of libido, irritability, difficulty to work, somatic preoccupations, crying, dissatisfaction, sadness, and insomnia., Conclusion: Depressive symptoms were frequent among these women suffering with chronic pelvic pain.

Aims: Stress mechanisms paradoxically contribute to allergic episodes in humans and mice. Glucocorticoids (GC) and interleukin (IL)-5 synergically upregulate murine bone-marrow eosinophil production. Here we explored the role of endogenous GC in allergen-stimulated bone-marrow eosinophil production in ovalbumin-sensitized/challenged mice., Main Methods: In BALB/c or C57BL/6 mice, sensitized and intranasally challenged with ovalbumin, we monitored eosinophil numbers in freshly harvested or cultured bone-marrow, and plasma corticosterone levels. Metyrapone (MET) was used to inhibit GC synthesis, and RU486 to block GC actions. In sensitized mice challenged intraperitoneally, we examined the relationship between eosinophilia of bone-marrow and peritoneal cavity, in the absence or presence of RU486. In experiments involving in vivo neutralization of tumor necrosis factor-α (TNF) by specific antibodies, or using mice which lack functional type I TNF receptors (TNFRI), we evaluated the relationship between TNF blockade, corticosterone levels, RU486 or MET treatment and challenge-induced bone-marrow eosinophilia., Key Findings: RU486 or MET pretreatments abolished challenge-induced increases in eosinophil numbers in bone-marrow (in vivo and ex vivo), and in the peritoneal cavity. MET, but not RU486, prevented the challenge-induced increase in corticosterone levels. Challenge-induced bone-marrow eosinophilia and corticosterone surge were abolished in TNFRI-deficient mice. Anti-TNF-treatment very effectively prevented challenge-induced bone-marrow eosinophilia, in the absence of RU486 or MET, but had no independent effect in the presence of either drug., Significance: Endogenous GC was essential for allergen challenge-induced increases in eosinophil numbers inside bone-marrow. This effect required TNF and TNFRI, which suggests an immunoendocrine mechanism., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The roles of eosinophils in antimicrobial defense remain incompletely understood. In ovalbumin-sensitized mice, eosinophils are selectively recruited to the peritoneal cavity by antigen, eotaxin, or leukotriene(LT)B4, a 5-lipoxygenase (5-LO) metabolite. 5-LO blockade prevents responses to both antigen and eotaxin. We examined responses to eotaxin in the absence of sensitization and their dependence on 5-LO. BALB/c or PAS mice and their mutants (5-LO-deficient ALOX; eosinophil-deficient GATA-1) were injected i.p. with eotaxin, eosinophils, or both, and leukocyte accumulation was quantified up to 24 h. Significant recruitment of eosinophils by eotaxin in BALB/c, up to 24 h, was accompanied by much larger numbers of recruited neutrophils and monocytes/macrophages. These effects were abolished by eotaxin neutralization and 5-LO-activating protein inhibitor MK886. In ALOX (but not PAS) mice, eotaxin recruitment was abolished for eosinophils and halved for neutrophils. In GATA-1 mutants, eotaxin recruited neither neutrophils nor macrophages. Transfer of eosinophils cultured from bone-marrow of BALB/c donors, or from ALOX donors, into GATA-1 mutant recipients, i.p., restored eotaxin recruitment of neutrophils and showed that the critical step dependent on 5-LO is the initial recruitment of eosinophils by eotaxin, not the secondary neutrophil accumulation. Eosinophil-dependent recruitment of neutrophils in naive BALB/c mice was associated with increased binding of bacteria.

