Strong-LAMP Assay Based on a Strongyloides spp.-Derived Partial Sequence in the 18S rRNA as Potential Biomarker for Strongyloidiasis Diagnosis in Human Urine Samples

Human strongyloidiasis a soil-transmitted infection caused by Strongyloides stercoralis is one of the most neglected amongst the so-called Neglected Tropical Diseases (NTDs). S. stercoralis is a nematode, which is distributed worldwide; it has been estimated that it could affect millions of people, mainly in tropical and subtropical endemic regions. The difficulties of diagnosis lead to infection rates being underreported. Asymptomatic patients have chronic infections that can lead to severe hyperinfection syndrome or disseminated strongyloidiasis in immunocompromised patients. Strongyloidiasis can easily be misdiagnosed because conventional faecal-based techniques lack of sensitivity for the morphological identification of infective larvae in faeces. None of the currently used molecular methods have used urine samples as an alternative to faecal samples for diagnosing strongyloidiasis. This study was thus aimed at comparing, for the first time, the use of a new loop-mediated isothermal amplification (LAMP) molecular assay (Strong-LAMP) to traditional methods on patients' urine samples. Twenty-four urine samples were taken from patients included in a study involving two Spanish hospitals for strongyloidiasis screening using parasitological and serological tests. Strongyloides larvae were found in 11 patients' faecal samples, thereby ascertaining that they had the disease. Other patients had high antibody titres but no larvae were found in their faeces. All urine samples were analysed by PCR and Strong-LAMP assay. No amplification occurred when using PCR. Strong-LAMP led to detecting S. stercoralis DNA in urine samples from patients having previously confirmed strongyloidiasis by parasitological tests and/or a suspicion of being infected by serological ones. The Strong-LAMP assay is a useful molecular tool for research regarding strongyloidiasis in human urine samples. After further validation, the Strong-LAMP assay could also be used for complementary and effective diagnosis of strongyloidiasis in a clinical setting

Triploidía (69 xxx): reporte de caso

Las aberraciones cromosómicas son anormalidades que se presentan en el número o estructura de los cromosomas. En la literatura se han descrito alrededor de 4.000 enfermedades o síndromes relacionados con las anomalías congénitas que afectan alrededor del 3% de recién nacidos y son la causa más frecuente de mortalidad neonatal; adicionalmente generan discapacidad infantil y problemas psicosociales principalmente en países en vía de desarrollo. La triploidía 69 es una alteración numérica, se estima que entre el 1 al 3% de todas las concepciones humanas tendrían embriones triploides y que de éstas el 99,9% se pierden entre el primer y segundo trimestre, 15% de los fetos terminan en abortos espontáneos antes de las 20 semanas de gestación. El propósito del trabajo es describir un caso de triploidía 69 en líquido amniótico que terminó en muerte fetal. Abstract Chromosomal aberrations are abnormalities that are present in the number or structure of chromosomes; in the literature about 4,000 diseases or syndromes related to congenital anomalies have been described, affecting about 3% of newborns and are the most frequent cause of neonatal mortality; additionally generate children’s disability and psychosocial problems mainly in developing countries. The triploidy 69 is a numerical alteration, it is estimated that between 1 and 3% of all human conceptions would have triploid embryos and of these 99.9% are lost between the first and second trimester, 15% of the fetuses end in spontaneous abortions before 20 weeks of gestation. The purpose of our work is to describe a case of triploidy 69 in amniotic fluid that ended in fetal death. &nbsp

Triploidía (69 xxx): reporte de caso

Las aberraciones cromosómicas son anormalidades que se presentan en el número o estructura de los cromosomas. En la literatura se han descrito alrededor de 4.000 enfermedades o síndromes relacionados con las anomalías congénitas que afectan alrededor del 3% de recién nacidos y son la causa más frecuente de mortalidad neonatal; adicionalmente generan discapacidad infantil y problemas psicosociales principalmente en países en vía de desarrollo. La triploidía 69 es una alteración numérica, se estima que entre el 1 al 3% de todas las concepciones humanas tendrían embriones triploides y que de éstas el 99,9% se pierden entre el primer y segundo trimestre, 15% de los fetos terminan en abortos espontáneos antes de las 20 semanas de gestación. El propósito del trabajo es describir un caso de triploidía 69 en líquido amniótico que terminó en muerte fetal. Abstract Chromosomal aberrations are abnormalities that are present in the number or structure of chromosomes; in the literature about 4,000 diseases or syndromes related to congenital anomalies have been described, affecting about 3% of newborns and are the most frequent cause of neonatal mortality; additionally generate children’s disability and psychosocial problems mainly in developing countries. The triploidy 69 is a numerical alteration, it is estimated that between 1 and 3% of all human conceptions would have triploid embryos and of these 99.9% are lost between the first and second trimester, 15% of the fetuses end in spontaneous abortions before 20 weeks of gestation. The purpose of our work is to describe a case of triploidy 69 in amniotic fluid that ended in fetal death. &nbsp

