Electrochemical Detection of Total Antioxidant Capacity (TAC) in Plant Tissues from Different Origins.

Total antioxidant capacity (TAC) is a reliable indicator of antioxidant content in animal and plant samples. The different experimental approaches available allow the determination of TAC using, as a reference, diverse compounds with recognized antioxidant capacities such as Trolox, ascorbic acid, gallic acid, or melatonin. A new portable device, named BRS (BQC redox system), is now commercially available that, through an electrochemical approach, allows the determination of TAC in a simple, fast, reproducible, and robust way. In this chapter, using this portable device, a comparative analysis of the TAC is assayed in different red, citrus, and Solanaceae fruits, several Allium species, and organs of different plant species, including Arabidopsis thaliana. The obtained results demonstrate the versatility of the BRS portable device.

Characterization of leucine aminopeptidase (LAP) activity in sweet pepper fruits during ripening and its inhibition by nitration and reducing events.

KEY MESSAGE: Pepper fruits contain two leucine aminopeptidase (LAP) genes which are differentially modulated during ripening and by nitric oxide. The LAP activity increases during ripening but is negatively modulated by nitration. Leucine aminopeptidase (LAP) is an essential metalloenzyme that cleaves N-terminal leucine residues from proteins but also metabolizes dipeptides and tripeptides. LAPs play a fundamental role in cell protein turnover and participate in physiological processes such as defense mechanisms against biotic and abiotic stresses, but little is known about their involvement in fruit physiology. This study aims to identify and characterize genes encoding LAP and evaluate their role during the ripening of pepper (Capsicum annuum L.) fruits and under a nitric oxide (NO)-enriched environment. Using a data-mining approach of the pepper plant genome and fruit transcriptome (RNA-seq), two LAP genes, designated CaLAP1 and CaLAP2, were identified. The time course expression analysis of these genes during different fruit ripening stages showed that whereas CaLAP1 decreased, CaLAP2 was upregulated. However, under an exogenous NO treatment of fruits, both genes were downregulated. On the contrary, it was shown that during fruit ripening LAP activity increased by 81%. An in vitro assay of the LAP activity in the presence of different modulating compounds including peroxynitrite (ONOO-), NO donors (S-nitrosoglutathione and nitrosocyteine), reducing agents such as reduced glutathione (GSH), L-cysteine (L-Cys), and cyanide triggered a differential response. Thus, peroxynitrite and reducing compounds provoked around 50% inhibition of the LAP activity in green immature fruits, whereas cyanide upregulated it 1.5 folds. To our knowledge, this is the first characterization of LAP in pepper fruits as well as of its regulation by diverse modulating compounds. Based on the capacity of LAP to metabolize dipeptides and tripeptides, it could be hypothesized that the LAP might be involved in the GSH recycling during the ripening process.

In Silico RNAseq and Biochemical Analyses of Glucose-6-Phosphate Dehydrogenase (G6PDH) from Sweet Pepper Fruits: Involvement of Nitric Oxide (NO) in Ripening and Modulation.

