Evaluation of the safety and adjuvant effect of a detoxified listeriolysin O mutant on the humoral response to dengue virus antigens.

Listeriolysin O (LLO) has been proposed as a potential carrier or adjuvant molecule in the vaccination field. However, the cytotoxic and pro-apoptotic effects of LLO are the major limitations for this purpose. Here, we have performed a preclinical safety evaluation and characterized a new potential adjuvant application for a non-cytolytic LLO mutant (dtLLO) to enhance and modulate the immune response against the envelope (E) protein from dengue virus. In addition, we have studied the adjuvant effects of dtLLO on human immune cells and the role of membrane cholesterol for the binding and proinflammatory property of the toxoid. Our in-vivo results in the murine model confirmed that dtLLO is a safer molecule than wild-type LLO (wtLLO), with a significantly increased survival rate for mice challenged with dtLLO compared with mice challenged with wtLLO (P < 0·001). Histopathological analysis showed non-toxic effects in key target organs such as brain, heart, liver, spleen, kidney and lung after challenge with dtLLO. In vitro, dtLLO retained the capacity of binding to plasma membrane cholesterol on the surface of murine and human immune cells. Immunization of 6-8-week-old female BALB/c mice with a combination of dtLLO mixed with E protein elicited a robust specific humoral response with isotype diversification of immunoglobulin (Ig)G antibodies (IgG1 and IgG2a). Finally, we demonstrated that cholesterol and lipid raft integrity are required to induce a proinflammatory response by human cells. Taken together, these findings support a potential use of the dtLLO mutant as a safe and effective adjuvant molecule in vaccination.

Non-Conjugated Chitosan-Based Nanoparticles to Proteic Antigens Elicit Similar Humoral Immune Responses to Those Obtained with Alum.

Biodegradables Chitosan-based Nanoparticles (CS NPs) have been extensively studied as delivery system for therapeutic molecules and as efficient carriers or adjuvants in experimental vaccination. Physicochemical association between CS NPs and antigens is a key step for the biological function as carrier devices. However, for the adjuvant CS NPs property, it is not well known if coupling with vaccine antigens is required or not to potentiate the immune response. To address this issue, in this work, we evaluated the potential adjuvant effect of CS NPs by simply mixing with two different antigens such as Bovine Serum Albumin (BSA) or E protein from Dengue Virus serotype 2 (E protein DENV2). Thus the CS NPs were prepared by ionic gelation with sodium tripolyphosphate, resulting particles among 68 and 188 nm of size. Immunization of 6­8 week old female BALB/c mice, were carried out by intraperitoneal route with a simple combination of CS NPs either with BSA (CS NPs-BSA) at 10 µg or with E protein DENV2 (CS NPs-Protein E) at 5 µg. Combinations with the above antigens with CS NPs elicited robust specific primary and secondary humoral responses comparable to alum, a well-known adjuvant. BSA-specific IgG titers were detectable by day 14 after priming with the CS NPs-BSA formulation, with titers that ranged from 102 to 103 EU ml-. After a second immunization, the anti-BSA titers ranged around 104 EU ml-. In contrast, in the group of mice immunized with the protein alone, BSA-specific serum IgG titers were undetectable at day 14 and 28. For the immunizations with the CS NPs-E protein formulation, we observed also a remarkable specific-antibody production in the primary response, with titers reaching 103 EU ml-. After the booster immunization the anti-E protein DENV2 antibodies titers reached peak values around 104 EU ml-. Interestingly, for both antigens, the combination with CS NPs polarized the immune response to a Th2-like profile, which is characterized mainly by the production of the IgG1 Isotype, confirming that CS NPs can enhance and modulate the humoral immune responses against different antigens independently of physicochemical conjugation. This could represent a simplification in the use of CS NPs as adjuvants in vaccination.

Nodular colitis: endoscopic image an unusual finding.

