Characterization of the C6D7-RBD Human Monoclonal Antibody Specific to the SARS-CoV-2 S Protein Receptor-Binding Domain.

The new coronavirus infection COVID-19 is an acute viral disease that affects primarily the upper respiratory tract. The etiological agent of COVID-19 is the SARS-CoV-2 RNA virus (Coronaviridae family, Betacoronavirus genus, Sarbecovirus subgenus). We have developed a high-affinity human monoclonal antibody, called C6D7-RBD, which is specific to the S protein receptor-binding domain (RBD) from the SARS-CoV-2 Wuhan-Hu-1 strain and exhibits virus-neutralizing activity in a test with recombinant antigens: angiotensin-converting enzyme 2 (ACE2) and RBD.

Long-term evaluation of phrenic nerve pacing for respiratory failure due to high cervical spinal cord injury.

OBJECTIVES: Phrenic nerve pacing is a method of respiratory support that can replace mechanical ventilation in high-level cervical spinal cord injury patients with diaphragmatic paralysis. Our objective was to evaluate survival and long-term quality of life in patients with external respiratory support by PNP vs volumetric respirator in patients with severe respiratory insufficiency due to a high-level spinal cord injury. DESIGN: This is a retrospective review study of a prospectively collected database for evaluate the survival and a questionnaire for quality of life has been collected face-to-face or by telephone at present. PATIENTS: Cervical SCI patients with permanent respiratory support (PNP or MV). METHODS: Long-term evaluation of a cohort of PNP-supported patients. We performed a comparison between these patients and volumetric respirator-supported patients. For survival analysis, we used the Kaplan-Meier method and Cox proportional hazards model. The health-related quality of life was assessed with SF-36 questionnaire, a general HRQL evaluation. RESULTS: One hundred twenty six patients on permanent respiratory support were evaluated during the study period. Of these, 38 were on PNP and 88 were mechanically ventilated. Paced patients were younger and had a longer survival, but in a multivariate analysis adjusted for age using a multiple logistic correlation we found that length of survival was greater for PNP patients. In terms of HRQL, the PNP-supported patients showed better results in terms of social functioning. CONCLUSIONS: PNP is a stable and effective method of long-term respiratory support in this type of patients (SCI patients dependent on external respiratory support). In these patients it improves the length of survival and some social issues by quality of life when compared with patients under MV.

Production and Characterization of Rat Monoclonal Antibodies against the PAL Antigen of Legionella spp.

The purpose of this work was to obtain genus-specific monoclonal antibodies against the Legionella spp. recombinant PAL protein, which will subsequently allow to use them as a basis for the development of new express tests for pathogenic legionella detection. A short three-week immunization protocol for Wistar rats was used to generate rat-mouse heterohybridomas producing antibodies against PAL. Mouse myeloma cell line Sp2/0-Ag14 served as the fusion partner. Hybridization was performed using two methods: PEG-mediated fusion and electrofusion. Subsequent screening was performed by indirect solid-phase ELISA against the target protein rPAL. Specificity analysis was performed by dot-blot using a panel of lysates obtained from 39 pure cultures of different strains, which included closely related and heterologous microorganisms among others. No difference in the efficiency of stable hybridoma clones production by the two indicated cell-fusion methods was detected. Twelve clones producing specific rat monoclonal antibodies were obtained based on the screening results. The obtained rat monoclonal antibodies are highly specific towards the PAL protein of L. pneumophila of different serological groups and other pathogenic legionella and are good candidates to be used as the components of diagnostic test systems for the detection of pathogenic representatives of the Legionella genus.

Mechanism of Action of Monoclonal Antibodies That Block the Activity of the Lethal Toxin of Bacillus Anthracis.

