Search

We performed a study of congenital toxoplasmosis of the first and third gestation periods in mice, and determined its effects on the embryos/fetuses, the placentae and the maternal organs. We infected pregnant BALB/c mice by i.v. injection of 2.5--10.0 × 10 6 tachyzoites of the ME49 T. gondii strain and euthanized them 72 h later. The tissues were analyzed by histopathology, immunohistochemistry and parasite-specific qPCR. Infections with the lowest dose induced remarkably different changes in the two thirds: a) all doses diminished the number of products/litter, the lowest dose only by 14%; but most embryos still visible were degenerated in the case of the first period, while the fetuses of the last third were perfectly preserved; b) the transmission rate in the first third was relatively high, but with a very low parasite burden; c) with the lowest dose, strong vascular changes (congestion, thrombosis and hemorrhage) predominated in the placentas of the first period, while they were absent in the last third; d) necrosis caused by T. gondii to maternal organs was much stronger during the last gestation period than in the first. Our results suggest that the vascular alterations at the placenta of the first third of pregnancy prevent embryo from large parasite burden, but provoke its death by starvation. In the last gestation period, there was poor control of parasite dissemination to the placenta and the fetus, but there was greater capacity of the product to defend itself from T. gondii., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: We aimed at explaining the mechanism of therapeutic effect of Umbilical Cord Mesenchymal Stem Cells (UC-MSC) in subjects with COVID-19 Acute Respiratory Distress Syndrome (ARDS). Patients with COVID-19 ARDS present with a hyperinflammatory response characterized by high levels of circulating pro-inflammatory mediators, including tumor necrosis factor α and β (TNFα and TNFβ). Inflammatory functions of these TNFs can be inhibited by soluble TNF Receptor 2 (sTNFR2). In patients with COVID-19 ARDS, UC-MSC appear to impart a robust anti-inflammatory effect, and treatment is associated with remarkable clinical improvements. We investigated the levels of TNFα, TNFβ and sTNFR2 in blood plasma samples collected from subjects with COVID-19 ARDS enrolled in our trial of UC-MSC treatment., Patients and Methods: We analyzed plasma samples from subjects with COVID-19 ARDS (n=24) enrolled in a Phase 1/2a randomized controlled trial of UC-MSC treatment. Plasma samples were obtained at Day 0 (baseline, before UC-MSC or control infusion), and Day 6 post infusion. Plasma concentrations of sTNFR2, TNFα, and TNFβ were evaluated using a quantitative multiplex protein array., Results: Our data indicate that at Day 6 after infusion, UC-MSC recipients develop significantly increased levels of plasma sTNFR2 and significantly decreased levels of TNFα and TNFβ, compared to controls., Conclusions: These observations suggest that sTNFR2 plays a mechanistic role in mediating UC-MSC effect on TNFα and TNFβ plasma levels, determining a decrease in inflammation in COVID-19 ARDS.

Objective: The aim of the present study was to compare the molecular and morphological effects of diacerein and glucosamine-chondroitin drug treatment and intra-articular injection therapy of human deciduous dental pulp stem cells (hDPSCs) in a rat knee model of induced osteoarthritis (OA)., Materials and Methods: Thirty-six adult male rats were randomly separated into six groups: Control group (without induction of OA), osteoarthritis group 60 (induction of OA, saline gavage started on day 14 and performed for 60 days, followed by euthanasia), osteoarthritis group (induction of OA and euthanasia after 14 days), diacerein group, glucosamine-chondroitin group, and mesenchymal stem cell group. The drug-treated groups were gavaged with 50 mg/kg of diacerein and 400/500 mg/kg of glucosamine-chondroitin starting on dat 14 for 60 days. The cell therapy-treated group received an intra-articular single dose of 8 × 105 hDPSCs on day 14, and euthanasia was performed after 60 days. Lateral femoral condyles were collected and prepared for immunohistochemistry and light microscopy procedures., Results: The morphological features and immunoexpression of SOX-5, IHH, MMP-8, MMP-13, and Type II collagen were statistically analysed. Our data suggest that hDPSC therapy contributes more actively and effectively in the structural reorganization of lateral femoral condyles. In contrast, the glucosamine-chondroitin sulphate treatment was more effective in inflammatory control, while diacerein showed better results associated with the maintenance of the primordial cartilage., Conclusions: The positive therapeutic effect of daily administered conventional drugs can be confirmed in a rat model of OA. However, one single dose of locally administered hDPSCs provides significant improvement in tissue regeneration in an OA model.

Inflammation represents an important factor leading to metabolic imbalance within the intervertebral disc (IVD), conducive to degenerative changes. Therefore, a thorough knowledge of the IVD and endplate (EP) cell behaviour in such pathological environments is essential when designing regenerative therapeutic strategies. The present study aimed at assessing the molecular response of the IVD constitutive nucleus pulposus (NPCs)-, annulus fibrosus (AFCs)- and endplate (EPCs)-derived cells to interleukin (IL)-1β treatment, through large-scale, high-throughput microarray and protein analysis, identifying the differentially expressed genes and released proteins. Overall, the inflammatory stimulus downregulated stemness genes while upregulating pro-inflammatory, pro-angiogenic and catabolic genes, including matrix metalloproteases, which were not balanced by a concomitant upregulation of their inhibitors. Upregulation of anti-inflammatory and anabolic tumour necrosis factor inducible gene 6 protein (TNFAIP6), of IL-1 receptor antagonist (IL-1Ra) (at gene and protein levels) and of trophic insulin-like growth factor 1 (IGF1) was also observed in all cell types; IGF1 particularly in AFCs. An overall inhibitory effect of tumour necrosis factor alpha (TNFα) signal was observed in all cell types; however, EPCs showed the strongest anti-inflammatory behaviour. AFCs and EPCs shared the ability to limit the activation of the signalling mediated by specific chemokines. AFCs showed a slightly senescent attitude, with a downregulation of genes related to DNA repair or pro-mitosis. Results allowed for the identification of specific molecular targets in IVD and EP cells that respond to an inflammatory environment. Such targets can be either silenced (when pathological targets) or stimulated to counteract the inflammation.

