Molecular defects of the C7 gene in two patients with complement C7 deficiency

Different genetic mutations have been described in complement components resulting in total or subtotal deficiency states. In this work we report the genetic basis of C7 deficiency in a previously reported Spanish patient exhibiting a combined total deficiency of C7 and C4B associated with systemic lupus erythematosus. Exon-specific polymerase chain reaction and sequencing revealed a not previously described single base mutation in exon 10 (T1458A) leading to a stop codon that causes the premature truncation of the C7 protein (C464X). Additionally, a C to A transversion at position 1561 (exon 11) was found in the patient resulting in an amino acid change (R499S). This latter mutation has been previously reported in individuals with subtotal C7 deficiency or with combined subtotal C6/C7 deficiency from widely spaced geographical areas. Another novel mutation was found in a second patient with meningococcal meningitis of Bolivian and Czech origin; a 11-base pair deletion of nucleotides 631-641 in exon 6 leading to the generation of a downstream stop codon causing the premature truncation of the C7 protein product (T189 x 193). This patient was found to be a heterozygous compound for another mutation in C7; a two-base pair deletion of nucleotides 1922 and 1923, 1923 and 1924 or 1924 and 1925 in exon 14 (1922delAG/1923delGA/1924delAG), leading again to the generation of a downstream stop codon that provokes the truncation of the C7 protein (S620x630). This latter mutation has been recently reported by our group in another Spanish family. Our results provide more evidences for the heterogeneous molecular basis of C7 deficiency.     

IgE antibodies to betalactams: relationship between the triggering hapten and the specificity of the immune response

BACKGROUND: In immunoglobulin (Ig)E-mediated responses to betalactams (BL) the antibody is directed to the hapten inducing the response. For benzylpenicillin (BP) the determinant is benzylpenicilloyl (BPO) and for amoxicillin (AX), amoxicilloyl (AXO). Because of cross-reactivity, IgE from some patients reacts to both drugs whereas others have a drug-selective recognition. After an allergic episode, there is an increase in IgE that decreases over time. We analysed the response of patients allergic to BL after penicillin administration, with emphasis on IgE cross-reactivity. METHODS: Subjects who developed an IgE antibody response were studied. Sequential follow-up samples were obtained at different times during the response. Changes in IgE specificity were analysed by competition immunoassays using different penicillin monomeric conjugates. RESULTS: Two patterns of response were existed: one with IgE directed to the culprit penicillin and another with IgE mainly reactive to BPO. In both, a variable cross-reactivity with the hapten triggering the boosting response was found. This pattern was maintained with no change in specificity over time, even in subjects who experienced one boosting event. CONCLUSION: The IgE response can be specific to the drug inducing the reaction or cross-reactive to the classical BPO determinant. This pattern is maintained throughout the whole period of the response, even if re-exposure occurs. The stability of the type of response can be explained by the phenomenon of original antigenic sin: in the presence of antibodies, memory B cells are more easily triggered than naive B cells.     

Syncytial giant cell hepatitis in human immunodeficiency virus-infected patients with chronic hepatitis C: 2 cases and review of the literature

Syncytial giant cell hepatitis (SGCH) among adult human immunodeficiency virus (HIV)-infected patients has been rarely described. Most cases have been reported in subjects coinfected with the hepatitis C virus (HCV), but its prevalence and outcome remain unknown. We performed a retrospective analysis of all cases of SGCH among 332 liver biopsies from HIV-infected patients seen at a tertiary center in Madrid, Spain, between 1984 and March 2004. Two hundred fifty specimens were obtained from HCV-coinfected patients. There were 2 cases of SGCH, leading to an observed overall prevalence of 0.6% (0.8% when considering only HCV-coinfected patients). In addition to histological changes secondary to chronic hepatitis C, the liver cords were replaced by syncytial giant cells with up to 30 nuclei. There was no histological evidence of measles (among paramyxoviruses) or herpes viruses group infections. In patient 1, there was a progressive clinical worsening after a 3-month course of prednisone, leading to liver failure and death. His postmortem liver biopsy showed more abundant giant hepatocytes accompanied with the development of a histologic pattern of severe fibrosing cholestatic hepatitis. The second patient received a prolonged course of pegylated interferon-alpha-2b and ribavirin with clearance of syncytial giant hepatocytes despite HCV-RNA persistence. SGCH is a rare histological finding among HIV-infected patients with chronic hepatitis C. Specific treatment with pegylated interferon and ribavirin can lead to histological resolution and biochemical improvement, even in the absence of HCV-RNA clearance.     

Evaluation and genotyping of multidrug-resistant cases of tuberculosis in southern Brazil

Sixty isolates of Mycobacterium tuberculosis identified as multidrug-resistant (MDR) at a reference laboratory in Rio Grande do Sul State during the years 1999 and 2000 were analyzed using the IS6110-restriction fragment length polymorphism (RFLP) technique. We also genotyped 202 susceptible strains to compare the genotyping results, as well as the clinical and demographic data. Spacer oligotyping (spoligotyping) analysis was performed for isolates presenting low IS6110 copy number. Patients with identical DNA pattern strains were considered clustered. From 262 isolates, 94 (36%) belonged to 20 distinct RFLP clusters, and after spoligotyping analysis, 89 of the isolates (34%) remained in cluster. MDR isolates did not differ statistically in clustering proportion from susceptible strains. A significant association between the occurrence of MDR and previous tuberculosis (TB) treatment was observed (p < 0.001), as well as failure on TB treatment (p < 0.001). Human immunodeficiency virus (HIV)-positive patients were associated with susceptible tuberculosis (p = 0.024). We also identified that unmarried patients were more likely to develop TB due to recent transmission than married patients (p < 0.005). The introduction of directly observed therapy short-course (DOTS) strategy will be important in decreasing default and failure rates and avoiding the development of new MDR strains.     