An 82-year-old male with a history of high blood pressure, COPD, chronic myeloid leukemia, and stage-4 chronic renal failure. Admitted to hospital for lower-limb cellulitis and severe COPD exacerbation, he received antibiotic therapy and bronchodilators. During his hospital stay he developed severe anemia and had an hematochezia event with no diarrhea. A complete colonoscopy found small (4-7 mm) nacreous elevated lesions, circumferential in shape, in the cecum and ascending colon with some bleeding stigmata and submucosal bleeding suggestive of infectious colitis; stool culture was negative and Clostridium difficile toxins were positive. The condition was histologically confirmed.

Utility of artificial intelligence for detection of pneumothorax on chest radiopgraphs done after transthoracic percutaneous transthoracic biopsy guided by computed tomography.

OBJETIVE: To assess the ability of an artificial intelligence software to detect pneumothorax in chest radiographs done after percutaneous transthoracic biopsy. MATERIAL AND METHODS: We included retrospectively in our study adult patients who underwent CT-guided percutaneous transthoracic biopsies from lung, pleural or mediastinal lesions from June 2019 to June 2020, and who had a follow-up chest radiograph after the procedure. These chest radiographs were read to search the presence of pneumothorax independently by an expert thoracic radiologist and a radiodiagnosis resident, whose unified lecture was defined as the gold standard, and the result of each radiograph after interpretation by the artificial intelligence software was documented for posterior comparison with the gold standard. RESULTS: A total of 284 chest radiographs were included in the study and the incidence of pneumothorax was 14.4%. There were no discrepancies between the two readers' interpretation of any of the postbiopsy chest radiographs. The artificial intelligence software was able to detect 41/41 of the present pneumothorax, implying a sensitivity of 100% and a negative predictive value of 100%, with a specificity of 79.4% and a positive predictive value of 45%. The accuracy was 82.4%, indicating that there is a high probability that an individual will be adequately classified by the software. It has also been documented that the presence of Port-a-cath is the cause of 8 of the 50 of false positives by the software. CONCLUSIONS: The software has detected 100% of cases of pneumothorax in the postbiopsy chest radiographs. A potential use of this software could be as a prioritisation tool, allowing radiologists not to read immediately (or even not to read) chest radiographs classified as non-pathological by the software, with the confidence that there are no pathological cases.

Influence of the placebo effect on oxidative stress in healthy older adults of Mexico City.

BACKGROUND: The placebo effect has been widely recognized in the randomized clinical trials; nevertheless, this effect has not been evaluated in terms of antioxidant therapy on oxidative stress (OxS). OBJECTIVE: The objective of this study was to determine the influence of the placebo effect on OxS in healthy older adults of Mexico City. METHODS: We carried out a double-blind controlled clinical assay with the participation of 75 healthy older adults residents for the past 10 years of Mexico City; randomly distributed into three groups of 25 subjects each after previous informed consent; control group not received any treatment, placebo group received a placebo with a pharmaceutical presentation similar to that of the treatment, whereas treatment group were administered 1000 mg of ascorbic acid and 400 IU of alpha-tocopherol. All subjects ingested the treatment daily according to study group for 12 months. We measured before and after 12 months of treatment, lipoperoxides levels (LPO), erythrocyte superoxide dismutase, glutathione peroxidase and total plasma antioxidant status with Randox Laboratories Ltd kits. The concentration of ascorbic acid and alpha-tocopherol were measured using high-performance liquid chromatography. RESULTS: The placebo group subjects showed a statistically significant decrease in LPO concentration, in the same way as the treatment group subjects (P < 0.01), both in comparison with a control group. CONCLUSION: Our findings suggest that the placebo has a significant effect on OxS.

Prevalence of celiac disease and related antibodies in patients diagnosed with irritable bowel syndrome according to the Rome III criteria. A case-control study.