The coronavirus SARS-CoV-2 is cause of a global pandemic of a pneumonia-like disease termed Coronavirus Disease 2019 (COVID-19). COVID-19 presents a high mortality rate, estimated at 3.4%. More than 1 out of 4 hospitalized COVID-19 patients require admission to an Intensive Care Unit (ICU) for respiratory support, and a large proportion of these ICU-COVID-19 patients, between 17% and 46%, have died. In these patients COVID-19 infection causes an inflammatory response in the lungs that can progress to inflammation with cytokine storm, Acute Lung Injury (ALI), Acute Respiratory Distress Syndrome (ARDS), thromboembolic events, disseminated intravascular coagulation, organ failure, and death. Mesenchymal Stem Cells (MSCs) are potent immunomodulatory cells that recognize sites of injury, limit effector T cell reactions, and positively modulate regulatory cell populations. MSCs also stimulate local tissue regeneration via paracrine effects inducing angiogenic, anti-fibrotic and remodeling responses. MSCs can be derived in large number from the Umbilical Cord (UC). UC-MSCs, utilized in the allogeneic setting, have demonstrated safety and efficacy in clinical trials for a number of disease conditions including inflammatory and immune-based diseases. UC-MSCs have been shown to inhibit inflammation and fibrosis in the lungs and have been utilized to treat patients with severe COVID-19 in pilot, uncontrolled clinical trials, that reported promising results. UC-MSCs processed at our facility have been authorized by the FDA for clinical trials in patients with an Alzheimer's Disease, and in patients with Type 1 Diabetes (T1D). We hypothesize that UC-MSC will also exert beneficial therapeutic effects in COVID-19 patients with cytokine storm and ARDS. We propose an early phase controlled, randomized clinical trial in COVID-19 patients with ALI/ARDS. Subjects in the treatment group will be treated with two doses of UC-MSC (l00 × 10 6 cells). The first dose will be infused within 24 hours following study enrollment. A second dose will be administered 72 ± 6 hours after the first infusion. Subject in the control group will receive infusion of vehicle (DPBS supplemented with 1% HSA and 70 U/kg unfractionated Heparin, delivered IV) following the same timeline. Subjects will be evaluated daily during the first 6 days, then at 14, 28, 60, and 90 days following enrollment (see Schedule of Assessment for time window details). Safety will be determined by adverse events (AEs) and serious adverse events (SAEs) during the follow-up period. Efficacy will be defined by clinical outcomes, as well as a variety of pulmonary, biochemical and immunological tests. Success of the current study will provide a framework for larger controlled, randomized clinical trials and a means of accelerating a possible solution for this urgent but unmet medical need. The proposed early phase clinical trial will be performed at the University of Miami (UM), in the facilities of the Diabetes Research Institute (DRI), UHealth Intensive Care Unit (ICU) and the Clinical Translational Research Site (CTRS) at the University of Miami Miller School of Medicine and at the Jackson Memorial Hospital (JMH)., Competing Interests: Conflict of Interest All authors declare that there are no conflicts of interest regarding the publication of this manuscript.

Aims: Greenhouse gases are considered as potential atmospheric pollutants, with agriculture being one of the main emission sources. The practice of inoculating soybean seeds with Bradyrhizobium sp. might contribute to nitrous oxide (N 2 O) emissions. We analysed this capacity in five of the most used strains of Bradyrhizobium sp. in South America., Methods and Results: We analysed the denitrification pathway and N 2 O production by Bradyrhizobium japonicum E109 and CPAC15, Bradyrhizobium diazoefficiens CPAC7 and B. elkanii SEMIA 587 and SEMIA 5019, both in free-living conditions and in symbiosis with soybean. The in silico analysis indicated the absence of nosZ genes in B. japonicum and the presence of all denitrification genes in B. diazoefficiens strains, as well as the absence of nirK, norC and nosZ genes in B. elkanii. The in planta analysis confirmed N 2 O production under saprophytic conditions or symbiosis with soybean root nodules. In the case of symbiosis, up to 26.1 and 18.4 times higher in plants inoculated with SEMIA5019 and E109, respectively, than in those inoculated with USDA110., Conclusions: The strains E109, SEMIA 5019, CPAC15 and SEMIA 587 showed the highest N 2 O production both as free-living cells and in symbiotic conditions in comparison with USDA110 and CPAC7, which do have the nosZ gene. Although norC and nosZ could not be identified in silico or in vitro in SEMIA 587 and SEMIA 5019, these strains showed the capacity to produce N 2 O in our experimental conditions., Significance and Impact of the Study: This is the first report to analyse and confirm the incomplete denitrification capacity and N 2 O production in four of the five most used strains of Bradyrhizobium sp. for soybean inoculation in South America., (© 2019 The Society for Applied Microbiology.)

Diagnostic tests for toxoplasmosis are based on serological techniques due to their high sensitivity. Some IgG subclasses are related to clinical outcome in the congenital form. In this work, we determined the levels of IgG, IgA, IgG1, IgG2, IgG3 and IgG4 anti-Toxoplasma gondii antibodies in paired saliva and serum samples from 91 women by indirect ELISA using a crude extract of the RH strain. The levels of IgA, IgG2, IgG3 and IgG4 antibodies and, to a lesser extent, IgG1 did not correlate between saliva and serum, that is, most cases that were positive for one Ig class in a sample were negative or very low in the other, and vice versa. We also observed that most samples of saliva that were positive for one IgG subclass were also positive for at least 2 of the other 3; this contrasted with findings in serum, wherein each person was positive almost exclusively for one subclass, as demonstrated before by us and other researchers. Although these findings are disappointing for the use in diagnosis, the richer response in saliva might indicate local exposure to T. gondii antigens without systemic infection; thus, saliva might be reflecting a local (protective?) response against this protozoan., (© 2018 John Wiley & Sons Ltd.)