The supporting role of the teres major muscle, an additional component in glenohumeral stability? An anatomical and radiological study

Muscle coordination plays an important role in glenohumeral stability. The rotator cuff and the long head of the biceps are considered the primary dynamic stabilizers muscles. However, the fact that a subgroup of patients with a massive tear in the rotator cuff were able to keep a normal function, should make us question this traditional view. We hypothesize that the teres major which is also a monoarticular scapulohumeral muscle, although it is not part of the conjoined tendon of the rotator cuff, can play a role in glenohumeral stability by a direct support of the humeral head generated by the particular posteroanterior location of this muscle under the humeral head and which, as far as we know, has not been written up previously. This particular effect could appear while the arm is being lifted and the humeral head could be leaning on against the teres major muscle belly underneath it. An anatomical a radiological study was carried out to substantiate our hypothesis. Two cadaver specimens were used for the anatomical study. Frist body was studied through conventional dissection. The second body was analysed through sectional anatomy. Then a radiological study was carried out using magnetic resonance imaging in a healthy male volunteer. Both anatomically and radiologically, the anteroinferior surface of the humeral head was showed firmly resting against the muscle belly of the teres major, to the point of misshaping it from 110 degrees of arm elevation with external rotation. The specific contribution of this effect to the glenohumeral stability needs to be confirmed by further studies and can help us to prevent the high incidence of glenohumeral dislocations

Is dry needling of the supinator a safe procedure? A potential treatment for lateral epicondylalgia or radial tunnel syndrome. A cadaveric study

The supinator muscle is involved in two pain conditions of the forearm and wrist: lateral epicondylalgia and radial tunnel syndrome. Its close anatomical relationship with the radial nerve at the arcade of Frohse encourages research on dry needling approaches. Our aim was to determine if a solid filiform needle safely penetrates the supinator muscle during the clinical application of dry needling. Needle insertion of the supinator muscle was conducted in ten cryopreserved forearm specimens with a 30 × 0.32 mm filiform needle. With the forearm pronated, the needle was inserted perpendicular into the skin at the dorsal aspect of the forearm at a point located 4cm distal to the lateral epicondyle. The needle was advanced to a depth judged to be in the supinator muscle. Safety was assessed by measuring the distance from the needle to the surrounding neurovascular bundles of the radial nerve. Accurate needle penetration of the supinator muscle was observed in 100% of the forearms (needle penetration:16.4 ± 2.7 mm 95% CI 14.5 mm to 18.3 mm). No neurovascular bundle of the radial nerve was pierced in any of the specimen’s forearms. The distances from the tip of the needle were 7.8 ± 2.9 mm (95% CI 5.7 mm to 9.8 mm) to the deep branch of the radial nerve and 8.6 ± 4.3 mm (95% CI 5.5 mm to 11.7 mm) to the superficial branch of the radial nerve. The results from this cadaveric study support the assumption that needling of the supinator muscle can be accurately and safely conducted by an experienced clinician

Safety of Dry Needling of the Pronator Teres Muscle in Cadavers: A Potential Treatment for Pronator Syndrome

Background: Entrapment of the median nerve at the pronator teres muscle can contribute to symptoms in the forearm and wrist. The pronator teres is also involved in patterns of spasticity observed in people who had suffered a stroke. Research on treatment efficacy with dry needling is scarce. Objective: To determine if a solid filiform needle safely penetrates the pronator teres muscle during the clinical application of dry needling. Design: A cadaveric descriptive study. Methods: Needle insertion of the pronator teres was conducted in ten cryopreserved forearms with a 30*0.32 mm filiform needle. With the forearm supinated, the needle was inserted 3 cm distal to the mid-point between the biceps tendon insertion and the medial epicondyle. The needle was advanced in a cranial and medial direction to a depth clinically judged to be in the pronator teres muscle. Safety was assessed by measuring the distance from the needle to the surrounding neurovascular bundles. Results: Accurate needle penetration of the pronator teres was observed in 100% of the specimens (mean needle penetration: 16.7 ± 4.3 mm, 95%CI 13.6 to 19.7 mm). No neurovascular bundles were pierced in any of the specimen's forearms. The distances from the tip of the needle to the surrounding neurovascular bundles were 16.4 ± 3.9 mm (95%CI 13.6 to 19.2 mm) to the ulnar nerve (A), 9.0 ± 2.2 mm (95%CI 7.3 to 19.5 mm) to the median nerve (B), and 12.8 ± 4.0 mm (95%CI 10.0 to 15.7 mm) to brachial artery (C). Conclusion: The results from this cadaveric study support the assumption that needling of the pronator teres using described anatomical landmarks can be accurately and safely conducted by an experienced clinician

Congenital leptin deficiency and leptin gene missense mutation found in two colombian sisters with severe obesity

