Transcriptome profiling of macrophages persistently infected with human respiratory syncytial virus and effect of recombinant Taenia solium calreticulin on immune-related genes.

Introduction: Human respiratory syncytial virus (hRSV) is a main cause of bronchiolitis in infants and its persistence has been described in immunocompromised subjects. However, limited evidence has been reported on the gene expression triggered by the hRSV and the effect of recombinant Taenia solium-derived calreticulin (rTsCRT). Methods: Using a comprehensive microarray approach, we analyzed the transcriptome profile of a macrophage cell line that has supported hRSV persistence for over 150 passages. We compared the gene expression of persistently infected and non-infected macrophages. We also evaluated the effect of rTsCRT on hRSV-infected macrophage gene transcription, as well as on cytokine production and number of copies of the persistent hRSV genome. Results: Our analysis showed that hRSV long-term virus infection significantly alters mRNA expression of antiviral, inflammatory, as well as arginine and lipid metabolism-associated genes, revealing a transcriptional signature that suggests a mixed M1/M2 phenotype. The resulting host-virus equilibrium allows for the regulation of viral replication, while evading the antiviral and proinflammatory responses. Interestingly, rTsCRT stimulus upregulated Tnfα, Il6 and Nos2 mRNA. We found increased levels of both proinflammatory cytokines and nitrite levels in the conditioned media of persistent macrophages treated with rTsCRT. This increase was associated with a significant reduction in viral genome copies. Discussion: hRSV persistently infected macrophages retain responsiveness to external stimuli and demonstrate that the profound changes induced by viral persistence are potentially reversible. Our observations contribute to the understanding of the mechanisms related to hRSV persistence in macrophages and have implications for the development of targeted therapies to eliminate persistent infections or reduce the negative effects related with chronic inflammatory diseases associated with hRSV infection.

Splenocytes and thymocytes migration patterns between lymphoid organs in pregnancy.

Background: Cell migration is essential for the immune system and is frequently analyzed in adult non-pregnant animals but poorly explored in pregnant animals. However, a physiologic increased size in the spleen and periaortic lymph nodes had been reported in pregnant mice. Methods: Using a mouse model, we transferred PKH26-stained thymocytes and splenocytes from pregnant or non-pregnant animals to receptor mice in the presence or absence of pregnancy. Percentage of PKH-26 cells and Mean Fluorescence Intensity were calculated. Non-parametric ANOVA analysis was performed. Results: We detected that the percentage of PKH26+ thymocytes in the spleen, lymph nodes, and peripheral blood is higher in females than in males (p = 0.039). Our results showed a similar frequency of thymocytes and splenocytes from pregnant and non-pregnant mice located in receptor lymphoid organs (p > 0.05). Also, the location of marked cells was similar during the perinatal period (p > 0.05). Conclusions: The mobility of thymocytes and splenocytes in pregnant and non-pregnant mice is similar. Therefore, we suggest that the larger size of the spleen and periaortic lymph nodes noted previously in pregnant mice could result from the retention of leukocytes in the secondary lymphoid organs.

Fc-gamma receptor expression and cytokine responses to intravenous human immunoglobulin in whole blood from non-pregnant and pregnant women and newborns.

BACKGROUND: Intravenous immunoglobulin (IVIG) can suppress the inflammatory response in adults, but its role in pregnant women and newborns is poorly studied. While the adult immune system is considered mature, it is immature in neonates and suppressed in pregnancy. Since the immune response differs in these 3 groups, the use of IVIG could differentially modulate the immune response. OBJECTIVES: We aimed to explore the effect of IVIG on myeloid blood cells from non-pregnant women, pregnant women and newborns. MATERIAL AND METHODS: Whole blood from healthy donors was incubated with lipopolysaccharide (LPS) and/or IVIG. After 0 h, 24 h and 48 h of culture, Fc-gamma receptor (CD16, CD32 and CD64) expression, monocyte and neutrophil bacterial phagocytosis, and cytokine and chemokine concentrations were determined in the supernatant. RESULTS: The baseline expression of monocyte CD16 was higher in newborns than in adult women, but the expression of CD32 and CD64 was similar between groups. Furthermore, LPS and IVIG stimulation, together or separately, did not change Fc-gamma receptor expression in monocytes or neutrophils and did not modify their phagocytosis capacity. On the other hand, IVIG did not downregulate the proinflammatory cytokine response induced by LPS in any group. Interestingly, IVIG induced a strong interleukin 8 (IL-8) response in neonates but not in non-pregnant or pregnant women. CONCLUSIONS: Our results show that IVIG did not induce changes in Fc-gamma receptor expression, phagocytic ability, or the cytokine response to LPS in blood cells from neonates, non-pregnant or pregnant women. However, IVIG induced a strong IL-8 response in neonates that could improve immunity.

An Overview on Research in a University Hospital, Using a Payback Framework Categorization Approach.

BACKGROUND AND OBJECTIVE: Little information is available on how to assess the impact of research studies conducted in government hospitals in Latin America and specifically in Mexico. We aimed to determine the returns on investment of the research projects that were carried out in the Hospital General "Dr. Manuel Gea Gonzalez" (HGMGG), a general university hospital located in Mexico City, using a categorization model. METHODS: We conducted a study including bibliometric analyses of publications associated with all research studies performed during the period 2016-2019 in the HGMGG and investigator interviews, according to the payback framework categorization model. RESULTS: All studies analyzed had a positive impact based on outcomes in 5 "payback categories": (1) knowledge; (2) research targeting, capacity building, and absorption; (3) policy and product development; (4) health benefits; and (5) broader economic benefits. CONCLUSIONS: To date, it has not been possible to establish a set of indicators that show the results of the investigations carried out by medical specialists in training, who carry out the bulk of medical care in general hospitals and in the National Institutes of Health in Mexico. We identified, in the 5 categories of the payback framework model, different areas of opportunity to improve the benefits of the hospital's medical services through the development of scientific research projects.

Elevated Levels of Cytotoxicity, Cytokines, and Anti-SARS-CoV-2 Antibodies in Mild Cases of COVID-19.

Current evidence shows higher production of cytokines and antibodies against severe acute respiratory coronavirus 2 (SARS-CoV-2) in severe and critical cases of Coronavirus Disease 2019 (COVID-19) in comparison with patients with moderate or mild disease. A recent hypothesis proposes an important role of genotoxicity and cytotoxicity in the induction of the cytokine storm observed in some patients at later stages of the disease. Interestingly, in this study, we report significantly higher levels of interleukin (IL)-1ß, IL-6, MCP-1, and IL-4 cytokines in mild COVID-19 patients versus severe cases, as well as a high frequency of karyorrhexis (median [Me] = 364 vs. 20 cells) and karyolysis (Me = 266 vs. 52 cells) in the mucosal epithelial cells of both groups of patients compared with uninfected individuals. Although we observed higher levels of anti-SARS-CoV-2 IgM and IgG antibodies in COVID-19 patients, IgM antibodies were significantly higher only in mild cases, for the N and the S viral antigens. High levels of IgG antibodies were observed in both mild and severe cases. Our results showed elevated concentrations of proinflammatory and anti-inflammatory cytokines in mild cases, which may reflect an active innate immune response and could be related to the higher IgM and IgG antibody levels found in those patients. In addition, we found that SARS-CoV-2 infection induces cytotoxic damage in the oral mucosa, highlighting the importance of studying the genotoxic and cytotoxic events induced by infection and its role in the pathophysiology of COVID-19.

Why are tapeworm carriers so difficult to find? A methodological proposal for their search and recovery.

Neurocysticercosis, caused by the larval stage of Taenia solium, is a life-threatening condition and the most severe form of the disease. Yet, despite being a required link in the parasite life cycle, tapeworm carriers are rarely reported. This study is aimed to find and evaluate T. solium carriers, describing some characteristics of these patients and the treatment. Taeniasis cases were searched for in various Mexican states from 1983 to 2016. Previous informed consent, tapeworm-carrier patients were administered with niclosamide and a saline purge. Parasite specimens were recovered and identified, both morphologically and by PCR. From 117 treated patients, Taenia sp. specimens were obtained from 46 subjects (47.8%). From these, complete parasites were recovered from 42 (90.5%), and only detached proglottids from 4 patients. Cases were more frequent in Morelos, Chiapas, and Guerrero. More than one adult cestode was recovered from 4 patients (9.5%). To improve treatment efficacy and adherence, the drug was administered in late afternoon, resulting a high recovery yield of complete parasites (90.5%). The success rate of deworming campaigns in areas of Mexico and the world that are endemic for Taenia sp. could be improved by administering the treatment at times that do not interfere with the patients' daily activities, and national health authorities could apply this simple strategy to help eradication efforts in endemic areas. The detection of carriers will only be possible through the coordinated efforts of public and private health services, a better education of the general population to improve self-detection, and adequate, personalized diagnostic procedures for suspect cases.