BACKGROUND: The cost-effectiveness for screening for celiac disease (CD) in patients with irritable bowel syndrome (IBS), specifically in the diarrhea (IBS-D) subtype, is beneficial if the prevalence is >1%. However, recent studies have shown controversial results. In this large case-control study, our aim was to determine the prevalence of CD and a panel of related antibodies in patients diagnosed with IBS. MATERIALS AND METHODS: Four hundred IBS patients (Rome III) and 400 asymptomatic healthy controls were prospectively evaluated using antihuman tissue transglutaminase (h-tTG IgA) and deamidated gliadin peptide antibodies (DGP II IgA and DGP II IgG). Duodenal biopsy was performed on the patients that were positive for the h-tTG IgA and/or DGP II IgG antibodies. RESULTS: The mean age of the population was 44.47 ± 18.01 years and 335 (82%) of the subjects were women. Twenty-one patients and six controls had at least one positive test for CD (5.25% VS 1.5%, p = 0.003, OR 3.63 [95% CI 1.4-9.11]). Eighteen patients were positive for h-tTG and/or DGP-II IgG. Histologic confirmation of CD was 2.5% in the IBS patients vs 0.5% in the controls (p = 0.04, OR 5.21). The IBS-D subtype had the highest prevalence for serological positivity (12.7%). CONCLUSIONS: Up to 5.2% of the patients with IBS according to the Rome III criteria were positive for at least one of the CD-related antibodies and 2.5% had biopsy-confirmed CD. Therefore, in our population, screening for CD in subjects with IBS appears to be a reasonable strategy, especially in the IBS-D subgroup.

Inhibition of platelet phosphatidylinositol synthetase by an analog of CDP-diacylglycerol.

Unpublished portions of the synthesis of a phosphinate-phosphonate diether analog of CDPdiacylglycerol are reported. The liponucleotide analog was found to be a very powerful inhibitor of platelet PI synthetase; kinetic data suggest a competitive inhibition mechanism. The structural specificity of CDPdiacylglycerol for liponucleotide-mediated biosynthetic reactions is discussed.

Frontal lobe of children with schizophrenia spectrum disorders: a proton magnetic resonance spectroscopic study.

BACKGROUND: Schizophrenia is commonly considered a neurodevelopmental disorder. Our aim was to determine whether the proton magnetic resonance spectroscopic (1H-MRS) changes seen in adults with schizophrenia are displayed in children at risk for developing schizophrenia. METHODS: Children with symptoms of schizophrenia-spectrum disorders (n = 16; mean age = 132 months) and a comparison group (n = 12; mean age 130 months) took part in a 1H-MRS study of the left frontal lobe. Areas of peaks from N-acetylaspartate (NAA), choline (Cho), and creatine (Cre) were determined and ratios of NAA/Cre and Cho/Cre calculated and compared between groups. RESULTS: The mean ratio of NAA/Cre was significantly lower in schizophrenia-spectrum subjects than the comparison group (1.67 vs. 1.92; p < .05). Medication status did not affect results in schizophrenia-spectrum subjects. CONCLUSIONS: Our findings suggest that the metabolic changes associated with adult schizophrenia are observed in children with some or all of the symptoms of schizophrenia, supporting a neurodevelopmental theory for schizophrenia.

Exploring the multidimensional aspects of grief reactions.

The authors explored the multidimensional aspects of early grief reactions in 201 relatives and close friends of victims of accidental deaths, suicides, homicides, and sudden natural deaths. A principal components analysis of a grief reaction measure revealed four factors: depressive symptoms, preservation of the lost object, suicidal ideation, and decedent-directed anger. The authors suggest that certain features which have been associated mostly with pathological grief (e.g., difficulty relinquishing the lost object) may be present in many, if not most, of the general population during bereavement and that pathology may best be determined by measuring the frequency and intensity of these features.

Magnetic resonance imaging of severely disturbed children--a preliminary study.