Neutralization of the lethal toxin of Bacillus anthracis is an important topic of both fundamental medicine and practical health care, regarding the fight against highly dangerous infections. We have generated a neutralizing monoclonal antibody 1E10 against the lethal toxin of Bacillus anthracis and described the stages of receptor interaction between the protective antigen (PA) and the surface of eukaryotic cells, the formation of PA oligomers, assembly of the lethal toxin (LT), and its translocation by endocytosis into the eukaryotic cell, followed by the formation of a true pore and the release of LT into the cell cytosol. The antibody was shown to act selectively at the stage of interaction between Bacillus anthracis and the eukaryotic cell, and the mechanism of toxin-neutralizing activity of the 1E10 antibody was revealed. The interaction between the 1E10 monoclonal antibody and PA was found to lead to inhibition of the enzymatic activity of the lethal factor (LF), most likely due to a disruption of true pore formation by PA, which blocks the release of LF into the cytosol.

[The participation of the pharmacist in the design and follow-up of the drug treatment plan for patients with a cardiovascular condition]. / Diseño y seguimiento del plan farmacoterapéutico del paciente con enfermedad cardiovascular.

OBJECTIVES: To systemise the pharmaceutical care provided to patients with chronic diseases. To evaluate the pharmacist's participation in the drug treatment plan, studying their intervention in the reconciliation of the patient's habitual treatment and the detection and resolution of drug-related problems. METHOD: A multicentre study based on the comparison of two cohorts: one with the intervention of the pharmacist and one without. Inclusion criteria were as follows: patients over the age of 70 with chronic cardiovascular conditions being treated with more than 6 drugs. They were selected between 24-48 hours from admittance; a control patient was chosen for each patient in the intervention group. The pharmaceutical intervention consisted of medication reconciliation on admittance, drug treatment monitoring and reconciliation on discharge. Drug-related problems, their seriousness, the pharmaceutical intervention, the degree of resolution and the clinical outcomes on discharge were all recorded. A total of 24 hospitals participated, with a total of 356 patients: 180 in the intervention cohort and 176 in the control one. RESULTS: A total of 602 drug-related problems were identified: 66.9% belonging to the intervention group and 33% to the control group. Interventions were made in 359 (89%) patients belonging to the intervention group, 66% were resolved after the pharmaceutical intervention, producing a total or partial improvement in the patient in 36.3% of cases. CONCLUSIONS: Pharmaceutical care has been systematised, providing an instrument that enables all the hospitals to work in a standardised manner. The active participation of the pharmacist in the healthcare team contributes to preventing and resolving drug-related problems.

DNA Barcoding of an Assembly of Montane Andean Butterflies (Satyrinae): Geographical Scale and Identification Performance.

DNA barcoding is a technique used primarily for the documentation and identification of biological diversity based on mitochondrial DNA sequences. Butterflies have received particular attention in DNA barcoding studies, although varied performance may be obtained due to different scales of geographic sampling and speciation processes in various groups. The montane Andean Satyrinae constitutes a challenging study group for taxonomy. The group displays high richness, with more of 550 species, and remarkable morphological similarity among taxa, which renders their identification difficult. In the present study, we evaluated the effectiveness of DNA barcodes in the identification of montane Andean satyrines and the effect of increased geographical scale of sampling on identification performance. Mitochondrial sequences were obtained from 104 specimens of 39 species and 16 genera, collected in a forest remnant in the northwest Andes. DNA barcoding has proved to be a useful tool for the identification of the specimens, with a well-defined gap and producing clusters with unambiguous identifications for all the morphospecies in the study area. The expansion of the geographical scale with published data increased genetic distances within species and reduced those among species, but did not generally reduce the success of specimen identification. Only in Forsterinaria rustica (Butler, 1868), a taxon with high intraspecific variation, the barcode gap was lost and low support for monophyly was obtained. Likewise, expanded sampling resulted in a substantial increase in the intraspecific distance in Morpho sulkowskyi (Kollar, 1850); Panyapedaliodes drymaea (Hewitson, 1858); Lymanopoda obsoleta (Westwood, 1851); and Lymanopoda labda Hewitson, 1861; but for these species, the barcode gap was maintained. These divergent lineages are nonetheless worth a detailed study of external and genitalic morphology variation, as well as ecological features, in order to determine the potential existence of cryptic species. Even including these cases, DNA barcoding performance in specimen identification was 100% successful based on monophyly, an unexpected result in such a taxonomically complicated group.