In human congenital toxoplasmosis the effects of parasite burden and pregnancy time at infection on clinical outcome are well known, but there is controversy regarding the role of Toxoplasma gondii type. Through a systematic review of the literature, we aimed to discern if T. gondii type has a role on clinical outcome in human congenital toxoplasmosis. We built up a database of congenital toxoplasmosis from reports of cases, case series and screening-based cohorts, which had information about parasite type, gestation time at maternal infection and/or clinical outcome in the product. Then, we obtained frequencies for loci used to genotype geographical origin of cases and types found. Also, odds ratios were calculated for association between time of maternal infection or parasite type on outcome. Type II parasites were the most common in Europe, Asia and Africa, while in America there were mainly atypical strains. More newborns with clinical problems were born from mothers infected during the first half of gestation than from those acquiring the parasite after week 24, regardless of parasite genotype (92.9 vs. 16.1 %, OR = 67.9, CI95 25.4-181.6). Type I and atypical parasites were associated with clinical problems as opposed to types II and III, regardless of pregnancy period at infection (86.9 vs. 72.9 %, OR = 2.47, CI95 1.1-5.4). A significant and remarkable tendency of type I parasites to be present during early pregnancy was also observed (94.4 vs. 5.6 %, P < 0.009). In addition to parasite burden and period of gestation, T. gondii genotype seems involved in CT clinical outcome.

Congenital transmission of Toxoplasma gondii may occur if the mother gets infected for the first time while pregnant. The risk of mother-to-child transmission depends on the gestation trimester at infection, being lowest in the first and highest in the last. Conversely, fetal damage is frequent and more severe at the beginning of pregnancy. The objective of this study was to evaluate congenital transmission and pathological aspects in the placenta and the fetus using a mouse model of congenital infection of the second gestation third. Forty-five female BALB/c mice were infected intravenously with 2.5-10.0 × 10(6) tachyzoites of the ME49 strain at middle gestation. Samples of maternal spleen and fetal/placental units were taken 72 h later. We determined parasite load and vertical transmission by qPCR, as well as damage macroscopically and by histopathology. With the lowest dose, 18% of the fetuses were infected. Also, 40% of fetuses/litter were altered, while this value was 10% in the control group (P < 0.05). These results are similar to those described in humans in terms of vertical transmission and fetal damage during the second third of gestation. The maternal spleen had 10-1000 times more tachyzoites than the placenta, and the later retained 90-99% of the parasites that could reach the fetus. Nevertheless, we found resorptions, abortions or fetal tissue damage in the presence but also in the absence of parasites. Our data indicate a strong protective effect of maternal organs and the placenta against fetal infection, but extensive damage of the later may led to resorption or abortion without vertical transmission., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: To test the effects of sequential exposure to FGF2, 9 and 18 on human Mesenchymal Stem Cells (hMSC) differentiation during in vitro chondrogenesis., Design: Control and FGF2-expanded hMSC were cultured in aggregates in the presence of rhFGF9, rhFGF18 or rhFGFR3-specific signaling FGF variants, starting at different times during the chondroinductive program. Quantitative real time polymerase chain reaction (qRT-PCR) and immunocytochemistry were performed at different stages. The aggregate cultures were switched to a hypertrophy-inducing medium along with rhFGFs and neutralizing antibodies against FGFR1 and FGFR3. Histological/immunohistochemical/biochemical analyses were performed., Results: FGF2-exposed hMSC during expansion up-regulated Sox9 suggesting an early activation of the chondrogenic machinery. FGF2, FGF9 and 18 modulated the expression profile of FGFR1 and FGFR3 in hMSC during expansion and chondrogenesis. In combination with transforming growth factor-beta (TGF-β), FGF9 and FGF18 inhibited chondrogenesis when added at the beginning of the program (≤ d7), while exhibiting an anabolic effect when added later (≥d14), an effect mediated by FGFR3. Finally, FGFR3 signaling induced by either FGF9 or FGF18 delayed the appearance of spontaneous and induced hypertrophy-related changes., Conclusions: The stage of hMSC-dependent chondrogenesis at which the growth factors are added impacts the progression of the differentiation program: increased cell proliferation and priming (FGF2); stimulated early chondrogenic differentiation (TGF-β, FGF9/FGF18) by shifting the chondrogenic program earlier; augmented extracellular matrix (ECM) production (FGF9/FGF18); and delayed terminal hypertrophy (FGF9/FGF18). Collectively, these factors could be used to optimize pre-implantation conditions of hMSC when used to engineer cartilage grafts., (Copyright © 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

The purpose of this study was to assess the prevalence of, and the factors associated with HIV testing and pre- and post-test counselling (PPTC) in Colombia. Cross-sectional data from the National Health Survey carried out during 2007 were analysed. Data were gathered from records of 29,760 individuals between the ages of 18 and 69 from the main regions of the country. Only 19.7% of the sample had taken an HIV test. Men, people with no education, those not affiliated with the health-care system, residents of rural areas and those aged older than 65 were less likely to have been tested for HIV; 42% of those tested did not receive pre-test counselling and 56.9% received no report of the results. Considering the low prevalence of HIV testing among the Colombian population, it is necessary to design and apply guidelines for HIV screening in all health-care settings and to conduct targeted testing in high-risk sub-populations. A national norm of PPTC in those who undergo HIV testing should be examined.

We studied the frequency of antibodies against Toxoplasma gondii in stray dogs in the city of Oaxaca, Mexico through the evaluation of 154 sera by indirect ELISA. A frequency of 61.7% was found; it was higher in males (45 of 65, 69.2%) than in females (49 of 89, 55.0%), although this difference was not statistically significant. An increase in frequency was observed with age, the lowest being among animals younger than 1 yr (4 of 20, 20.0%) and the highest in dogs older than 7 yr (21 of 25, 84.0%). This is the first study in dogs of this region of Mexico and revealed high T. gondii transmission and evidence of early exposure in animals that are in close contact with contaminated water or raw meat, or both. Further studies are needed in order to understand the role of T. gondii infection in public health.

The objective of this study was to assess the proteolytic activity of Fasciola hepatica cathepsins in liver sections from mice vaccinated with phage clones of cathepsin L mimotopes, using the film in situ zymography technique. Female BALB/c mice were immunized three times with 2.5 x 10¹¹ phage particles without adjuvant. Animals vaccinated with phage clones produced high titres of anti-mimotope antibodies and a significant reduction in fluke burden was observed following challenge with metacercariae of F. hepatica. The proteolytic activity in hepatic tissue was reduced after the immunization with phage clones.

Toxoplasma gondii infects a variety of vertebrate hosts, including humans. Transplacental passage of the parasite leads to congenital toxoplasmosis. A primary infection during the first weeks of gestation causes vertical transmission at low rate, although it causes major damage to the embryo. Transmission frequency increases to near 80% by the end of pregnancy, but the proportion of ill newborns is low. For transmission and pathogenesis, the parasite genetics is certainly important. Several host innate and adaptative immune response genes are induced during infection in adults, which control the rapidly replicating tachyzoite. The T helper 1 (Th1) response is protective, although it has to be modulated to avoid inflammatory damage. Paradoxical observations on this response pattern in congenital toxoplasmosis have been reported, as it may be protective or deleterious, inducing sterile abortion or favoring parasite transplacental passage. Regarding pregnancy, an early Th1 microenvironment is important for control of infectious diseases and successful implantation, although it has to be regulated to support trophoblast survival. Polymorphism of genes involved in these parallel phenomena, such as Toll-like receptors (TLRs), adhesins, cytokines, chemokines or their receptors, immunoglobulins or Fc receptors (FcRs), might be important in susceptibility for T. gondii vertical transmission, abortion or fetal pathology. In this study some examples are presented and discussed.

Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular obligate parasite. Its transmission has usually been attributed to ingestion of undercooked or raw meat. The frequency of T. gondii in pork, the most common meat for human consumption in Jalisco, Mexico, is unknown; in Guadalajara city high prevalence of human toxoplasmosis has been documented. Forty-eight samples of pork meat from butcher shops in Ocotlán city were analyzed. Through bioassay, 50 g of tissue was homogenized in an acidic pepsin solution and inoculated subcutaneously to previously immunosuppressed mice. Blood samples from the mice tail vein were obtained before inoculation and 7, 14, 28, and 45 days postinoculation to analyze anti-Toxoplasma immunoglobulin (Ig) M and IgG antibody kinetics by indirect enzyme-linked immunosorbent assay. For histopathology, small fragments of the brain, lungs, heart, and skeletal muscle were extracted on day 45 and were stained with hematoxylin and eosin. Also, DNA was extracted from the pork meat for PCR amplification of the B1 gene. Even though all pork samples were negative by histopathology and PCR, IgG and IgM antibodies against T. gondii were detected in 1 of the 48 inoculated mice, reflecting a frequency of 2.1% positive pork meat, which is lower than expected but similar to that found in other regions.

Objective: To detect immunoglobulin M (IgM) anti-Toxoplasma gondii antibodies and determine immunoglobulin G (IgG) titer and avidity in filter paper-embedded blood (FPEB) samples of pregnant women., Study Design: A total of 100 FPEB samples of pregnant women (30 positive and 70 negative) were analyzed for anti-T. gondii-specific IgM antibodies. Eleven and nine pairs of serum and FPEB samples were used to standardize IgG titration and avidity, respectively. Then, the correlation of avidity results was determined with 23 serum/FPEB pairs from IgG-positive cases., Result: IgM detection in FPEB was 92% sensitive and 100% specific. The titration of IgG antibodies in FPEB correlated with that of serum (r >or=0.9). Significant difference in avidity between the acute and the undetermined/chronic cases was observed in both samples. As expected, no correlation was found between IgM levels and avidity., Conclusion: The FPEB is useful to infer infection phase, and thus to speed clinical decisions in congenital toxoplasmosis management.

Intensity modulated radiation therapy (IMRT) has achieved widespread use for prostate cancer; however, in relation to this use, outcomes studies are still relatively sparse. We report a single-institutional experience in outcomes analysis with the use of IMRT for the primary management of prostate cancer. One hundred thirty consecutive patients with adenocarcinoma of the prostate were treated at a single institution using IMRT with curative intent. Thirty-six (28%) patients were classified as low-risk, 69 (53%) as intermediate-risk, and 25 (19%) as high-risk. The median dose prescription was 76 Gy to the planning target volume. Sixty-five (50%) patients received androgen deprivation therapy (ADT) for a median 4 months, starting 2 months prior to IMRT. Biochemical failure was defined as PSA < post-treatment nadir+2. Gastrointestinal (GI) and Genitourinary (GU) toxicity were defined by RTOG criteria. Median follow-up was 53 months. By NCCN risk category, 4-year biochemical control was 97%, 94%, and 87% for low, intermediate, and high-risk patients, respectively. Among disease factors, multivariable analysis demonstrated the strongest association between biochemical control and Gleason score < or =6 (p=0.0371). Therapy was well tolerated with no Grade 4 toxicity and limited grade 3 GI or GU toxicity. Acute Grade 3+ GI and GU toxicity rates were 0% and 2%, and maximal late Grade 3+ GI and GU toxicity rates were 5% and 6%, respectively. Late rectal toxicity was associated with higher volumes of RT to the rectum. By last follow-up late Grade 3+ toxicity was 2% for both GI and GU systems. In conclusion, patients treated with IMRT for prostate cancer have excellent rates of biochemical control and low rates of severe toxicity of treatment.