In Democratic Republic of Congo access to health care is limited because of many geographical and financial barriers, while quality of care is often low. Global health donors assist the country with a number of community-oriented interventions such as free distribution of bednets, antihelminthic drugs, vitamin A supplementation and vaccination campaigns, but uptake of these interventions is not always optimal. The aim of this study was to explore the perceptions of poor urban communities of the capital Kinshasa with regard to health issues in general as well as their experiences and expectations concerning facility-based health services and community-oriented health interventions. Applying an approach rooted in the grounded theory framework, focus group discussions were conducted in eight neighborhoods of poor urban areas in the city of Kinshasa in July 2011. Study participants were easily able to evoke the city's major health problems, with the notable exceptions of malnutrition and HIV/AIDS. They perceive the high out-of-pocket cost of health services as the major obstacle when seeking access to quality care. Knowledge of ongoing community-oriented health interventions seems good. Still, while the study participants agree that those interventions are beneficial; their acceptability seems to be problematic. This is chiefly put down to a lack of information and government communication about the programs and their interventions. Furthermore, the study participants referred to rumors and the deterring effect of stories about alleged harmful consequences of those interventions. Along with improving the provision and quality of general health care, the government and international actors must improve their efforts in informing the communities about disease control programs, their rationale and benefit/risk ratio. Directly engaging community members in a dialogue might be beneficial in terms of improving acceptability and overall access to health services and interventions. Novel ways of reducing the high out-of-pocket expenditure should also be explored.

This article describes an investigation to identify risk factors for postoperative endophthalmitis caused by Pseudomonas aeruginosa. A retrospective case-control study, including 8 cases and 24 controls operated from 2005 to 2009, was undertaken. The risk factors associated with P aeruginosa infection were a particular operating room (P = .003) and longer phacoemulsification duration (P = .001), suggesting that infection was not associated with patient characteristics but rather with the surgical environment and phacoemulsification duration., (Copyright © 2013 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)

Up- and downregulation of eosinopoiesis control pulmonary eosinophilia in human asthma. In mice, eosinopoiesis is suppressed in vitro by prostaglandin E2 (PGE2) and in vivo by diethylcarbamazine, through a proapoptotic mechanism sequentially requiring inducible NO synthase (iNOS) and the ligand for death receptor CD95 (CD95L). We examined the roles of iNOS, cAMP-mediated signaling, caspases, and CD95L/CD95 in suppression of eosinopoiesis by PGE2 and other agents signaling through cAMP. Bone-marrow collected from BALB/c mice, or from iNOS-, CD95-, or CD95L-deficient mutants (and wild-type controls), was cultured with interleukin-5 (IL-5), alone or associated with PGE2, cAMP-inducing/mimetic agents, caspase inhibitor zVAD-fmk, iNOS inhibitor aminoguanidine, or combinations thereof, and eosinopoiesis was evaluated at various times. PGE2, added up to 24 hours of culture, dose-dependently suppressed eosinopoiesis, by inducing apoptosis. This effect was (a) paralleled by induction of iNOS in eosinophils; (b) duplicated by sodium nitroprusside, isoproterenol, and cAMP-inducing/mimetic agents; (c) prevented by protein kinase A inhibition. NO was produced through iNOS by dibutyryl-cAMP-stimulated bone-marrow. Overall, PGE2 and isoproterenol shared a requirement for four effector elements (iNOS, CD95L, CD95, and terminal caspases), which together define a pathway targeted by several soluble up- and downmodulators of eosinopoiesis, including drugs, mediators of inflammation, and cytokines.

Purpose: To evaluate the cytotoxicity of reusable cannulas for ophthalmic surgery after the cannulas were filled with an ophthalmic viscosurgical device (OVD) and cleaned with an enzymatic detergent., Setting: Microbiological Testing Laboratory, Department of Medical-Surgical Nursing, University of São Paulo School of Nursing, and Cell Culture Section, Adolfo Lutz Institute, São Paulo, Brazil., Design: Experimental study., Methods: The sample consisted of 30 reusable 25-gauge injection cannulas, 20.0 mm in length, whose lumens were filled with an OVD solution for 50 minutes. The following steps were used to process the cannulas: (1) presoaking, (2) washing the lumen using a high-pressure water jet, (3) backwashing with enzymatic detergent in ultrasonic cleaner, (4) preliminary rinsing with tap water, (5) final rinsing with sterile distilled water, (6) drying with compressed filtered air, (7) wrapping in surgical-grade paper, and (8) steam sterilization at 134°C for 4 minutes. The cannulas were then tested for cytotoxicity according to the United States Pharmacopeia 32., Results: The cleaning protocol used in this study removed residues of OVD solution and enzymatic detergent as shown by the lack of cytotoxicity of all sample extracts., Conclusion: This cleaning protocol has the potential to minimize the occurrence of toxic anterior segment syndrome associated with residues of OVD solutions and enzymatic detergents., Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned., (Copyright © 2013 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.)