Graphita gen. nov., a New Genus for Neonympha griphe C. Felder & R. Felder, 1867 (Lepidoptera, Nymphalidae, Satyrinae).

A new genus is described for Neonympha griphe C. Felder & R. Felder, 1867, to contribute toward a revision of the polyphyletic genus Euptychoides Forster, 1964. Based on DNA sequence data, Graphita Nakahara, Marín & Barbosa, gen. nov. is strongly supported as a member of a clade of predominantly southeastern Brazilian taxa, in which it is weakly supported as sister to a well-supported clade containing Pharneuptychia Forster, 1964, Moneuptychia Forster, 1964 and the E. castrensis (Schaus, 1902) species complex. The data show that Graphita griphe comb. nov. is not related to other Euptychoides and not very closely related to any other sampled euptychiines, and thus support the description of this new genus. In addition, we provide morphological illustrations and a distribution map for this taxon based on museum specimens.

[Neuropsychiatric disorders associated with streptococci: a case report]. / Trastornos neuropsiquiátricos asociados a estreptococo.

Compulsive neuropsychiatric disorders are common in children and the most frequent is Gilles de la Tourette syndrome. Recently, a new disease has been described: the PANDAS syndrome (pediatric autoimmune neuropsychiatric disorders associated with streptococci). The etiology of this syndrome is uncertain but it has been associated with recent group A streptococcal infection (GAS). After an episode of pharyngitis, a boy aged 3 years and 9 months showed abrupt onset of a variety of neurobehavioral problems such as tics (consisting of elevation of the head and ipsilateral shoulder, winking, and grimaces) and compulsions (such as repeatedly hitting objects). A complete biochemical study was performed, including Cu and ceruloplasmin (which had normal values), antistreptolysin O (ASLO) and anti-DNAse (showing elevated values). Pharyngeal culture revealed GAS. The child was initially treated with valproic acid, but his subsequent improvement coincided with penicillin treatment for 10 days. Three months afterwards, ASLO values were reduced and at the 6-month follow-up the improvement was maintained even after suspension of valproic acid. Immunohistochemical studies were requested. PANDAS syndrome was first described in 1998 in a group of children who presented 1) obsessive compulsive disorders and/or tics, 2) episodic course with abrupt exacerbations, 3) abnormal results of neurologic examination (choreiform movements), and 4) temporal relation between GAS infection and onset of symptoms. The etiology of this syndrome is unclear, and it has been postulated that certain streptococcal antigens trigger antibodies which, through a process of molecular mimicry, cross-react with epitopes on the basal ganglia of susceptible hosts, such as the B8/17 antigen, among others. Current recommendations include penicillin treatment of each exacerbation with positive throat culture, and more aggressive therapies (intravenous immunoglobulin or plasmapheresis) when symptoms are severe. We believe that the case presented herein is a probable PANDAS syndrome, which would be the first case described in Spain.

Interaction of C3 nephritic factor (NEF) with erythrocyte membranes complement-independent binding to sheep and patients' erythrocytes.

Complement-independent binding of C3 nephritic factor (NEF) to sheep erythrocytes was observed in heat-inactivated sera from patients having this autoantibody. The binding was observed after neuraminidase treatment of erythrocytes but not following trypsin treatment. Purified IgG from patients' sera was able to bind to ShE membranes. Binding to rat and rabbit erythrocytes was also observed but not to human group O+ erythrocytes. By Western blot NEF ab recognizes a 26 kD protein on the sheep erythrocytes and a 21 kD protein on human erythrocytes. NEF activity decreased at these positions when blotted nitrocellulose was incubated with NEF antibody. This autoantibody binds human erythrocytes membranes from patients but not from 55 normal blood donors. IgG from a pool from 10 different controls did not bind membrane E from the patients. The amino acid analysis of the 21 kD protein of the patients showed differences in basic residues (Arg and Lys) when compared with the 21 kD protein obtained from controls. N-terminal sequence analysis indicated that it is blocked in both proteins.