The many faces of parasite calreticulin.

Calreticulin from parasites and its vertebrate hosts share ~50% identity and many of its functions are equally conserved. However, the existing amino acid differences can affect its biological performance. Calreticulin plays an important role in Ca2+ homeostasis and as a chaperone involved in the correct folding of proteins within the endoplasmic reticulum. Outside the endoplasmic reticulum, calreticulin is involved in several immunological functions such as complement inhibition, enhancement of efferocytosis, and immune upregulation or inhibition. Several parasite calreticulins have been shown to limit immune responses and promote infectivity, while others are strong immunogens and have been used for the development of potential vaccines that limit parasite growth. Furthermore, calreticulin is essential in the dialogue between parasites and hosts, inducing Th1, Th2 or regulatory responses in a species-specific manner. In addition, calreticulin participates as initiator of endoplasmic reticulum stress in tumor cells and promotion of immunogenic cell death and removal by macrophages. Direct anti-tumoral activity has also been reported. The highly immunogenic and pleiotropic nature of parasite calreticulins, either as positive or negative regulators of the immune response, render these proteins as valuable tools to modulate immunopathologies and autoimmune disorders, as well as a potential treatment of neoplasms. Moreover, the disparities in the amino acid composition of parasite calreticulins might provide subtle variations in the mechanisms of action that could provide advantages as therapeutic tools. Here, we review the immunological roles of parasite calreticulins and discuss possible beneficial applications.

Neurocysticercosis in Latin America: Current epidemiological situation based on official statistics from four countries.

BACKGROUND: Neurocysticercosis (NC) is one of the major parasitic diseases affecting the central nervous system and is endemic in much of Asia, sub-Saharan Africa, and Latin America. Its epidemiology is difficult to assess, although official registries are available in Brazil, Colombia, Ecuador, and Mexico. METHODOLOGY/PRINCIPAL FINDINGS: Using official statistics, we assessed trends in NC hospitalization rates during 1998-2019 in Brazil and Ecuador, during 2004-2019 in Mexico, and during 2009-2019 in Colombia. We also assessed the trend in NC mortality in Brazil (1998-2019), the trend in hospitalizations for NC in a Mexican tertiary-level hospital (Instituto Nacional de Neurología y Neurocirugía [INNN]; 1995-2019), and in Mexican primary care ambulatory clinics (1995-2019). Associations between NC hospitalization rates and the human development index (HDI) were also examined. In Brazil, Ecuador, and Mexico, statistically significant decreases in NC hospitalization rates were observed. In Mexico, a significant increase in the age of patients at INNN was observed, suggesting a decreasing incidence of recent infection. Conversely, a significant increase in NC hospitalization rate was observed in Colombia. HDI was not significantly associated with NC hospitalization rates when adjusting for time. CONCLUSIONS: The downward trends in NC cases in Brazil, Ecuador, and Mexico are encouraging, especially in the context of the PAHO/WHO plan of action to eliminate neglected tropical diseases from the region. On the other hand, in Colombia, the increased NC hospitalization rate is concerning and needs further evaluation so that the authorities can take specific measures. These results should encourage health authorities in other endemic countries to establish a system of official registries to identify where the need for a control program is most urgent. However, it is also important to remember that NC persists, although less frequently in some Latin American countries, and efforts to achieve its control must continue.

Identification of α-L-fucosidase (ALFuc) of Blastocystis sp. subtypes ST1, ST2 and ST3.

Blastocystis sp. is a common intestinal microorganism. The α-L-fucosidase (ALFuc) is an enzyme long associated with the colonization of the gut microbiota. However, this enzyme has not been experimentally identified in Blastocystis cultures. The objective of the present study was to identify ALFuc in supernatants of axenic cultures of Blastocystis subtype (ST)1 ATCC-50177 and ATCC-50610 and to compare predicted ALFuc proteins of alfuc genes in sequenced STs1-3 isolates in human Blastocystis carriers. Excretion/secretion (Es/p) and cell lysate proteins were obtained by processing Blastocystis ATCC cultures and submitting them to SDS-PAGE and immunoblotting. In addition, 18 fecal samples from symptomatic Blastocystis human carriers were analyzed by sequencing of amplification products for subtyping. A complete identification of the alfuc gene and phylogenetic analysis were performed. Immunoblotting showed that the amplified band corresponding to ALFuc (~51 kDa) was recognized only in the ES/p. Furthermore, prediction analysis of ALFuc 3D structures revealed that the domain α-L-fucosidase and the GH29 family's catalytic sites were conserved; interestingly, the galactose-binding domain was recognized only in ST1 and ST2. The phylogenetic inferences of ALFuc showed that STs1-3 were clearly identifiable and grouped into specific clusters. Our results show, for the first time through experimental data that ALFuc is a secretion product of Blastocystis sp., which could have a relevant role during intestinal colonization; however, further studies are required to clarify this condition. Furthermore, the alfuc gene is a promising candidate for a phylogenetic marker, as it shows a conserved classification with the SSU-rDNA gene.

Identification of α-L-fucosidase (ALFuc) of Blastocystis sp. subtypes ST1, ST2 and ST3

ABSTRACT Blastocystis sp. is a common intestinal microorganism. The α-L-fucosidase (ALFuc) is an enzyme long associated with the colonization of the gut microbiota. However, this enzyme has not been experimentally identified in Blastocystis cultures. The objective of the present study was to identify ALFuc in supernatants of axenic cultures of Blastocystis subtype (ST)1 ATCC-50177 and ATCC-50610 and to compare predicted ALFuc proteins of alfuc genes in sequenced STs1-3 isolates in human Blastocystis carriers. Excretion/secretion (Es/p) and cell lysate proteins were obtained by processing Blastocystis ATCC cultures and submitting them to SDS-PAGE and immunoblotting. In addition, 18 fecal samples from symptomatic Blastocystis human carriers were analyzed by sequencing of amplification products for subtyping. A complete identification of the alfuc gene and phylogenetic analysis were performed. Immunoblotting showed that the amplified band corresponding to ALFuc (~51 kDa) was recognized only in the ES/p. Furthermore, prediction analysis of ALFuc 3D structures revealed that the domain α-L-fucosidase and the GH29 family's catalytic sites were conserved; interestingly, the galactose-binding domain was recognized only in ST1 and ST2. The phylogenetic inferences of ALFuc showed that STs1-3 were clearly identifiable and grouped into specific clusters. Our results show, for the first time through experimental data that ALFuc is a secretion product of Blastocystis sp., which could have a relevant role during intestinal colonization; however, further studies are required to clarify this condition. Furthermore, the alfuc gene is a promising candidate for a phylogenetic marker, as it shows a conserved classification with the SSU-rDNA gene.

Association between cycle threshold (Ct ) values and clinical and laboratory data in inpatients with COVID-19 and asymptomatic health workers.

In-house assays for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), are feasible alternatives, particularly in developing countries. Cycle threshold (Ct ) values obtained by qRT-PCR were compared with clinical and laboratory data from saliva of inpatients with COVID-19 and asymptomatic health workers (AHW) were studied. Saliva specimens from 58 inpatients confirmed by qRT-PCR for SARS-CoV-2 using nasopharyngeal specimens, and 105 AHW were studied by qRT-PCR using three sets of primers for the N (N1, N2, and N3) gene of SARS-CoV-2, according to the CDC Diagnostic Panel protocol, showing a positivity of 88% for inpatients and 8% for AHW. Bivariate analysis revealed an association between Ct < 38.0 values for N2 and mechanical ventilation assistance among patients (p = .013). In addition, values of aspartate-transaminase, lactate dehydrogenase, and ferritin showed significant correlations with Ct values of N1 and N3 genes in inpatients. Therefore, our results show that Ct values correlate with some relevant clinical data for inpatients with COVID-19.

Zoonotic Taenia infections with focus on cysticercosis due to Taenia solium in swine and humans.

Zoonotic taeniasis caused by the adult stage of Taenia solium, Taenia saginata or Taenia asiatica are considered neglected tropical diseases by the World Health Organization. The life cycle of these 3 metazoan species is very similar and includes an intermediate host: pigs in the case of T. solium and T. asiatica, and cattle in the case of T. saginata. By eating meat (pork/T. solium, T. asiatica; beef/T. saginata) containing live cysticerci, humans develop taeniasis, which is practically asymptomatic but is the main risk factor for intermediate hosts to become infected. T. saginata causes bovine cysticercosis, while T. solium and T. asiatica cause swine cysticercosis, of veterinary and economic importance. T. solium cysticerci cause neurological disease in humans: neurocysticercosis. Cysticerci develop after ingesting microscopic eggs released from a human tapeworm carrier. Here we describe the life stages of the parasites, diagnosis, pathogenesis, symptomatology of neurocysticercosis, and prevention and control measures. Highlighting the need to validate diagnostic tools, treatments and vaccination in endemic areas, with the challenge of addressing the most vulnerable populations that lack resources. If people understand the transmission route, avoid eating uncooked or insufficiently cooked meat and have adequate hygienic habits, the life cycle of the 3 zoonotic Taenia species may be interrupted. In addition, we describe the growing field of immune response and immunomodulation elicited by the parasites, which may provide essential tools for diagnosis, treatment, control of taeniasis/cysticercosis, as well as for identification of parasite-derived immunomodulators that could aid in the treatment of emerging inflammatory diseases worldwide.

Measuring student satisfaction as the first assessment of the Program of Combined Studies in Medicine, an MD/PhD-like program of the Faculty of Medicine, National Autonomous University of Mexico.

BACKGROUND: The Program of Combined Studies in Medicine (PECEM, by its acronym in Spanish) is a program for simultaneous bachelor and doctorate studies (MD/PhD) that enrolls students who show academic excellence and interest in scientific research. The initial doctoral training comprises seven six-month research stays in different laboratories or clinical or computer areas with different high-quality scientific advisors who provide students with a unique experience for their scientific training. Therefore, satisfaction in this stage is decisive for students' performance and physical and psychological health. The aim of the present study was to administer a questionnaire to measure students' satisfaction with their research experience as a service-product bundle. METHODS: Students answered an online questionnaire that evaluated three dimensions: perceived quality of the advisor, skills development, and infrastructure and support. Several satisfiers were also evaluated: recommendation of the advisor to peers, fulfillment of student expectations and satisfaction with the program. Correlations were calculated using Fisher's exact test. The significance was set at p < 0.05. RESULTS: The high quality of the advisor, skills development and guidance during the stay were satisfiers correlated to the students' recommendation of their advisors to their peers and to the fulfillment of the students' scientific expectations. Conversely, skills development and infrastructure and support were satisfiers for a good to excellent experience as a PECEM student. A lack of direct interaction with the advisor's workgroup was related to dissatisfaction. CONCLUSIONS: The nontangible products of the service, such as a positive interaction between the student, the advisor and the advisor's workgroup as well as support obtained during the research stay, were satisfiers. These data indicate that promoting a fruitful bond between the student and advisor is a priority to ensure the quality of our innovative MD/PhD program.

Influenza A H1N1 Virus 2009 Synthetic Hemagglutinin and Neuraminidase Peptides for Antibody Detection.

BACKGROUND: Influenza serologic diagnosis is mainly based on hemagglutination inhibition and microneutralization methods, both methods require handling living viruses under an enhanced biosafety level. AIM: The current study was performed for developing an ELISA using synthetic peptides to detect influenza A H1N1 virus 2009 specific antibodies in serum and saliva. METHODS: Alignments were made with H1N1 hemagglutinin and neuraminidase (HA and NA, respectively) sequences; only conserved sites were used for antigenicity prediction. Two synthetic peptides were assayed; one of neuraminidase (NA15) and one of hemagglutinin (HA-15) and used in ELISA for detecting IgG and IgA antibodies. A cross-sectional study was performed in three municipalities of Mexico City, using negative samples collected before the 2009 influenza outbreak, samples of people who became ill during the outbreak, and samples of the participants in the epidemiological study with or without symptoms. RESULTS: The determination of serum IgG antibodies with both peptides allowed differentiating between the post outbreak groups with respect to all others. No differences were found in IgA determination in saliva against both peptides. The frequency of positive participants for NA-15 was 9.5 and 8.8% for HA-15 in serum IgG; whereas the frequency of positive participants for NA-15 was 11%, and for HA-15 was 8.6% for saliva IgA. CONCLUSIONS: Synthetic peptides of the neuraminidase and hemagglutinin proteins can be used in ELISA for the determination of IgG and IgA antibodies against the influenza A H1N1 virus 2009.

In Vitro Employment of Recombinant Taenia solium Calreticulin as a Novel Strategy Against Breast and Ovarian Cancer Stem-like Cells.