This study investigates the relationship between brain pathology and psychiatric disturbance in 37 psychiatric inpatients between 5 and 14 years of age referred for magnetic resonance imaging (MRI). Of 37 images, 13 were categorized as abnormal by neuroradiologists who were blind to the diagnoses of subjects. Three of six children with schizophrenia-related diagnoses had abnormal scans. In contrast, only one of 15 children with a primary disruptive behavior disorder diagnosis had an abnormal magnetic resonance image. A greater proportion of children with schizophrenia spectrum diagnoses had greater left than right frontal horns of the lateral ventricles than children with other diagnoses.

Neuropsychophysiological study of children at risk for schizophrenia: a preliminary report.

OBJECTIVE: This initial report, from an ongoing study, examines whether children who have symptoms of schizophrenia spectrum disorder display neuropsychological or neuroanatomic abnormalities similar to those seen in adults with schizophrenia. METHOD: Experimental subjects were 12 children between 8 and 12 years of age who displayed symptoms of early-onset schizophrenia or schizotypal personality disorder, as assessed through the Schedule for Affective Disorders and Schizophrenia for School-Age Children. The experimental subjects were compared with 13 controls on neuropsychological test performance, magnetic resonance imaging measurements, and proton magnetic resonance spectroscopy results. RESULTS: Findings from the first phase of this project reveal significant overall group differences for several morphometric magnetic resonance imaging measurements and all neuropsychological measures. Differences between the groups were found for amygdala volume, mesial temporal volume, callosal area, and anatomic asymmetry. Magnetic resonance spectroscopy data showed a trend toward group differences. CONCLUSIONS: These findings support a neurodevelopmental model of schizophrenia which postulates that environmentally or genetically programmed events in utero disrupt the establishment of fundamental aspects of brain structure and function.

Synthesis of DL-2,3-diacyloxypropylphosphonyl-cholines from DL-2,3-diacyloxyiodopropanes.

The chemical synthesis of racemic diacyloxypropylphosphonylcholines having octanoyl, myristoyl, oleoyl and stearoyl groups is described. The route involved reaction of dioactanoyloxy-dimyristoyloxy-dioleoyloxy-, and distearoyloxpropyliodide with tris (trimethylsilyl) phosphite to yield the corresponding bis (trimethylsilyl) phosphonate. Removal of the trimethylsilyl groups by neutral aqueous hydrolysis gave the free diacylpropylphosphonic acids, which, when treated with choline toluenesulfonate, yielded the desired dioctanoyloxy-, dimyristoyloxy-, dioleoyloxy-, and distearoyloxypropylphosphonylcholines. The paper also describes the synthesis of 2-octadecyleicosylphosphorylcholine.

Brain abnormalities in schizophrenia-spectrum children: implications for a neurodevelopmental perspective.

Children with symptoms of schizophrenia-spectrum disorder (N = 20) were compared to controls (N = 20) matched for age and socioeconomic status. Structural brain abnormalities were assessed with magnetic resonance imaging and functional brain abnormalities with neuropsychological tests. Children with schizophrenia-spectrum disorder had smaller amygdala and temporal cortex volumes, along with reduced callosal areas and an unusual pattern of neuroanatomic asymmetries. No differences were noted in overall brain volume, ventricular volume, hippocampal volume, or frontal area. Schizophrenia-spectrum children were also characterized by deficits in all neuropsychological functions examined. Some types of verbal memory and frontal lobe skills were especially deficient. These results support the hypothesis that children with schizophrenia-spectrum disorder have significant brain abnormalities, similar in some ways to those seen in adult schizophrenics. In conjunction with recent primate studies, the current results draw attention to the role of the amygdala as one relevant factor in the pathogenesis of schizophrenia.

Dynamics of interleukin-21 production during the clinical course of primary and secondary dengue virus infections.