Background: Congenital leptin deficiency is a recessive genetic disorder associated with severe early-onset obesity. It is caused by mutations in the leptin (LEP) gene, which encodes the protein product leptin. These mutations may cause nonsense-mediated mRNA decay, defective secretion or the phenomenon of biologically inactive leptin, but typically lead to an absence of circulating leptin, resulting in a rare type of monogenic extreme obesity with intense hyperphagia, and serious metabolic abnormalities. Methods: We present two severely obese sisters from Colombia, members of the same lineal consanguinity. Their serum leptin was measured by MicroELISA. DNA sequencing was performed on MiSeq equipment (Illumina) of a next-generation sequencing (NGS) panel involving genes related to severe obesity, including LEP. Results: Direct sequencing of the coding region of LEP gene in the sisters revealed a novel homozygous missense mutation in exon 3 [NM_002303.3], C350G>T [p.C117F]. Detailed information and clinical measurements of these sisters were also collected. Their serum leptin levels were undetectable despite their markedly elevated fat mass. Conclusions: The mutation of LEP, absence of detectable leptin, and the severe obesity found in these sisters provide the first evidence of monogenic leptin deficiency reported in the continents of North and South America. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Role of LCN2 in a murine model of hindlimb ischemia and in peripheral artery disease patients, and its potential regulation by miR-138-5P

Background and aims: Peripheral arterial disease (PAD) is a leading cause of morbimortality worldwide. Lipocalin-2 (LCN2) has been associated with higher risk of amputation or mortality in PAD and might be involved in muscle regeneration. Our aim is to unravel the role of LCN2 in skeletal muscle repair and PAD.Methods and results: WT and Lcn2(-/-) mice underwent hindlimb ischemia. Blood and crural muscles were analyzed at the inflammatory and regenerative phases. At day 2, Lcn2(-/-) male mice, but not females, showed increased blood and soleus muscle neutrophils, and elevated circulating pro-inflammatory monocytes (p < 0.05), while locally, total infiltrating macrophages were reduced (p < 0.05). Moreover, Lcn2(-/-) soleus displayed an elevation of Cxcl1 (p < 0.001), and Cxcr2 (p < 0.01 in males), and a decrease in Ccl5 (p < 0.05). At day 15, Lcn2 deficiency delayed muscle recovery, with higher density of regenerating myocytes (p < 0.04) and arterioles (alpha SMA(+), p < 0.025). Reverse target prediction analysis identified miR-138-5p as a potential regulator of LCN2, showing an inverse correlation with Lcn2 mRNA in skeletal muscles (rho = -0.58, p < 0.01). In vitro, miR-138-5p mimic reduced Lcn2 expression and luciferase activity in murine macrophages (p < 0.05). Finally, in human serum miR-138-5p was inversely correlated with LCN2 (p <= 0.001 adjusted, n = 318), and associated with PAD (Odds ratio 0.634, p = 0.02, adjusted, PAD n = 264, control n = 54).Conclusions: This study suggests a possible dual role of LCN2 in acute and chronic conditions, with a probable role in restraining inflammation early after skeletal muscle ischemia, while being associated with vascular damage in PAD, and identifies miR-138-5p as one potential post-transcriptional regulator of LCN2.Vascular Surger

Continuous cultivation of photosynthetic microorganisms: approaches, applications and future trends

The possibility of using photosynthetic microorganisms, such as cyanobacteria and microalgae, for converting light and carbon dioxide into valuable biochemical products has raised the need for new cost-efficient processes ensuring a constant product quality. Food, feed, biofuels, cosmetics and pharmaceutics are among the sectors that can profit from the application of photosynthetic microorganisms. Biomass growth in a photobioreactor is a complex process influenced by multiple parameters, such as photosynthetic light capture and attenuation, nutrient uptake, photobioreactor hydrodynamics and gas-liquid mass transfer. In order to optimize productivity while keeping a standard product quality, a permanent control of the main cultivation parameters is necessary, where the continuous cultivation has shown to be the best option. However it is of utmost importance to recognize the singularity of continuous cultivation of cyanobacteria and microalgae due to their dependence on light availability and intensity. In this sense, this review provides comprehensive information on recent breakthroughs and possible future trends regarding technological and process improvements in continuous cultivation systems of microalgae and cyanobacteria, that will directly affect cost-effectiveness and product quality standardization. An overview of the various applications, techniques and equipment (with special emphasis on photobioreactors) in continuous cultivation of microalgae and cyanobacteria are presented. Additionally, mathematical modelling, feasibility, economics as well as the applicability of continuous cultivation into large-scale operation, are discussed.This research work was supported by the grant SFRH/BPD/98694/2013 (Bruno Fernandes) from Fundacao para a Ciencia e a Tecnologia (Portugal). The authors thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013. The authors also thank the Project "BioInd Biotechnology and Bioengineering for improved Industrial and Agro-Food processes, REF. NORTE-07-0124-FEDER-000028" Co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDE