Influence of state-of-the-art laboratory techniques on the phenotyping of women with polycystic ovary syndrome in the clinical setting.

PURPOSE: Evidence-based guidelines for the management of polycystic ovary syndrome (PCOS) recommend clinical laboratories use liquid chromatography-tandem mass spectrometry (LC-MS/MS) for diagnosing biochemical hyperandrogenism. However, automated immunoassays are still mostly used in routine laboratories worldwide. Another hurdle for PCOS phenotyping in the clinical setting is ultrasound assessment of polycystic ovarian morphology. We address the impact of using state-of-the-art (LC-MS/MS) and of an anti-müllerian hormone (AMH) assay on the diagnosis of PCOS in routine practice. METHODS: In a cross-sectional study, we included 359 premenopausal women consecutively evaluated because of symptoms of functional androgen excess or hyperandrogenemia, and finally diagnosed with PCOS. Patients were submitted to routine phenotyping based on serum androgen measurements by immunoassays and an ovarian ultrasound when necessary. Samples of all patients were also assayed by LC-MS/MS for hyperandrogenemia and for circulating AMH. RESULTS: The observed agreement between immunoassays and LC-MS/MS in identifying hyperandrogenemia was poor [78.0%; k(95%CI): 0.366 (0.283;0.449)]. The observed agreement between ultrasound and increased AMH was 27.3% [(95%CI): 0.060 (0.005; 0.115)]. Using LC-MS/MS changed PCOS phenotypes in 60(15.8%) patients. Fifty-two (18.3%) individuals with hyperandrogenemia by routine immunoassays no longer presented with androgen excess by LC-MS/MS. Overall diagnostic agreement between routine assessment using immunoassays and ultrasound and that derived from LC-MS/MS and the addition of AMH to US was moderate [weighted κ (linear weights): 0.512 (0.416;0.608)]. CONCLUSIONS: Immunoassays used in routine practice are unacceptably inaccurate for phenotyping women with PCOS. Our data cast some doubts upon the interchangeability of serum AMH and ultrasound examination for the diagnosis of PCOS.

Postmenopausal onset of androgen excess: a diagnostic and therapeutic algorithm based on extensive clinical experience.

PURPOSE: Postmenopausal hyperandrogenism is a rare condition that requires identifying those women bearing a life-threatening tumor. We aimed to study diagnostic work-up and management of postmenopausal androgen excess, proposing an algorithm for clinical decision supporting. METHODS: We conducted an observational cross-sectional study and longitudinal follow-up including 51 consecutive menopausal patients reported for hyperandrogenism between 2003 and 2023 to our clinics. We assessed diagnostic testing accuracy and performance by receiver operating characteristic curves, their respective areas under the curve (AUCROC), and 95% confidence intervals (95%CI), for distinguishing between benign and malignant conditions, and androgen excess source. RESULTS: Most commonly, postmenopausal hyperandrogenism derived from benign conditions such as ovarian hyperthecosis (n = 9). However, four (8%) patients had borderline/malignant tumors arising at the ovaries (n = 3) or adrenals (n = 1). These latter were more likely to develop virilization than those with benign disorders [specificity(95%CI)]: 0.87 (0.69; 0.92)]. Circulating total testosterone [AUCROC(95%CI): 0.899 (0.795; 1.000)] and estradiol [AUCROC(95%CI): 0.912 (0.812; 1.000)] concentrations showed good performances for discriminating between both conditions. Transvaginal-ultrasonography found two out of three potentially malignant ovarian neoplasms, and another was apparent on a pelvic computed tomography scan. An adrenal computed tomography scan also located an androgen-secreting carcinoma. CONCLUSIONS: Clinical or biochemical features of an aggressive androgen-secreting tumor should lead to urgently obtaining a targeted imaging. At first, an abdominal-pelvic CT scan represents the best choice to perceive adrenal malignancy, and may identify aggressive ovarian tumors. When warning signs are lacking, a calm and orderly work-up allows properly addressing the diagnostic challenge of postmenopausal hyperandrogenism.

[Influence of constraint-induced movement therapy on childhood hemiparesis with moderate manual dexterity at 16 years of age: a systematic review]. / Influencia de la terapia de movimiento inducido por restricción en hemiparesia infantil con habilidad manual moderada de 2 a 16 años: revisión sistemática.

INTRODUCTION: The deficit of use on the upper limb affects, as well as the quality of movement, influences the execution of a functional grasp (action of grasping-releasing an object) and therefore, reduces the possibilities of execution of the child's daily activities diagnosed with infantile hemiplegia. Manual ability (finger movements, global and distal grasp, handling, precise movement, fluid...) could be a factor that determines whether the execution of constraint induced movement therapy favors the increase in functionality of the upper limb affected. AIM: To study the constraint induced movement therapy influence on the functionality of the affected upper limb in infantile hemiplegia with moderate manual ability from 2 to 16 years of age. PATIENTS AND METHODS: A bibliographic search of the works published between 2014 and 2019 was performed in the PubMed, PEDro and The Cochrane Library databases. RESULTS: After applying the inclusion/exclusion criteria, seven studies were obtained from a total of 203 to be analyzed, comparing constraint induced movement therapy with other interventions. CONCLUSIONS: The results comparison is complex due to the lack of unanimity in the doses application and evaluation tools of the affected segment. The exclusive application of constraint induced movement therapy allows greater benefits in functionality, at the level of manual ability on affected upper limb in infantile hemiplegia with moderate manual ability than conventional therapy or other therapies.

TITLE: Influencia de la terapia de movimiento inducido por restricción en hemiparesia infantil con habilidad manual moderada de 2 a 16 años: revisión sistemática.Introducción. El déficit de uso de la extremidad superior afectada, así como la calidad del movimiento, influyen en la ejecución de un agarre funcional (acción de agarrar-liberar un objeto) y, por tanto, reducen las posibilidades de ejecución de las actividades cotidianas del niño diagnosticado de hemiplejía infantil. La habilidad manual (movimientos de los dedos, agarre global y distal, manipulación, movimiento preciso, fluido…) podría ser un factor que determine si la ejecución de terapia de movimiento inducido por restricción favorece el incremento de la funcionalidad de la extremidad superior afectada. Objetivo. Estudiar la influencia de la terapia de movimiento inducido por restricción en la funcionalidad de la extremidad superior afectada en hemiplejía infantil con una habilidad manual moderada de 2 a 16 años. Pacientes y métodos. Se realizó una búsqueda bibliográfica de los trabajos publicados entre 2014 y 2019 en las bases de datos PubMed, PEDro y The Cochrane Library. Resultados. Tras aplicar los criterios de inclusión y exclusión, se obtuvieron siete estudios, de un total de 203, para ser analizados, en los que se compara la terapia de movimiento inducido por restricción con otras intervenciones. Conclusiones. La comparativa de resultados resulta compleja debido a la falta de unanimidad en la aplicación de dosis y herramientas de evaluación del segmento afectado. La terapia de movimiento inducido por restricción aplicada de manera exclusiva permite mayores beneficios en la funcionalidad en cuanto a destreza manipulativa de la extremidad superior afectada en hemiplejía infantil con habilidad manual moderada que la terapia convencional u otras terapias.

Nox1-derived oxidative stress as a common pathogenic link between obesity and hyperoxaluria-related kidney injury.