[Perceptions and experiences of access to health services and their utilization among the immigrant population]. / Percepción y experiencias en el acceso y el uso de los servicios sanitarios en población inmigrante.

OBJECTIVE: To identify and describe the needs and problems of the immigrant population related to access and utilization of health services. METHOD: A descriptive, qualitative, phenomenological study was conducted using focus groups. The study area was the county of Campo de Gibraltar (Spain), which represents the gateway to Europe for immigration from Africa. The final sample size (51 immigrants from 11 countries) was determined by theoretical saturation. A narrative analysis was conducted with QSR NVivo9 software. RESULTS: Immigrants' discourse showed four categories of analysis: response to a health problem, system access, knowledge of social and health resources, and health literacy needs. Responses to health problems and the route of access to the health care system differed according to some sociodemographic characteristics (nationality/culture of origin, length of residence, and economic status). In general, immigrants primarily used emergency services, hampering health promotion and prevention. The health literacy needs identified concerned language proficiency and the functioning of the health system. CONCLUSIONS: There is a need to promote interventions to enhance health literacy among immigrants. These interventions should take into account diversity and length of residence, and should be based on an action-participation methodology.

C3 nephritic factor determination. A comparison between two methods.

C3 nephritic factor (NEF), an IgG autoantibody to the alternative pathway C3 convertase, is usually measured by crossed immunoelectrophoresis (CI) but recently a reliable haemolytic assay (HA) was described by Rother (1982). This method is more specific than CI because it is negative in sera with immune complexes, SLE and sera incubated with IgG aggregates. The haemolytic assay is sensitive enough to detect NEF antibody in serum from patients with only slightly low C3 levels and NEF negatives by CI. The haemolytic assay is easy to perform and reproducible, the interassay coefficient of variation being 10.7% compared to 64% in the CI. The intra-assay coefficient of variation in CI was 28% compared to 5.5% in the haemolytic assay. The haemolytic method enabled us to study the kinetic effects of NEF on C3b.Bb bound to sheep erythrocytes, and the lysis mediated by ShE.C3b.Bb.NEF complex. Also the C and NEF binding to sheep erythrocytes was studied.

Human leukocyte antigen-associated susceptibility to pulmonary tuberculosis: molecular analysis of class II alleles by DNA amplification and oligonucleotide hybridization in Mexican patients.

BACKGROUND: Pulmonary tuberculosis (PTB) develops by a complex combination of environmental factors with genetic susceptibility. In this context, an association between human leukocyte antigens (HLAs) and tuberculosis has been examined in several populations, but results have been controversial. DESIGN AND MEASUREMENTS: A prospective evaluation of class II HLA genotypes was completed by the polymerase chain reaction (PCR) sequence-specific primer technique and PCR sequence-specific oligonucleotide hybridization in a Mexican population. SETTING: This study was conducted at the Clinical Service of Tuberculosis and the Department of Immunology, National Institute of Respiratory Diseases, Mexico City, Mexico. PATIENTS: Four groups were examined: 95 healthy subjects; 50 nonimmunosuppressed PTB patients; 15 HIV-infected patients (stage IVc in the Centers for Disease Control and Prevention [CDC] classification system for AIDS) with PTB; and 37 HIV-infected patients in the asymptomatic stage (CDC stage II). RESULTS: The frequencies of alleles DQA1*0101 (odds ratio [OR], 6.18; 95% confidence interval [CI], 2.38 to 16.08), DQB1*0501 (OR, 6.16; 95% CI, 2.44 to 17.71), and DRB1*1501 (OR, 7.92; 95% CI, 2.71 to 23.14) were significantly increased in nonimmunosuppressed patients with PTB when compared with healthy subjects. By contrast, frequencies of allele DQB1*0402 and antigens DR4 and DR8 were significantly decreased in patients with PTB. Additionally, a significantly higher frequency of the DRB1*1101 allele was found in HIV-positive subjects (OR, 6.67; 95% CI, 2.13 to 20.83). CONCLUSION: The genetic influence associated with the HLA system appears to have an important role in the development of PTB, although this susceptibility may not be relevant in patients with severe immunodeficiency diseases such as AIDS.