Context and Objective: Neonatal sepsis is associated with premature birth and maternal infection. Large-scale studies seek to define markers that identify neonates at risk of developing sepsis. Here, we examine whether the scientific evidence supports systematic use of polymorphism genotyping in cytokine and innate immunity genes, to identify neonates at increased risk of sepsis., Design and Setting: Narrative literature review conducted at Fernandes Figueira Institute, Brazil., Methods: The literature was searched in PubMed, Embase (Excerpta Medica Database), Lilacs (Literatura Latino-Americana e do Caribe em Ciências da Saúde), SciELO (Scientific Electronic Library Online) and Cochrane Library. From > 400,000 references, 548 were retrieved based on inclusion/exclusion criteria; 22 were selected for detailed analysis after quality assessment., Results: The studies retrieved addressed the impact of gene polymorphisms relating to immune mechanisms (most often TNF-a, LT-a, IL-6, IL-1β, IL-1ra, L-selectin, CD14 and MBL) or inflammatory mechanisms (ACE and angiotensin II receptors; secretory PLA2; and hemostatic factors). Despite initial reports suggesting positive associations between specific polymorphisms and increased risk of sepsis, the accumulated evidence has not confirmed that any of them have predictive power to justify systematic genotyping., Conclusions: Sepsis prediction through systematic genotyping needs to be reevaluated, based on studies that demonstrate the functional impact of gene polymorphisms and epidemiological differences among ethnically distinct populations.

Allergic airway inflammation is attenuated by oral tolerization (oral exposure to allergen, followed by conventional sensitization and challenge with homologous antigen), which decreases airway allergen challenge-induced eosinophilic infiltration of the lungs and bone marrow eosinophilia. We examined its effects on bone marrow eosinophil and neutrophil production. Mice of wild type (BP-2, BALB/c, and C57BL/6) and mutant strains (lacking iNOS or CD95L) were given ovalbumin (OVA) or water (vehicle) orally and subsequently sensitized and challenged with OVA (OVA/OVA/OVA and H2O/OVA/OVA groups, resp.). Anti-OVA IgG and IgE, bone marrow eosinophil and neutrophil numbers, and eosinophil and neutrophil production ex vivo were evaluated. T lymphocytes from OVA/OVA/OVA or control H2O/OVA/OVA donors were transferred into naïve syngeneic recipients, which were subsequently sensitized/challenged with OVA. Alternatively, T lymphocytes were cocultured with bone marrow eosinophil precursors from histocompatible sensitized/challenged mice. OVA/OVA/OVA mice of the BP-2 and BALB/c strains showed, relative to H2O/OVA/OVA controls, significantly decreased bone marrow eosinophil counts and ex vivo eosinopoiesis/neutropoiesis. Full effectiveness in vivo required sequential oral/subcutaneous/intranasal exposures to the same allergen. Transfer of splenic T lymphocytes from OVA/OVA/OVA donors to naive recipients prevented bone marrow eosinophilia and eosinopoiesis in response to recipient sensitization/challenge and supressed eosinopoiesis upon coculture with syngeneic bone marrow precursors from sensitized/challenged donors.

The aim of this study was to evaluate the microbial growth on single-use vitrectomy probes reprocessed in healthcare practice. We investigated nine vitrectomy probes that had been reused and reprocessed using different methods. The samples were sectioned, individually, in portions of 3.5 cm, totaling 979 sampling units (extensions, connectors and vitrectomy cutters), which were inoculated in culture medium and incubated at 37º C for 14 days. The results showed microbial growth on 57 (5.8%) sample units, 25 of which had been sterilized using ethylene oxide, 16 by hydrogen peroxide plasma, and 16 by low-temperature steam and formaldehyde. Seventeen microbial species were identified. The most prevalent were: Micrococcus spp., coagulase-negative Staphylococcus, Pseudomonas spp., and Bacillus subtilis. The reuse of single-use vitrectomy probes was shown to be unsafe, therefore this practice is not recommended.