Pepper (Capsicum annuum L.) fruit is a horticultural product consumed worldwide which has great nutritional and economic relevance. Besides the phenotypical changes that pepper fruit undergo during ripening, there are many associated modifications at transcriptomic, proteomic, biochemical, and metabolic levels. Nitric oxide (NO) is a recognized signal molecule that can exert regulatory functions in diverse plant processes including fruit ripening, but the relevance of NADPH as a fingerprinting of the crop physiology including ripening has also been proposed. Glucose-6-phosphate dehydrogenase (G6PDH) is the first and rate-limiting enzyme of the oxidative phase of the pentose phosphate pathway (oxiPPP) with the capacity to generate NADPH. Thus far, the available information on G6PDH and other NADPH-generating enzymatic systems in pepper plants, and their expression during the ripening of sweet pepper fruit, is very scarce. Therefore, an analysis at the transcriptomic, molecular and functional levels of the G6PDH system has been accomplished in this work for the first time. Based on a data-mining approach to the pepper genome and fruit transcriptome (RNA-seq), four G6PDH genes were identified in pepper plants and designated CaG6PDH1 to CaG6PDH4, with all of them also being expressed in fruits. While CaG6PDH1 encodes a cytosolic isozyme, the other genes code for plastid isozymes. The time-course expression analysis of these CaG6PDH genes during different fruit ripening stages, including green immature (G), breaking point (BP), and red ripe (R), showed that they were differentially modulated. Thus, while CaG6PDH2 and CaG6PDH4 were upregulated at ripening, CaG6PDH1 was downregulated, and CaG6PDH3 was slightly affected. Exogenous treatment of fruits with NO gas triggered the downregulation of CaG6PDH2, whereas the other genes were positively regulated. In-gel analysis using non-denaturing PAGE of a 50-75% ammonium-sulfate-enriched protein fraction from pepper fruits allowed for identifying two isozymes designated CaG6PDH I and CaG6PDH II, according to their electrophoretic mobility. In order to test the potential modulation of such pepper G6PDH isozymes, in vitro analyses of green pepper fruit samples in the presence of different compounds including NO donors (S-nitrosoglutathione and nitrosocysteine), peroxynitrite (ONOO-), a hydrogen sulfide (H2S) donor (NaHS, sodium hydrosulfide), and reducing agents such as reduced glutathione (GSH) and L-cysteine (L-Cys) were assayed. While peroxynitrite and the reducing compounds provoked a partial inhibition of one or both isoenzymes, NaHS exerted 100% inhibition of the two CaG6PDHs. Taken together these data provide the first data on the modulation of CaG6PDHs at gene and activity levels which occur in pepper fruit during ripening and after NO post-harvest treatment. As a consequence, this phenomenon may influence the NADPH availability for the redox homeostasis of the fruit and balance its active nitro-oxidative metabolism throughout the ripening process.

NADP-Dependent Malic Enzyme Genes in Sweet Pepper Fruits: Involvement in Ripening and Modulation by Nitric Oxide (NO).

NADPH is an indispensable cofactor in a wide range of physiological processes that is generated by a family of NADPH dehydrogenases, of which the NADP-dependent malic enzyme (NADP-ME) is a member. Pepper (Capsicum annuum L.) fruit is a horticultural product consumed worldwide that has great nutritional and economic relevance. Besides the phenotypical changes that pepper fruit undergoes during ripening, there are many associated modifications at transcriptomic, proteome, biochemical and metabolic levels. Nitric oxide (NO) is a recognized signal molecule with regulatory functions in diverse plant processes. To our knowledge, there is very scarce information about the number of genes encoding for NADP-ME in pepper plants and their expression during the ripening of sweet pepper fruit. Using a data mining approach to evaluate the pepper plant genome and fruit transcriptome (RNA-seq), five NADP-ME genes were identified, and four of them, namely CaNADP-ME2 to CaNADP-ME5, were expressed in fruit. The time course expression analysis of these genes during different fruit ripening stages, including green immature (G), breaking point (BP) and red ripe (R), showed that they were differentially modulated. Thus, while CaNADP-ME3 and CaNADP-ME5 were upregulated, CaNADP-ME2 and CaNADP-ME4 were downregulated. Exogenous NO treatment of fruit triggered the downregulation of CaNADP-ME4. We obtained a 50-75% ammonium-sulfate-enriched protein fraction containing CaNADP-ME enzyme activity, and this was assayed via non-denaturing polyacrylamide gel electrophoresis (PAGE). The results allow us to identify four isozymes designated from CaNADP-ME I to CaNADP-ME IV. Taken together, the data provide new pieces of information on the CaNADP-ME system with the identification of five CaNADP-ME genes and how the four genes expressed in pepper fruits are modulated during ripening and exogenous NO gas treatment.

Increased Surveillance or Increased Scrutiny: Curbing Inappropriate Screening of Endometrial Cancer in Transgender Men.

Objective: The transgender male body is often considered a "black box" by even the most senior of attending physicians. Because of the stigma and fear associated with the utilization of Hormone Replacement Therapy (HRT) and the lack of clinical knowledge surrounding the subject, many transgender men will experience unnecessary testing for endometrial cancers. Materials and methods: This narrative review looked through publicly available PubMed and GoogleScholar articles on the topic of the use of pelvic ultrasounds in gynecological screening in transgendered males. 18,000 articles were filtered by relevance, date, clarity of the topic, and clinical recommendations. Of these articles 37 were included for discussion. Results: Of these 18,000 articles, only 37 were included for discussion. The resulting table, diagnostic tree, and discussion section are included within. Conclusion: Herein, the authors discuss the current understanding of the role of imaging in the diagnosis and treatment of gynecological cancers in transgendered men and how unnecessary imaging studies can be curbed for the benefit of the patient and the medical system at large.