Dengue epidemics on the Pacific Coast of Mexico.

Dengue epidemics in three Mexican cities were investigated with retrospective surveys and virus isolations from acute cases. These epidemics were part of the continuing extension of dengue in Mexico since 1978 after 15 years without transmission. Serotype 1 dengue infection predominated in all epidemics, but in one city, type 2 strains were also isolated. The following findings were consistent in all three cities: 1) illness history provided evidence of presence of infection months before the epidemic became evident, 2) there was a very sharp and progressive increase of dengue illness attack rates by age, 3) there were no interpretable relationships between illness and the presence of different types of breeding sights in the home, 4) socioeconomic status was strongly negatively related to illness, and 5) geographical factors not related to the other measured variables had a strong effect on illness rates reflecting the focality of transmission. In addition, there was evidence of a strong protective effect against illness by the use of mosquito netting or the presence of screens on houses, but these relationships were not universal to all three cities.

Determination of C3 nephritic factor activity by a microassay based on the peroxidase-like activity of the heme group.

We report a modification of a method for measuring the C3 nephritic factor, using the peroxidase activity of the heme group. This modification increases the sensitivity of the method, approximately seven-fold when NEF activities are measured in U/mL. It is less time-consuming and it allows the simultaneous testing of a larger number of samples, simplifying the screening of sera and ensuring an easy and simple test for the detection of NEF in purification processes as well as in in vitro production. The method could also be useful in other hemolytic assays.

Anopheles vestitipennis, the probable vector of Plasmodium vivax in the Lacandon forest of Chiapas, México.

Baseline studies to evaluate the role of different anopheline mosquitoes as probable vectors of malaria in the Lacandon rainforest region in Chiapas (México) were carried out in 3 rural villages between June and November 1988. Anopheles vestitipennis was the most abundant species in all the villages, followed by An. albimanus, An. punctimacula and An. darlingi. Human bait collections showed that An. vestitipennis and An. albimanus tended to bite man either indoors or outdoors, while An. punctimacula preferred to bite outdoors. Monthly man-biting rates of An. vestitipennis tended to follow the rainfall pattern. Plasmodium vivax was detected in An. vestitipennis only, with an estimated overall infection rate of 4.67 per 1000 mosquitoes (range 0-54.42, of 3500 mosquitoes assayed). This is the first incrimination of An. vestitipennis as a probable vector of P. vivax in México.

Clinical application of amniotic membranes on a patient with epidermolysis bullosa.

The case of a patient with dystrophic epidermolysis bullosa treated with radiosterilised amniotic membranes is presented. The disorder is a congenital disease characterised by a poor desmosomal junction in the keratinocyte membrane. After proper donor screening, amnios were collected at Hospital Central Sur de Alta Especialidad (HCSAE), PEMEX and microbiological analysis was performed at Universidad Nacional Autónoma de México, FQUNAM, (Biology Dept. of the Chemistry Faculty, National Autonomous University of Mexico), before and after radiation sterilisation. Processing, packaging and sterilisation were performed at Instituto Nacional de Investigaciones Nucleares, ININ, (National Nuclear Research Institute). The patient, a ten-year-old boy with severe malnutrition, extensive loss of skin and pseudomonad infection in the whole body, was treated with gentle debridement in a Hubbard bath. Later amnion application was performed with sterilised amnios by using two different processes, in one of which the amnion was sterilised with paracetic acid, preserved in glycerol, kindly donated by the German Institute for Tissue and Cell Replacement and applied by Dr. Johannes C. Bruck, IAEA visiting expert, and the other amnion was processed at ININ: air dried and sterilised by gamma radiation at dose of 30 kGy. After spontaneous epithelisation was successfully promoted for seven days, the pain was alleviated and mobility was improved in a few hours and the patient's general condition was so improved that in a month he was discharged. Unfortunately, because this disease is revertive and has malignant degeneration, the prognosis is not good.