Long-term outcomes of concomitant cisplatin plus radiotherapy versus radiotherapy alone in patients with stage IIIB squamous cervical cancer: A randomized controlled trial

ObjectiveThe present analysis determined the disease free survival (DFS) and overall survival (OS) at up to 14 years of follow-up in women who participated in our previous phase 3 randomized controlled clinical trial, in which women with stage IIIB squamous cervical cancer received either cisplatin plus RT or RT alone for treatment. The first study showed that the addition of cisplatin to RT offered a significant benefit in DFS, but not in OS.MethodsThe present analysis examined DFS and OS in 146 women from the original cohort (72 patients in the CRT arm and 74 patients in the RT-only arm) with follow-up of up to 14 years.ResultsLonger term follow-up showed that treatment with CRT offers a significant benefit in DFS and OS compared with treatment with RT only. Patients who received RT alone had significantly worse OS (HR, 1.88; 95% CI, 1.09–3.24) and DFS (HR, 1.82; 95% CI, 1.07–3.08) compared with patients who received CRT. The multivariate analyses also showed that the patients with baseline Karnofsky performance status (KPS) <90% showed significantly worse OS (HR, 3.11; 95% CI, 1.78–5.43), as did those with hemoglobin <10 mg/dL (HR, 4.32; 95% CI, 2.23–8.36). Patients with baseline KPS < 90% showed significantly worse DFS (HR, 2.83; 95% CI, 1.60–5.01), as did those with hemoglobin <10 mg/dL (HR, 4.16; 95% CI, 2.17–7.95).ConclusionsFor stage IIIB cervical cancer, treatment with CRT offers a significant benefit in DFS and OS compared with treatment with RT only.     

Calidad de vida,complicaciones asociadas y satisfacción con el uso de pesarios para tratamiento conservador del prolapso de órganos pélvicos

BackgroundConservative treatment for pelvic organ prolapse (POP) is indicated when the patient refuses surgery, or when there is surgical contraindication due to adverse medical conditions.ObjectiveTo evaluate the quality of life, complications, and perception of health improvement in women with POP on treatment with pessaries in the Urogynaecology Unit of the Military Hospital, Santiago Chile between 2009 and 2018.MethodRetrospective study that evaluated 60 women with POP on treatment with pessaries. Sociodemographic variables and clinical data were collected at the beginning of the treatment and during follow-up from April 2018 to March 2019. Two validated questionnaires were also completed: Subjective Perception of Improvement and Prolapse Quality of Life (PQoL).ResultsThe ring was the most commonly used pessary (75%). The mean age of the patients was 78.7 years (± 8.2). The most reported complications were: vulvovaginitis, erosion, urinary tract infection, and flange. Around 80% of the patients reported an excellent improvement in their health condition, and 20% reported feeling better. Women with erosion and vulvovaginitis had a longer time of pessary use compared to women who did not have these complications (p < 0.05). In general, Quality of Life associated with prolapse and pessary use was good. Women with urinary incontinence showed a worse quality of life in domains «impact of prolapse» and «social limitations» (p < 0.05).ConclusionConservative treatment with pessaries showed good results in subjective perception of improvement, good quality of life, and low percentage of complications.     

Chronic benzylamine administration in the drinking water improves glucose tolerance,reduces body weight gain and circulating cholesterol in high-fat diet-fed mice

Benzylamine is found in Moringa oleifera, a plant used to treat diabetes in traditional medicine. In mammals, benzylamine is metabolized by semicarbazide-sensitive amine oxidase (SSAO) to benzaldehyde and hydrogen peroxide. This latter product has insulin-mimicking action, and is involved in the effects of benzylamine on human adipocytes: stimulation of glucose transport and inhibition of lipolysis. This study examined whether chronic, oral administration of benzylamine could improve glucose tolerance and the circulating lipid profile without increasing oxidative stress in overweight and pre-diabetic mice. The benzylamine diffusion across the intestine was verified using everted gut sacs. Then, glucose handling and metabolic markers were measured in mice rendered insulin-resistant when fed a high-fat diet (HFD) and receiving or not benzylamine in their drinking water (3600 μmol/(kg day)) for 17 weeks. HFD-benzylamine mice showed lower body weight gain, fasting blood glucose, total plasma cholesterol and hyperglycaemic response to glucose load when compared to HFD control. In adipocytes, insulin-induced activation of glucose transport and inhibition of lipolysis remained unchanged. In aorta, benzylamine treatment partially restored the nitrite levels that were reduced by HFD. In liver, lipid peroxidation markers were reduced. Resistin and uric acid, surrogate plasma markers of metabolic syndrome, were decreased. In spite of the putative deleterious nature of the hydrogen peroxide generated during amine oxidation, and in agreement with its in vitro insulin-like actions found on adipocytes, the SSAO-substrate benzylamine could be considered as a potential oral agent to treat metabolic syndrome.     

Overview and update on molecular testing in non-small cell lung carcinoma utilizing endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) samples

Lung carcinoma remains one of the most frequent and aggressive human neoplasms. Fortunately, in the last decades, the increasing knowledge of the molecular mechanisms leading to cancer development has allowed the use of targeted therapies with improvement of prognosis in many patients. Clinical management has also changed after the introduction of endobronchialultrasonographic bronchoscopy that allows a conservative staging of lung tumors, avoiding the need of mediastinoscopy for lymph node staging. Lung pathologists and cytopathologists are facing the challenge of giving the more comprehensive prognostic and predictive information with ever smaller tissue or cytological samples. The aim of this review is to summarize the molecular testing for non-small cell lung carcinoma and how pathologists can contribute to the patient's outcome with a conscious management of biological samples.