An M13 phage random 12-mers peptide library was used to screen cathepsin L mimotopes of Fasciola hepatica and to evaluate their immunogenicity in sheep. Seven clones showed positive reactivity to a rabbit anti-cathepsin L1/L2 antiserum in ELISA, and their amino acid sequences deduced by DNA sequencing were tentatively mapped on the protein. Twenty sheep were randomly allocated into 4 groups of 5 animals each, for immunization with 1x10(14) phage particles of clones 1, 20, a mixture of 7 clones and PBS, without adjuvant at the beginning, and 4 weeks later. All groups were challenged with 300 metacercariae at week 6 and slaughtered 16 weeks later. The mean worm burdens after challenge were reduced by 47.61% and 33.91% in sheep vaccinated with clones 1 and 20, respectively; no effect was observed in animals inoculated with the clone mixture. Also, a significant reduction in worm size and burden was observed for those sheep immunized with clone 1. Animals receiving clone 20, showed a significant reduction in egg output. Immunization induced a reduction of egg viability ranging from 58.92 to 82.11%. Furthermore, vaccinated animals produced clone-specific antibodies which were boosted after challenge with metacercariae of F. hepatica.

Infection of ewes by Toxoplasma gondii may induce abortions, thus it has consequences for sheep production. Colima, Western State of Mexico, has favourable climatic conditions for transmission and both domestic and wild cats live there. The aim of this study was to determine the frequency of specific antibodies in sheep from the coast, a mountain, and a hill of Colima. Serum samples from 351 sheep were tested by a previously standardized indirect ELISA. The frequency of infection was estimated and the farm location and flock size, as well as the animals' age and sex were analysed as risk factors for toxoplasmosis. The frequency of antibodies depended on the altitude, being higher at sea level than at 1200 metres above sea level (OR=3.77, 95% CI=1.79-7.94, P<0.0001), and the size of the flock, being higher in the large ones (OR=2.23, 95% CI=1.35-3.71, P=0.002). Older animals were more frequently positive and with a stronger response than young ones (OR=1.77, 95% CI=1.07-2.93, P=0.016). No differences were observed between male and female sheep. In conclusion, toxoplasmosis is present in sheep of Colima with variations related to altitude, flock size and age.

An indirect ELISA and an immunoblot were standardized to detect anti-Toxoplasma gondii antibodies in sheep, and were compared with a commercial ELISA as reference. Sensitivity was 92.0 and 96.0%, and specificity 88.0 and 75.0%, respectively. Then the serum samples of 103 sheep on a ranch located in the Eastern region of Mexico were analysed. A frequency ranging from 77 to 84% was observed, with a heterogeneous pattern among the animals by immunoblotting. Ten months later 56 sheep were sampled and tested again. Six animals became negative while 1 case was negative the first time and strongly positive 10 months later. Considering this to be a new case we calculated an incidence rate of 2.1% (CI 95% 0.6-4.8%). IgG avidity ELISA was performed on 36 positive samples, 33 being of high-avidity at both times; slight increases in 2 samples and conservation of low-avidity in 1 sheep were also observed. Higher prevalence rates of toxoplasmosis in a moist warm compared to a cold atmosphere was attributed to the long viability of T. gondii oocysts. This may explain the high frequency of T. gondii in this region, which apparently has favourable climatic conditions for the transmission of this protozoan, besides the presence of both domestic and wild cats.

The apicomplexan parasite Toxoplasma gondii is remarkable in several aspects, since it is a protozoan that infects most nucleated cells in many warm-blooded animals, worldwide. Although the cellular immune response against T. gondii is critical for infection control, antibodies may either enhance or block protective mechanisms, and even mediate immunological damage, directly or indirectly. Since cytokines regulate the class/subclass switch, antibodies may also be the biomarkers of protective or pathological cellular immune events. There is a scientific and clinical interest in the presence of natural and autoreactive antibodies, as well as in the 'chronic' immunoglobulin M (IgM) response and the post-treatment 'rebound'. Another interesting aspect is compartmentalization; certain immunoglobulins may uniquely be found in specific host fluids. Local synthesis has been demonstrated, but antibodies may also traverse several cell layers, like the blood-brain and haemato-ocular barriers, and the placenta. In some instances, Fc receptors (FcRs) facilitate transport and may even have a concentrator effect, which can be related to resistance or pathology. These aspects of the humoral response against T. gondii are reviewed in the present paper.

After damage to the central nervous system (CNS) the body is protected by an adaptive immune response which is directed against myelin-associated proteins. Active immunization with nonpathogenic derivatives of CNS-associated peptides (DCAP) reduces the degeneration of neurons and promotes motor recovery after spinal cord injury (SCI) in rats. In order to improve even more the neurological outcome obtained with this therapy, either a combination of DCAP immunization plus glutathione monoethyl ester (GSHE) or a double DCAP immunization were performed. GSHE is a cell-permeant derivative of glutathione, a potent antioxidant agent that significantly inhibits lipid peroxidation after SCI. After a contusive or compressive SCI, the combination of GSHE + DCAP immunization, induced better motor recovery, a higher number of myelinated axons and better rubrospinal neuron survival than immunization alone. On the other hand, double-DCAP immunization counteracted the protective effect of DCAP therapy. Motor recovery and neuronal survival of double-immunized rats were similar to those observed in control animals (PBS-treated). Further studies revealed that double immunization was not encephalitogenic but inhibited the proliferative response of T-cells specific to the DCAP-immunized peptide. This clonal dysfunction was probably secondary to anergy. GSHE improves the protective effect induced by DCAP immunization while double immunization, reverts it.

Study Design: Three independent experiments in a rat model of contusive spinal cord (SC) injury were performed. Two studied the alterations induced by SC injury on some immunological aspects of the T-cell response. The third one evaluated the motor recovery of rats with low-thoracic injuries., Objective: To examine the effect of level, intensity and phase of SC injury on T-cell proliferation and T-cell-dependent antibody response., Setting: Neuroimmunology Department, UIMEN, IMSS-CAMINA Research Center., Methods: Lymphocyte proliferation and hemagglutination assays were performed. Animals were injured either moderately or severely at T1 or T12 SC segments. Analysis of peripheral T-cell proliferation in response to mitogens and to myelin basic protein (MBP), as well as of antibody production against a T-dependent antigen, was performed at acute, subacute and chronic phases., Results: A significant decrease of both response to mitogens and antibody production was found especially during the acute phase and in animals with severe and high (T1)-level injury. Animals with low (T12) and moderate contusions recovered to control levels at the chronic phase. An autoimmune reaction against MBP was observed only in animals with severe contusion at low level., Conclusions: The intensity, level and phase of SC injury differentially alter the function of T cells. These results will allow a better interpretation of studies directed to elucidate the role of T lymphocytes in various processes developed after SC injury.

To compare late genitourinary (GU) and gastrointestinal (GI) toxicity of radiotherapy (RT) to localized fields for prostate cancer delivered using intensity-modulated RT (IMRT) versus conventional RT (ConvRT). The records of 461 patients were reviewed; 355 patients received IMRT and 106 received ConvRT. Late GU and GI toxicity were compared. Late GU toxicity rates were not significantly different (P=0.166); however, late GI toxicity rates were lower with IMRT (P=0.001). Regression analyses demonstrated that only IMRT use (P=0.006) predicted reduction in late GI toxicity but no factors correlated with late GU toxicity. IMRT did not influence late GU toxicity but was associated with a reduction of late GI toxicity over ConvRT.

A 7-year-old Mexican boy with end-stage cirrhosis underwent liver transplantation and was maintained with cyclosporine and prednisolone. No specific data about Toxoplasma gondii or cytomegalovirus (CMV) infections in the cadaver donor were available. The recipient was seronegative for Toxoplasma, but CMV-IgG positive before transplantation. Ganciclovir was administered for prophylaxis during 3 months, but 5 months later he presented with icterus and increased transaminases. Acute transplant rejection was ruled out by biopsy. A seroconversion for T. gondii IgM and IgG and a small increase in CMV-IgM antibodies were observed, although the CMV-polymerase chain reaction (PCR) was negative. Ganciclovir was re-started, and the patient improved, but 6 months later he relapsed, and chorioretinitis lesions compatible both with T. gondii and CMV infections appeared. Pyrimethamine, sulfadiazine, folinic acid, and ganciclovir were administered. The boy showed favorable clinical improvement and remained stable for 12 months. Then, new retinal CMV lesions appeared in both eyes and the PCR for CMV became positive; therefore, the patient received a new regimen of ganciclovir, and clinically improved. From these data we concluded that the child presented a reactivation of CMV and a primary infection with T. gondii after transplantation.

Humidity and temperature have been considered important factors affecting the infectivity of Fasciola hepatica to its molluscan host. One hundred and thirty laboratory-reared Lymnaea humilis were exposed for 4 hr to the miracidia of F. hepatica over a pH range from 4.0 to 10.0, and their rates of survival were compared with 130 similarly treated but unexposed control snails. All control snails died within 24 hr at pH 4.0, but they showed better survival at pH 5.0-10.0. Their sensitivity to solutions with high and low pH, however, was increased if kept in the presence of F. hepatica miracidia. Snails exposed at pH 5.0 died within 24 hr, whereas most other pHs also affected survival such that by day 18 only those snails exposed at pH 7.2 remained alive. The increased sensitivity of the snails to pH could be explained by a damage-mediated release of parasite enzymes, because infectivity was highest at pHs associated with the lowest host mortality.

Antibodies to Toxoplasma gondii were determined by indirect enzyme-linked immunosorbent assay in serum samples from domestic rabbits from 3 rabbit farms in Mexico. Antibodies to T. gondii were found in 77 (26.9%) of 286 animals. On the farm with the higher rearing standards, the seroprevalence was 18.7%, whereas on the farm with medium standards and another managed by a family, seroprevalence was 39.7 and 33.3%, respectively. This report is the first report concerning the prevalence of antibodies to T. gondii in rabbits from Mexico. Although the prevalence found in the present study is within the range reported for other countries, 2 of the farms revealed a relatively high prevalence, which was probably associated with the presence of cats inside rabbit houses.

A 19-month study on the prevalence of fasciolosis in 30 naturally infected cows, the presence of infected and non-infected Lymnaea (Fossaria) humilis snails, and variation in soil temperature and humidity is reported. The prevalence of fasciolosis in cattle declined from around 50% in March to 30% in July, then, it increased from August, reaching a plateau of 100% in November-January, before gradually declining thereafter. A rise in soil humidity and temperature in June and July, respectively, which peaked between August and November was observed. In July, L. (F.) humilis snails appeared, but the infection could only be found in these in August and November. The number of infected snails did not reflect the infestation rate in cows, even though the infestation kinetics in both hosts behaved as predicted from the life cycle of the parasite.

The Taenia solium taeniasis/cysticercosis complex is an important public-health problem in several countries, where many epileptic seizures appear to be associated with neurocysticercosis. As few data on this problem in Nicaragua exist, the seroprevalence of antibodies reacting with antigens from T. solium cysticerci was investigated among 88 Nicaraguan epileptics (45 males and 43 females, aged 6-53 years). In questionnaire-based interviews, each adult subject and a caregiver of each child investigated were asked about potential risk factors for taeniasis/cysticercosis. When a serum sample from each subject was then checked for anti-cysticercus antibodies, 8.0% of the subjects were found seropositive by ELISA and 14.8% by western blotting. Five samples (all from individuals who had been epileptic for > 5 years) were positive in both tests. When the level of association between each potential risk factor and seropositivity (in ELISA or by blotting) was evaluated, the only statistically significant association detected was that between a positive ELISA and the subject living in a household where pigs were raised (odds ratio = 5.18; 95% confidence interval = 0.8-41.6; P = 0.05). The bands most frequently recognized in the western blots (of 50, 42-39, 24 and 14 kDa) were those previously reported. The results indicate that, in the city of Léon, cysticercosis may be endemic and the cause of a significant proportion of the epilepsy recorded.

T cell mediated response is involved in a protective immune response against experimental cysticercosis conferred by immunization with Taenia solium paramyosin (TPmy) to BALB/c mice. In this study, we analysed the TPmy amino acid sequence for predicted CD4+ T cells epitopes. Five different regions of this protein showed that the residues anchor to bind the I-Ad molecule, synthetic peptides containing these epitopes were evaluated for their ability to induce lymphoproliferative responses of spleen cells from TPmy immunized mice. Among them, Tp176 (amino acids 176-192 sequence DDLQRQMADANSAKSRL) was the immunodominant T cell epitope of TPmy. Delineation of this epitope should facilitate analysis of the role of CD4+ T cell response in experimental cysticercosis.

Taenia solium paramyosin is an immunodominant antigen in human and porcine cysticercosis that has shown promise as a vaccine candidate against schistosomiasis and some filariasis. There are few studies to identify the immunologically relevant regions of paramyosin. In this work, we characterize the humoral and cellular response of neurocysticercotic patients against T. solium paramyosin. Western blots using different recombinant fragments of T. solium paramyosin, showed that the sera from neurocysticercotic patients were strongly reactive against the carboxyl end region, with poor recognition of the central and amino regions. In contrast, the cellular immune response of patients did not show preferential recognition of any region of paramyosin.

Quantification of CD4+ T lymphocytes and viral RNA in patients with HIV/AIDS. Levels of progression markers (viral load and CD4+ T lymphocytes) in 410 patients with HIV/AIDS that were in different clinical stages of the disease and under different therapeutic schemes were quantified. The objective was to determine the correlation between values of progression markers and clinical stage of the patients. Commercial methodologies for the quantification of lymphocytes, subpopulations and circulating viral RNA were used. Results indicate that there was a correlation between low CD4+ values and high viral load in patients with antiretroviral treatment but not in patients without treatment. Furthermore, analysis of 1,208 samples processed during 1999 showed that 46% of the patients had less than 200 CD4+ T lymphocytes/mL blood and more than 500 copies of circulating viral RNA. Implications of these results in public health in Mexico are discussed.

Neurocysticercosis is the most common helminthic infection of the CNS but its diagnosis remains difficult. Clinical manifestations are nonspecific, most neuroimaging findings are not pathognomonic, and some serologic tests have low sensitivity and specificity. The authors provide diagnostic criteria for neurocysticercosis based on objective clinical, imaging, immunologic, and epidemiologic data. These include four categories of criteria stratified on the basis of their diagnostic strength, including the following: 1) absolute--histologic demonstration of the parasite from biopsy of a brain or spinal cord lesion, cystic lesions showing the scolex on CT or MRI, and direct visualization of subretinal parasites by funduscopic examination; 2) major--lesions highly suggestive of neurocysticercosis on neuroimaging studies, positive serum enzyme-linked immunoelectrotransfer blot for the detection of anticysticercal antibodies, resolution of intracranial cystic lesions after therapy with albendazole or praziquantel, and spontaneous resolution of small single enhancing lesions; 3) minor--lesions compatible with neurocysticercosis on neuroimaging studies, clinical manifestations suggestive of neurocysticercosis, positive CSF enzyme-linked immunosorbent assay for detection of anticysticercal antibodies or cysticercal antigens, and cysticercosis outside the CNS; and 4) epidemiologic--evidence of a household contact with Taenia solium infection, individuals coming from or living in an area where cysticercosis is endemic, and history of frequent travel to disease-endemic areas. Interpretation of these criteria permits two degrees of diagnostic certainty: 1) definitive diagnosis, in patients who have one absolute criterion or in those who have two major plus one minor and one epidemiologic criterion; and 2) probable diagnosis, in patients who have one major plus two minor criteria, in those who have one major plus one minor and one epidemiologic criterion, and in those who have three minor plus one epidemiologic criterion.

Some reports have suggested that human neurocysticercosis (NCC) induces immunosuppression. To test this hypothesis, we performed a study on active cases of NCC who had not received cestocidal or immunosuppressive treatments. We examined blood counts and specific T cell markers (CD3, CD4, and CD8) by flow cytometry and found no differences between patients with NCC and healthy individuals. Both groups responded to concanavalin A (Con A), and patients with NCC responded more to a parasite crude antigen than uninfected individuals. Peripheral blood mononuclear cells were examined for interleukin (IL)-2, interferon-gamma, IL-10, and IL-4 mRNA. Regardless of infection status, more than 60% of individuals synthesized IL-2 mRNA and, less frequently, the other cytokines. These data suggest that immunosuppression does not occur in NCC patients.

Different assays to detect antigens of Trichinella spiralis during current infection have been standardized, although sensitivity values have been the main limitation to use them as routine diagnostic test. We report the production and use of monoclonal antibodies (MAbs) against the new born larvae which recognize both somatic and metabolic antigens from adult and muscular larvae (ML). We used two IgG3 MAbs (4B1, 4B2) and two IgG2a (2D3, 2D4) to detect antigens during experimental infection. All MAbs detect fecal antigens starting the second to third week post infection (wpi), although it was less clear with MAbs 4B1 and 4B2, while circulating antigens were detected from third to fourth wpi. Thus, the recognition of shared antigens among T. spiralis developmental stages can be used for early diagnosis of trichinellosis.

With the aim to study the magnitude of infection by the metacestode of Taenia solium in a population of epileptic patients in the arid region of Bahia, Northeastern Brazil, we examined 200 consecutive cases who attended an ambulatory clinic in the disctrict of Mulungu do Morro. Sixty-six of the patients had a diagnosis of epilepsy. From them 10 (15.2%) presented antibodies against a specific fraction of antigens in Western blot, and 4 (6.0%) had circulating parasite products, as tested by capture ELISA. Only 1 case was positive for antibodies and antigens. We found that the frequency of seropositivity was related to the time without epileptic seizure. We conclude that cysticercosis is endemic in the region of Mulungu do Morro and that it is related to a benign form of epilepsy.

To compare the effectiveness of cyclosporin-A (CsA) with methylprednisolone (MP) or a combination of both upon inhibition of lipid peroxidation (LP) after spinal cord (SC) injury, rats were treated with either CsA, MP, CSA+MP or vehicle starting 1 h after SC contusion at T9 level. LP was assessed 24h after injury by the lipid fluorescent product formation method. The survival rate was also evaluated in other series of rats by the Kaplan-Meier curves. Lipid peroxidation was similarly inhibited in rats treated with CsA, MP, or CSA+MP (p>0.05). Animals receiving MP (alone or combined with CsA) showed the poorest surviving rate. LP was inhibited by CsA to the same extent as by MP but without the lethal effect of the latter.

Hydatid disease of the liver remains an important and challenging problem in rural areas; although, surgery is considered the treatment of choice, percutaneous treatment of hydatid cysts is relatively new, and the data related to it are limited. The purpose of the study was to present the results of percutaneous treatment of liver hydatid cysts. Thirty-four patients (13 male and 21 female), ranging in age between 14 and 80 years, with 55 liver hydatid cysts underwent percutaneous treatment with albendazole prophylaxis. Cysts were treated with a one-stage procedure that consisted of puncture of the cysts under guidance with computed tomography, aspiration of fluid, injection of hypertonic saline solution as scolicidal agent and reaspiration. Follow-up examinations showed progressive reduction and solidification of the cysts. The mean reduction in volume was 72%. No mortality, abdominal dissemination, or tract seeding occurred. Minor complications were urticaria with pruritus in two patients. One patient had a subcapusular hematoma without problem. Hospitalization courses varied from ambulatory procedures to 15 days of in-patient, mean hospital stay was 1.82 days. The results of percutaneous liver hydatid cyst treatment, indicating that the procedure is efficient and safe and offers complete cure in selected patients with a short hospitalization and that this technique should be considered an alternative to surgery.

In order to determine if a specific response is induced after spinal cord injury, we performed a kinetic search for IgG antibodies against various spinal cord antigenic preparations in a rat contusion model. Even though spinal cord injured animals showed two reactive bands, these could be originated by the reaction of natural antibodies, since they were also observed before lesion. Thus, these antibodies would not be of relevance in the pathogenic events of spinal cord injury in this rat model. Our findings do not demonstrate the existence of a specific IgG response against spinal cord constituents after injury.

To determine markers of Taenia solium transmission and risk factors in an urban community, we studied 1,000 soldiers from a military camp in Mexico City and their relatives. Serum samples were used to detect antigens and antibodies and fecal specimens were examined for Taenia coproantigens and helminth eggs. Prevalences of 12.2% and 5.8% for cysticercosis were found among soldiers and their relatives, respectively. Taeniasis was found in 0.5% and none of the groups, respectively. Relatives of soldiers positive for cysticercosis and taeniasis markers ate more pork from street stores than restaurants or markets compared with relatives of soldiers without these indicators of infection. Also, 12.0% of the relatives of positive soldiers had a history of expelling tapeworm proglottids in the feces in contrast to 3.7% of the family members of the control group. Prevalence values and risk factors in this urban population are similar to those of previous studies performed in rural populations.

Percutaneous puncture-aspiration-injection-reaspiration (PAIR) of hydatid liver cysts, was performed in 38 patients 14-80 years old, with a total of 60 liver hydatid cysts. After aspiration under computed tomography guidance, hypertonic saline was injected into the cystic cavities of patients as a scolecidal agent. No major complications were associated with the procedures. In the follow-up period of 18 months, control CT scans of 35 cysts revealed a gradual decrease in cyst size with a mean volume reduction of 66%. Complications included two cases of urticaria, one case of anaphylaxis and one subcapsular hematoma. No mortality occurred. It is concluded that percutaneous aspiration and hypertonic saline injection for liver hydatid cysts appears to be an effective form of treatment and may eventually prove to be an alternative to surgical intervention.

Besides its immunosuppressive/anti-inflammatory activity, cyclosporin-A (CsA) may protect damaged tissues from lipid peroxidation (LP) by free radicals. To determine the effect of CsA on LP spinal cord (SC) injury, Wistar rats were treated with either vehicle or CsA (2.5 mg/kg per 12 h i.p.) 1, 2, 6 or 12 h after SC trauma by T8-T9 spinal cord contusion, analyzing LP 24 h after injury at the lesion site by the lipid fluorescent products formation method. CsA significantly diminished LP to levels below control values after contusion (P < 0.05). The greater inhibition was observed when CsA was given during the first 6 h after injury, furthermore, animals showed a significant clinical improvement. Results show that CsA may be beneficial to injured tissue by inhibiting the levels of LP.

Human neurocysticercosis is an important parasitic disease in developing countries. Most epidemiological studies on the disease have used antibody-based assays that allow the detection of transmission 'hot spots' and the identification of the main risk factors for transmission. However, such assays have low predictive value in the detection of active cases of neurocysticercosis. The screening potential of the most commonly used antibody-detection technique, the electroimmunotransfer blot assay (EITB), has now been compared with an antigen-capture assay, in an endemic region of Mexico. The subjects were 68 patients with late-onset epilepsy, 35 cases of taeniasis and a randomly selected, control group of 133 individuals from the same region. Parasite-specific antibodies and antigens were more common among the epileptics and taeniasis cases than among the controls. The antigens appeared to be associated with late-onset epilepsy and the antibodies with the presence of subcutaneous nodules. The sensitivities of both tests, to detect epilepsy or taeniasis, were low, but the specificity and the positive predictive value of the antigen-capture assay was high when used with the epileptics. As late-onset epilepsy and neurocysticercosis seem to be associated in endemic regions, antigen-capture assays are probably the most reliable method of detecting active cases of neurocysticercosis in epidemiological studies.