Diagnostic accuracy of adenosine deaminase for pleural tuberculosis in a low prevalence setting: A machine learning approach within a 7-year prospective multi-center study.

OBJECTIVE: To analyze the performance of adenosine deaminase in pleural fluid combined with other parameters routinely measured in clinical practice and assisted by machine learning algorithms for the diagnosis of pleural tuberculosis in a low prevalence setting, and secondly, to identify effusions that are non-tuberculous and most likely malignant. PATIENTS AND METHODS: We prospectively analyzed 230 consecutive patients diagnosed with lymphocytic exudative pleural effusion from March 2013 to June 2020. Diagnosis according to the composite reference standard was achieved in all cases. Pre-test probability of pleural tuberculosis was 3.8% throughout the study period. Parameters included were: levels of adenosine deaminase, pH, glucose, proteins, and lactate dehydrogenase, red and white cell counts and lymphocyte percentage in pleural fluid, as well as age. We tested six different machine learning-based classifiers to categorize the patients. Two different classifications were performed: a) tuberculous/non-tuberculous and b) tuberculous/malignant/other. RESULTS: Out of a total of 230 patients with pleural effusion included in the study, 124 were diagnosed with malignant effusion and 44 with pleural tuberculosis, while 62 were given other diagnoses. In the tuberculous/non-tuberculous classification, and taking into account the validation predictions, the support vector machine yielded the best result: an AUC of 0.98, accuracy of 97%, sensitivity of 91%, and specificity of 98%, whilst in the tuberculous/malignant/other classification, this type of classifier yielded an overall accuracy of 80%. With this three-class classifier, the same sensitivity and specificity was achieved in the tuberculous/other classification, but it also allowed the correct classification of 90% of malignant cases. CONCLUSION: The level of adenosine deaminase in pleural fluid together with cell count, other routine biochemical parameters and age, combined with a machine-learning approach, is suitable for the diagnosis of pleural tuberculosis in a low prevalence scenario. Secondly, non-tuberculous effusions that are suspected to be malignant may also be identified with adequate accuracy.

Ocular Ischemic Syndrome With Congenital Absence of Internal Carotid Artery.

Purpose: This article describes a case of ocular ischemic syndrome (OIS) in a patient with a congenitally absent left internal carotid artery (ICA). Methods: Retrospective case report with anterior-segment and fundus photography, fluorescein angiography (FA), and computerized tomography angiography (CT-A). Results: A 31-year-old-man was found to have neovascularization of the iris (NVI) and angle of the left eye. FA showed capillary nonperfusion in the temporal periphery. He required intravitreal bevacizumab and triamcinolone injections and 2 panretinal photocoagulation treatments for persistent rubeosis and cystoid macular edema. Transient right-eye vision loss prompted CT-A, revealing an absent left ICA. Three years following presentation, FA continued to show delayed arteriovenous flow suggestive of OIS. He has required intravitreal bevacizumab injections every 12 weeks for persistent NVI. Conclusion: Congenitally absent left ICA with resultant cerebrovascular insufficiency is a rare cause of OIS, underscoring the pathophysiological principles of insufficient blood supply to the ophthalmic artery.

Recurrent epiploic appendagitis mimicking appendicitis and cholecystitis.

Epiploic appendagitis (EA) is a rare cause of acute abdominal pain caused by inflammation of an epiploic appendage. It has a nonspecific clinical presentation that may mimic other acute abdominal pathologies on physical exam, such as appendicitis, diverticulitis, or cholecystitis. However, EA is usually benign and self-limiting and can be treated conservatively. We present the case of a patient with two episodes of EA, the first mimicking acute appendicitis and the second mimicking acute cholecystitis. Although recurrence of EA is rare, it should be part of the differential diagnosis of acute, localized abdominal pain. A correct diagnosis of EA will prevent unnecessary hospitalization, antibiotic use, and surgical procedures.

Valor diagnóstico del ada para tuberculosis pleural en diferentes escenarios de prevalencia de enfermedad / No disponible

La tuberculosis pleural es una manifestación común extrapulmonar de la enfermedad. Su diagnóstico de certeza se basa en la demostración del bacilo o en el hallazgo degranulomas en la biopsia pleural. En las últimas décadas se han desarrollado parámetros alternativos que pueden establecer el diagnóstico con certeza razonable, destacando la Adenosina Desaminasa (ADA) en líquido pleural por su excelente coste-efectividad. En teoría, el valor diagnóstico del ADA se ve disminuido en escenarios de baja prevalencia de enfermedad. Por otro lado la combinación del ADA con el ratio linfocitos/neutrófilos aumenta su especificidad. Analizamos de forma retrospectiva en nuestra comarca el valor de ambas variables combinadas en el diagnóstico de la tuberculosis pleural entres escenarios distintos de prevalencia de TBC pleural en el periodo 1996-2009 (AU)

The many faces of fat necrosis in the breast.

OBJECTIVE: This article describes the manifestations of fat necrosis on mammography, sonography, and MRI and correlates the imaging findings with the pathologic findings. CONCLUSION: On imaging studies, the appearance of fat necrosis ranges from typically benign to worrisome for malignancy. Mammography is more specific than sonography, and emphasis should be placed on mammography in making the diagnosis of fat necrosis. In selected cases, MRI may be helpful in showing findings consistent with fat necrosis.

Insuficiencia renal aguda por picadura de hormigas del palo santo (pseudomyrmex spp) / Acute renal failure due to the bite of ants of palo santo (pseudomyrmex spp)

La Insuficiencia Renal Aguda (IRA) es una complicación inusual de la picadura por hormigas secundaria a un daño tóxico isquémico con Necrosis Tubular Aguda. Los 2 pacientes presentados fueron transferidos del trópico Cochabambino al Hospital Clínico Viedma, experimentaron una reacción alérgica, hemólisis intravascular, rabdomiólisis e IRA posterior a un masivo ataque de las hormigas que habitan en el tronco del "Palo Santo" al que fueron atados por varias horas. Se dispone de pocos reportes acerca del compromiso renal en estos casos, así como de los mecanismos patogénicos implicados. Gracias al apoyo dialítico oportuno los pacientes mejoraron su condición clínica recuperando ambos la función renal en el lapso de 2 semanas de iniciada la hemodiálisis.

Acute Renal Insufficiency (ARI) is an unusual complication caused by the puncture of ants that falls into a secondary ischemic toxic damage with an Acute Tubular Necrosis. The two patients presented were transferred from the tropic of Cochabamba state to the Viedma Hospital, they have experienced an allergic reaction, intravascular hemolysis, rhabdomyolysis and ARI caused by the massive attack of ants that inhabit the trunk of the "Palo Santo" where they were tied by several hours. Few reports are known about the affectedness of the kidney organ, as well as the implied pathogenic mechanisms. The patients improved their clinical condition thanks to the opportune support; both recovered the kidney function after 2 weeks from the initiated hemodialysis.

Analysis of latent tuberculosis and mycobacterium avium infection data using mixture models.

BACKGROUND: Estimation of the frequency of latent tuberculosis infection (LTBI) is difficult in areas with low tuberculosis infection rates and high exposure to non-tuberculous mycobacteria (NTM), including BCG vaccination. The objective was to assess LTBI and M avium infection and to estimate their probability based on skin tests responses in an infant population from a region with the aforementioned characteristics. METHODS: A population-based tuberculin skin test (TST) and sensitin (M avium) survey was conducted on seven years old infants in Biscay, a province from The Basque Country (Spain). 2268 schoolchildren received sensitin and 5277 TST. Participation rate was 89%. Commonly used estimation methods were compared with a method based on the fit of mixture models using the Expectation Maximization algorithm. Functions estimating the probabilities of LTBI and M avium infection given the observed skin tests responses were developed for vaccinated and unvaccinated children. RESULTS: LTBI prevalences varied widely according to the estimation method. The mixture model provided prevalences higher than expected although intermediates between those obtained by currently recommended approaches. Exposure to previous BCG vaccine produces an upward shift of an average of about 3 mm on the induration size to attain the same probability of infection. CONCLUSION: Our results confirm the commonplace exposure to NTM which effect should be taken into account when performing and assessing tuberculin surveys. The use of mixture analysis under the empirical Bayes framework allows to better estimate the probability of LTBI in settings with presence of other NTM and high BCG-vaccination coverage. An estimation of the average effect of BCG vaccination on TST induration is also provided. These models maximise information coming from classical tuberculin surveys and could be used together with the newly developed blood tests to improve survey's specificity and cost-effectiveness.