BACKGROUND AND AIMS: Calreticulin is a chaperone and master regulator of intracellular calcium homeostasis. Several additional functions have been discovered. Human and parasite calreticulin have been shown to suppress mammary tumor growth in vivo. Here, we explored the capacity of recombinant Taenia solium calreticulin (rTsCRT) to modulate cancer cell growth in vitro. METHODS: We used different concentrations of rTsCRT to treat cancer cell lines and analyzed viability and colony formation capacity. We also tested the combination of the IC20 or IC50 doses of rTsCRT and of the chemotherapeutic drug 5-fluorouracil on MCF7 and SKOV3 cell lines. As a control, the non-tumorigenic cell line MCF10-A was employed. The effect of the drug combinations was also assessed in cancer stem-like cells. Additionally, scavenger receptor ligands were employed to identify the role of this receptor in the rTsCRT anti-tumoral effect. RESULTS: rTsCRT has a dose-dependent in vitro anti-tumoral effect, being SKOV3 the most sensitive cell line followed by MCF7. When rTsCRT/5-fluorouracil were used, MCF7 and SKOV3 showed a 60% reduction in cell viability; colony formation capacity was also diminished. Treatment of cancer stem-like cells from MCF7 showed a higher reduction in cell viability, while those from SKOV3 were more sensitive to colony disaggregation. Finally, pharmacological inhibition of the scavenger receptor, abrogated the reduction in viability induced by rTsCRT in both the parental and stem-like cells. CONCLUSION: Our data suggest that rTsCRT alone or in combination with 5-fluorouracil inhibits the growth of breast and ovarian cancer cell lines through its interaction with scavenger receptors.

An Unexpected Case of Lagochilascariasis: Interdisciplinary Management and Use of 12S and 18S rDNA Analysis.

A Mexican 24-year-old male patient was referred to our hospital due to increased left retroauricular volume with skin fistulisation, resembling an infection by the uncommon worm Lagochilascaris minor. The patient was submitted to lateral skull base surgery. No adult worms or eggs were observed during light and scanning electron microscopy analysis, as well as by histopathologic examination of the small piece of removed tissue, only L3 stage larvae of Lagochilascaris spp. were identified. Polymerase chain reaction-sequencing assays were performed using primers for the mitochondrial 12S and the nuclear 18S rDNA gene. DNA of some L minor adults, previously identified, were used as control. The molecular analysis identified the worm as L minor. According to previous reports, lagochilascariasis is a complicated infection that requires an interdisciplinary management by different clinical specialists. This is the first time that 12S and 18S rDNA genes are reported as molecular markers for diagnosis of L minor.

The monetary burden of cysticercosis in Mexico.

BACKGROUND: Taenia solium cysticercosis is a public health and agricultural problem in many low and middle-income countries where health education, sanitation, pig management practices and meat inspection infrastructure are insufficient. Cysticercosis affects both human and animal health and has important economic consequences. Very few studies have been conducted to evaluate the monetary burden of cysticercosis. This study aimed at estimating the 2015 costs associated with cysticercosis in humans and pigs in Mexico. METHODS: The monetary burden of human cysticercosis was estimated based on costs incurred by living with and treating epilepsy and severe chronic headaches associated with neurocysticercosis (NCC). The estimated cost of porcine cysticercosis took into consideration losses due to the reduction in the price of cysticercosis-infected animals. Epidemiologic and economic data were obtained from the published literature, government reports, and setting-specific questionnaires. Latin hypercube sampling methods were employed to sample the distributions of uncertain parameters and to estimate 95% credible regions (95% CRs). All results are reported in 2015 U.S.$. FINDINGS: The overall monetary burden associated with NCC morbidity was estimated at U.S.$215,775,056 (95% CR U.S.$109,309,560 -U.S.$361,924,224), with U.S.$436 (95% CR: U.S.$296 -U.S.$604) lost per patient. If loss of future years of income and productivity due to NCC-associated deaths was included, this value increased by U.S.$54.26 million, assuming that these individuals earned Mexico's median wage salary. An additional U.S.$19,507,171 (95% CR U.S.$5,734,782 -U.S.$35,913,487) was estimated to be lost due to porcine cysticercosis. CONCLUSIONS: This study suggests that T. solium cysticercosis results in considerable monetary losses to Mexico.

Calreticulin in phagocytosis and cancer: opposite roles in immune response outcomes.