Accumulating evidence indicates that genetic background influences the outcome of sepsis, which despite medical advances continues to be a major cause of morbidity and mortality. This study aimed to evaluate the influence of SNPs LTA +252A>G, TNF-863C>A and TNF-308G>A on susceptibility to sepsis, acute respiratory distress syndrome (ARDS), septic shock and sepsis mortality. A prospective case-control study was carried out in a Brazilian pediatric intensive care unit and included 490 septic pediatric patients submitted to mechanical ventilation and 610 healthy children. No SNP association was found with respect to sepsis susceptibility. Nevertheless, a haplotype was identified that was protective against sepsis (+252A/-863A/-308G; OR=0.65; p=0.03). We further observed protection against ARDS in TNF-308 GA genotype carriers (OR=0.29; p=0.0006) and -308A allele carriers (OR=0.40; p=0.003). In addition, increased risk for ARDS was detectable with the TNF-863 CA genotype (OR=1.83; p=0.01) and the -863A carrier status (OR=1.82; p=0.01). After stratification according to age, this outcome remained significantly associated with the -308GA genotype in infants. Finally, protection against sepsis-associated mortality was found for the TNF-308 GA genotype (OR=0.22; p=0.04). Overall, our findings document a protective effect of the TNF-308 GA genotype for the ARDS and sepsis mortality outcomes, further providing evidence for an increased risk of ARDS associated with the TNF-863 CA genotype. Trial registration (www.clinicaltrials.gov): NCT00792883., (Copyright © 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

This article describes a postoperative endophthalmitis (POE) surveillance system in place in a specialized ophthalmologic center in São Paulo, Brazil. The study involved a review of medical records from 2004-2009, during which a total of 31,999 intraocular surgeries were performed. Nineteen of these cases fulfilled the criteria for POE, for an infection rate of 0.06%. The main etiologic agent causing POE was Pseudomonas aeruginosa, identified in 42.1% of the cases (8/19)., (Copyright © 2012 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)

Paromomycin (PMM) has recently been introduced for treatment of visceral leishmaniasis in India. Although no clinical resistance has yet been reported, proactive vigilance should be warranted. The present in vitro study compared the outcome and stability of experimental PMM-resistance induction on promastigotes and intracellular amastigotes. Cloned antimony-resistant L. donovani field isolates from India and Nepal were exposed to stepwise increasing concentrations of PMM (up to 500 µM), either as promastigotes or intracellular amastigotes. One resulting resistant strain was cloned and checked for stability of resistance by drug-free in vitro passage as promastigotes for 20 weeks or a single in vivo passage in the golden hamster. Resistance selection in promastigotes took about 25 weeks to reach the maximal 97 µM inclusion level that did not affect normal growth. Comparison of the IC(50) values between the parent and the selected strains revealed a 9 to 11-fold resistance for the Indian and 3 to 5-fold for the Nepalese strains whereby the resistant phenotype was also maintained at the level of the amastigote. Applying PMM pressure to intracellular amastigotes produced resistance after just two selection cycles (IC(50) = 199 µM) compared to the parent strain (IC(50) = 45 µM). In the amastigote-induced strains/clones, lower PMM susceptibilities were seen only in amastigotes and not at all in promastigotes. This resistance phenotype remained stable after serial in vitro passage as promastigote for 20 weeks and after a single in vivo passage in the hamster. This study clearly demonstrates that a different PMM-resistance phenotype is obtained whether drug selection is applied to promastigotes or intracellular amastigotes. These findings may have important relevance to resistance mechanism investigations and the likelihood of resistance development and detection in the field.

Leishmaniasis is a neglected tropical disease that affects about 350 million individuals worldwide. The protozoan parasite has a relatively simple life cycle with two principal stages: the flagellated mobile promastigote living in the gut of the sandfly vector and the intracellular amastigote within phagolysosomal vesicles of the vertebrate host macrophage. This review presents a state-of-the-art overview of the redox biology at the parasite-macrophage interface. Although Leishmania species are susceptible in vitro to exogenous superoxide radical, hydrogen peroxide, nitric oxide, and peroxynitrite, they manage to survive the endogenous oxidative burst during phagocytosis and the subsequent elevated nitric oxide production in the macrophage. The parasite adopts various defense mechanisms to cope with oxidative stress: the lipophosphoglycan membrane decreases superoxide radical production by inhibiting NADPH oxidase assembly and the parasite also protects itself by expressing antioxidant enzymes and proteins. Some of these enzymes could be considered potential drug targets because they are not expressed in mammals. In respect to antileishmanial therapy, the effects of current drugs on parasite-macrophage redox biology and its involvement in the development of drug resistance and treatment failure are presented., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Aims: Granulocyte Colony-Stimulating Factor (G-CSF), which mobilizes hemopoietic stem cells (HSC), is believed to protect HSC graft recipients from graft-versus-host disease by enhancing Th2 cytokine secretion. Accordingly, G-CSF should aggravate Th2-dependent allergic pulmonary inflammation and the associated eosinophilia. We evaluated the effects of G-CSF in a model of allergic pulmonary inflammation., Main Methods: Allergic pulmonary inflammation was induced by repeated aerosol allergen challenge in ovalbumin-sensitized C57BL/6J mice. The effects of allergen challenge and of G-CSF pretreatment were evaluated by monitoring: a) eosinophilia and cytokine/chemokine content of bronchoalveolar lavage fluid, pulmonary interstitium, and blood; b) changes in airway resistance; and c) changes in bone-marrow eosinophil production., Key Findings: Contrary to expectations, G-CSF pretreatment neither induced nor enhanced allergic pulmonary inflammation. Instead, G-CSF: a) suppressed accumulation of infiltrating eosinophils in bronchoalveolar, peribronchial and perivascular spaces of challenged lungs; and b) prevented ovalbumin challenge-induced rises in airway resistance. G-CSF had multiple regulatory effects on cytokine and chemokine production: in bronchoalveolar lavage fluid, levels of IL-1 and IL-12 (p40), eotaxin and MIP-1a were decreased; in plasma, KC, a neutrophil chemoattractant, was increased, while IL-5 was decreased and eotaxin was unaffected. In bone-marrow, G-CSF: a) prevented the increase in bone-marrow eosinophil production induced by ovalbumin challenge of sensitized mice; and b) selectively stimulated neutrophil colony formation., Significance: These observations challenge the view that G-CSF deviates cytokine production towards a Th2 profile in vivo, and suggest that this neutrophil-selective hemopoietin affects eosinophilic inflammation by a combination of effects on lung cytokine production and bone-marrow hemopoiesis., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objectives: The oleanane triterpene saponin PX-6518, with known potent in vitro and in vivo activity against Leishmania donovani, was investigated for its spectrum against the cutaneous species Leishmania mexicana, Leishmania panamensis and Leishmania major., Methods: In vitro activity was based on the reduction of amastigotes in primary peritoneal mouse macrophages. BALB/c mice were injected with 2 × 10(6) amastigotes in the base of the tail (L. panamensis and L. major) or the foot (L. mexicana) and subcutaneously treated with PX-6518 [1-10 mg/kg body weight (BW)] or Pentostam(®) (250 mg/kg BW Sb(V) eq). Evolution of skin lesions was monitored in a prophylactic dose-finding study, and early curative [6 weeks post-infection (pi)] and late curative (>8-10 weeks pi) studies., Results: While moderate susceptibility to PX-6518 was obtained in vitro (IC(50): 1-4 µg/mL), excellent in vivo activity was demonstrated. In the prophylactic study (six administrations on alternate days, starting at 1 day pi), PX-6518 was 100% effective at 1 mg/kg BW against L. mexicana and L. panamensis, whereas L. major lesions could be prevented at 2 mg/kg BW. In the early curative (1 mg/kg BW once a week for 4 weeks) and late curative (1 mg/kg BW twice a week for 4 weeks) studies, PX-6518 completely healed L. mexicana and L. panamensis lesions, whereas L. major lesions were reduced by ∼ 50%., Conclusions: This study demonstrates that PX-6518 possesses potent and broad-spectrum prophylactic and curative efficacy against cutaneous leishmaniasis in the BALB/c mouse model. L. major was the least susceptible species tested and parasitological cure could not be obtained.