Previous studies have revealed the clinical relevance of pro-inflammatory cytokine production during dengue virus (DENV) infections. In this study, we evaluated the production of interleukin-21 (IL-21), a key soluble mediator mainly produced by CD4+ T cells. The aim of this study was to investigate the role of IL-21 production during the clinical course of primary and secondary DENV infections and the potential association of IL-21 serum levels with the disease pathogenesis. Blood samples from DENV-infected patients were collected on different days after the onset of symptoms. Patients were classified according to their phase of disease (acute vs. convalescent phases), the type of infection (primary vs. secondary), and the clinical severity of their disease (dengue fever (DF) vs. dengue hemorrhagic fever (DHF)). IL-21 levels were measured using a quantitative capture ELISA assay. The levels of IL-21 were significantly elevated in the disease group compared with the control group. IL-21 was detected in primary and secondary DENV infections, with a significantly higher concentration in the convalescent phase of primary infections. IL-21 levels were significantly higher in patients with secondary acute DHF infections when compared with those with secondary acute DF infection. There was a relationship between the elevated serum levels of IL-21 and the production of DENV-specific IgM and IgG antibodies. Taking together, our results show for the first time the involvement of IL-21 during the clinical course of DENV infections. We speculate that IL-21 may play a protective role in the context of the convalescent phase of primary infections and the acute phase of secondary infections.

Fever, haematuria, and acute graft dysfunction in renal transplant recipients secondary to adenovirus infection: two case reports.

We report two cases of adenoviral infection in kidney transplant recipients that presented with different clinical characteristics under similar demographic and posttransplant conditions. The first case presented with fever, gross haematuria, and acute graft dysfunction 15 days following renal transplantation. A graft biopsy, analyzed with immunohistochemistry, yielded negative results. However, the diagnosis was confirmed with blood and urine real-time PCR for adenovirus 3 days after the initial clinical manifestations. The immunosuppression dose was reduced, and ribavirin treatment was started, for which the patient quickly developed toxicity. Antiviral treatment allowed for transient response; however, a relapse occurred. The viral real-time PCR became negative upon immunosuppression reduction and administration of IVIG; graft function normalized. In the second case, the patient presented with fever and dysuria 1 month after transplantation. The initial imaging studies revealed graft enlargement and areas of hypoperfusion. In this case, the diagnosis was also confirmed with blood and urine real-time PCR for adenovirus 3 days after the initial clinical manifestations. Adenoviral nephritis was confirmed through a graft biopsy analyzed with light microscopy, immunohistochemistry, and PCR in frozen tissue. The immunosuppression dose was reduced, and IVIG was administered obtaining excellent clinical results along with a negative real-time PCR.

Height and weight charts for Mexican adults.

The relation between height and weight is one of the criteria most widely used by doctors to evaluate nutritional condition. The simplest way to establish this relationship is to consult a chart that indicates what a person's weight should be in relation to his height. The charts for adults used in Mexico are deficient, since their design was based on popluations from other countries. In this study the advantages and limitations of this method for evaluating nutritional condition are pointed out and charts designed with data on Mexican university students are presented and compared with the charts that are most frequently being used in Mexico now.

Merolicos and health education.

PIP: Research on merolicos provides valuable information about receptiveness to public health communication. Merolicos are Mexican medicine showmen who by entertaining means such as ventriloquism, mental telepathy, and snakehandling captivate audiences while imparting medicinal information. It was observed that trust in the merolicos extends to people seeking out advice, consultation and explanations. The effective folk traditional health orientation, accessibility and communicative style of the showmen were appealing aspects of the merolicos to the townspeople. In a pilot project the medicine show method of communication was explored in a specific area of infant nutrition in order to evaluate whether the showmen were able to bring about changes in the knowledge, attitudes and behavior patterns of their audience. This involves community selection, message content control, preparation and surveys. Impressive results were seen in adolescents who showed curiosity and attentiveness during the shows. Many young girls valued information about infant care and nutrition. Mexican medicine showmen combine an ability to dramatically work information into community health programs at low potential cost while discreetly disseminating vital health oriented information.^ieng