Calreticulin (CRT) is a pleiotropic and highly conserved molecule that is mainly localized in the endoplasmic reticulum. Recently, CRT has gained special interest for its functions outside the endoplasmic reticulum where it has immunomodulatory properties. CRT translocation to the cell membrane serves as an "eat me" signal and promotes efferocytosis of apoptotic cells and cancer cell removal with completely opposite outcomes. Efferocytosis results in a silenced immune response and homeostasis, while removal of dying cancer cells brought about by anthracycline treatment, ionizing-irradiation or photodynamic therapy results in immunogenic cell death with activation of the innate and adaptive immune responses. In addition, CRT impacts phagocyte activation and cytokine production. The effects of CRT on cytokine production depend on its conformation, species specificity, degree of oligomerization and/or glycosylation, as well as its cellular localization and the molecular partners involved. The controversial roles of CRT in cancer progression and the possible role of the CALR gene mutations in myeloproliferative neoplasms are also addressed. The release of CRT and its influence on the different cells involved during efferocytosis and immunogenic cell death points to additional roles of CRT besides merely acting as an "eat me" signal during apoptosis. Understanding the contribution of CRT in physiological and pathological processes could give us some insight into the potential of CRT as a therapeutic target.

RNA Purity, Real-Time PCR Sensitivity, and Colon Segment Influence mRNA Relative Expression in Murine Dextran Sodium Sulfate Experimental Colitis.

The dextran sodium sulfate (DSS) model of colitis is widely used as a result of its simplicity and reproducibility and because it mimics clinicopathological disease features. Its effectiveness depends on the mouse strain, DSS MW, and brand. Quantitative RT-PCR (qRT-PCR) is highly sensitive for analyzing cytokine mRNA expression. We analyzed an acute model of DSS treatment in Balb/c mice for the onset of colitis using qRT-PCR for the quantification of a mouse cytokine transcript. We compared differences among 1--and 2-step qRT-PCR for transcript quantification, the effect of multiple concentrations of DSS, and the use of 2 reference genes in 3 portions of the colon. A reliable and sensitive 1-step protocol for qRT-PCR was established with a modified double LiCl precipitation for RNA isolation. The variability of 2 reference genes, ß-actin and eukaryotic elongation factor 2, was compared, and expression of IL-6 was analyzed in 3 segments of the colon. The RNA cleaning protocol prevented inhibition of qRT-PCR by DSS, and RNA loss was minimized. No clinical differences among the different DSS concentrations were seen on d 7, but higher concentrations resulted in the appearance of earlier symptoms. Higher efficiency and sensitivity of the 1-step qRT-PCR reaction using eukaryotic elongation factor 2 were obtained and also less variability. Although expression levels of IL-6 were high in the middle and distal colon, the middle section had consistently less variability in values. Thus, this segment is recommended for future studies. These factors influence the statistical significance of data and need to be considered to get accurate and reliable results and to improve comparisons of the published colitis experiments.

Fate of uptaken host proteins in Taenia solium and Taenia crassiceps cysticerci.

During the study of host-parasite relationships in taeniid parasite diseases, including cysticercosis and hydatidosis, reports have described the presence of host proteins in the cyst fluid and tissue of metacestodes. However, the fate or role of host elements inside the parasite remains barely explored. After the publication of genomes of four cestode species, it became clear that these organisms possess a limited biosynthetic capability. The initial goal of the present study was to determine if uptaken host proteins could be a source of essential amino acids for cysticerci. To track the utilization of uptaken proteins, we added metabolically labeled IgG-3H and GFP-3H to the culture medium of Taenia crassiceps cysticerci. Incorporation of labeled amino acid was evaluated by fluorography in cysticerci extracts. Our results showed that the use of uptaken proteins by cysticerci as a source of amino acids appeared negligible. Exploring alternative fates for the host proteins, proteomic analysis of the protein matrix in calcareous corpuscles was carried out. Since T. crassiceps does not contain calcareous corpuscles, proteomic analyses were performed in corpuscles of Taenia solium cysticerci. Our results demonstrated that host proteins represented approximately 70% of protein content in the calcareous corpuscles. The presence of the two major uptaken host proteins, namely albumin and IgG, was also demonstrated by Western blot in the matrix of corpuscles. Our findings strongly suggested that the uptake and disposal of host proteins involve calcareous corpuscles, expanding the physiological role of these mineral concretions to a far more important level than previously proposed.