A reliable, rapid and low-cost method for drug sensitivity determination of Giardia duodenalis trophozoites (WB-strain) was developed in a 96-well plate. Using a standard inoculum of 5 x 10(4) trophozoites per well (300 microl), good growth was obtained after sealing the plate with an air-tight adhesive tape and incubation at 37 degrees C for 72 h in modified TYI-S-33 medium. Viable burdens were quantified using the formazan dyes MTT (100 microg/well) and XTT (20 microg/well) and the fluorescent substrate resazurin (2.5 microg/well). Prior removal of the culture medium is required since it causes spontaneous reduction of the substrate. Resazurin proved to be far superior to MTT and XTT with a level of sensitivity of about 3 x 10(4) trophozoites. Inhibitory concentrations (IC(50)) of several anti-giardial reference drugs were within the range of published values: metronidazole 2.25 microM, tinidazole 1.75 microM, albendazole 0.10 microM, furazolidone 2.00 microM and quinacrine 0.32 microM. The broad-spectrum antibiotics chloramphenicol, rifampicin, penicillin G+streptomycin and gentamycin were devoid of any inhibitory activity and are considered suitable for decontamination during excystation experiments.

Because low tumor necrosis factor-alpha (TNF-alpha) production has been reported in malnourished children, in contrast with high production of TNF-alpha in experimental protein-energy malnutrition, we reevaluated the production of TNF-alpha in whole blood cultures from children with primary malnutrition free from infection, and in healthy sex- and age-matched controls. Mononuclear cells in blood diluted 1:5 in endotoxin-free medium released TNF-alpha for 24 h. Spontaneously released TNF-alpha levels (mean +/- SEM), as measured by enzyme immunoassay in the supernatants of unstimulated 24-h cultures, were 10,941 +/- 2,591 pg/ml in children with malnutrition (N = 11) and 533 +/- 267 pg/ml in controls (N = 18) (P < 0.0001). TNF-alpha production was increased by stimulation with lipopolysaccharide (LPS), with maximal production of 67,341 +/- 16,580 pg/ml TNF-alpha in malnourished children and 25,198 +/- 2,493 pg/ml in controls (P = 0.002). In control subjects, LPS dose-dependently induced TNF-alpha production, with maximal responses obtained at 2000 ng/ml. In contrast, malnourished patients produced significantly more TNF-alpha with 0.02-200 ng/ml LPS, responded maximally at a 10-fold lower LPS concentration (200 ng/ml), and presented high-dose inhibition at 2000 ng/ml. TNF-alpha production a) was significantly influenced by LPS concentration in control subjects, but not in malnourished children, who responded strongly to very low LPS concentrations, and b) presented a significant, negative correlation (r = -0.703, P = 0.023) between spontaneous release and the LPS concentration that elicited maximal responses in malnourished patients. These findings indicate that malnourished children are not deficient in TNF-alpha production, and suggest that their cells are primed for increased TNF-alpha production.

Background: Platelet-derived growth factor (PDGF) is a mitogen and chemoattractant for cells of mesenchymal origin. Over-expression of PDGF-B can promote formation of inflammatory lesions in the lungs of transgenic mice. Moreover, continuous exposure to PDGF inhibits collagen production by osteoblastic cells. Thus, the expression of mitogenic factors in an inflammatory context may limit the differentiated function of cells, and thereby limit repair following periodontal attachment and bone loss. The goals of the present study were to test whether PDGF is present at increased levels in inflamed gingiva and to localize its expression in gingival biopsies from individuals with chronic periodontitis., Methods: Tissues obtained during therapeutic procedures from inflamed and control sites of 9 patients were subjected to protein extraction, descriptive histology by hematoxylin and eosin, or immunohistochemistry assays. Quantification was calculated with an enzyme-linked immunosorbent assay (ELISA) kit specific for PDGF-AB. For the immunolocalization, anti-PDGF-A and -B antibodies were employed., Results: PDGF concentration in the total protein extract was approximately 3 times higher in the inflamed sites (0.60 +/- 0.18 ng/mg versus 0.20 +/- 0.05 ng/mg; P = 0.03). Immunohistochemistry revealed prominent expression of PDGF in the pocket epithelial cells as well as the adjacent connective tissue. In contrast, little or no expression was detected in control biopsies devoid of the pocket epithelium and granulation tissue., Conclusions: PDGF is present in increased levels in the human inflamed gingiva and is mainly localized to the pocket epithelium. It is possible that chronic expression of PDGF contributes to the inflammatory changes that occur during periodontal diseases.