Specific relationships among reactive oxygen species, activation pathways, and inflammatory mechanisms involved in kidney injury were assessed in a combined model of obesity and hyperoxaluria. Male Wistar rats were divided into four groups: Control, HFD (high fat diet), OX (0.75% ethylene glycol), and HFD + OX (combined model) Changes in basal O2- levels were evaluated by chemiluminescence in renal interlobar arteries and renal cortex. Furthermore, the effect of different inhibitors on NADPH-stimulated O2- generation was assessed in renal cortex. Oxidative stress sources, and local inflammatory mediators, were also determined, in parallel, by RT-PCR, and correlated with measures of renal function, urinary biochemistry, and renal structure. Rats from the HFD group developed overweight without lipid profile alteration. Tubular deposits of crystals were seen in OX and severely enhanced in HFD + OX groups along with a significantly higher impairment of renal function. Basal oxidative stress was increased in renal cortex of OX rats and in renal arteries of HFD rats, while animals from the combined HFD + OX group exhibited the highest levels of oxidative stress in renal cortex, derived from xanthine oxidase and COX-2. NADPH oxidase-dependent O2- generation was elevated in renal cortex of the OX group and markedly enhanced in the HFD + OX rats, and associated to an up-regulation of Nox1 and a down-regulation of Nox4 expression. High levels of oxidative stress in the kidney, of OX and HFD + OX groups were also associated to an inflammatory response mediated by an elevation of TNFα, COX-2, NFκB1 MCP-1, and OPN. Oxidative stress is a key pathogenic factor in renal disease associated to hyperoxaluria and a common link underlying the exacerbated inflammatory response and kidney injury found under conditions of both obesity and hyperoxaluria. Nox1 pathway must be considered as a potential therapeutic target.

Anti-inflammatory effects of aloe vera on soy meal-induced intestinal inflammation in zebrafish.

Soybean meal is one of the most promising alternatives to replace fishmeal in the aquaculture industry. However, its ingestion triggers an intestinal inflammatory process that compromises fish health and nutrition. Therefore, finding strategies that reduce the deleterious effects of a soy protein-based diet are relevant. In this work we analyzed the effects of an aloe vera (Aloe barbadensis miller, AV) extract on intestinal inflammation and innate immunity of zebrafish by adding it to the water and by supplementing it in a soybean meal-based diet. To search for potential immunomodulatory effects of AV, we tested its effectiveness in two inflammation assays and compared fish fed with either fishmeal or soybean meal-based feed supplemented with AV. Our results show a strong anti-inflammatory effect of AV. Furthermore, while soy-based meal strongly induces the expression of inflammation markers, supplementation with AV reverted this effect. Finally, we show that fish fed with a soy meal diet are highly susceptible to bacterial infection, but that this condition is significantly reduced when the soy meal is supplemented with AV. Our results suggest that AV is a good candidate to be incorporated as an additive in farmed fish diets to facilitate the replacement of fishmeal by soybean meal, maintaining intestinal health.

Junctional scotoma. A case report. / Escotoma juncional. A propósito de un caso.

The case concerns a 55 year-old female patient with progressive visual decrease in her right eye that showed a junctional scotoma in the visual field study. A magnetic resonance scan was performed, which showed a right middle cerebral artery aneurysm with ipsolateral chiasmatic involvement.

NETs detection and quantification in paraffin embedded samples using confocal microscopy.

Detection and quantification of Neutrophil Extracellular Traps (NETs) in tissue samples has become a topic of great interest to understand their pathological role in various diseases. We describe a semi-automatic method of visualization and quantification of NETs in paraffin-embedded intracoronary thrombus aspirate samples. This study is based on colocalization of myeloperoxidase (MPO) and citrullinated histone 3 (H3Cit) as hallmark of the presence of NETs. For the analysis we used the confocal immunofluorescence microscopy technology to quantify the number of fields and the total area (in µm2) containing NETs in each thrombus sample. This observer-independent quantification method could be a useful tool to standardize the study of NETs in paraffin-embedded tissues, enabling comparison of results among different laboratories.

Autoimmune neuroretinopathy secondary to Zika virus infection. / Neurorretinopatía autoinmune secundaria a infección por el virus del Zika.

CASE REPORT: A 40-year-old woman diagnosed with Zika virus infection 6 months before she arrived at this hospital. She referred to a progressive and painless vision loss, of 2 weeks onset after the infection diagnosis. She was treated with topical steroids. Previous visual acuity was recovered, but she still refers to reduced visual field and nyctalopia. Ophthalmologic examination revealed severe retinal sequels, compatible with autoimmune retinopathy. Based on the clinical features and the temporal relationship with Zika virus infection, non-para-neoplastic autoimmune retinopathy was diagnosed and managed with steroids and infliximab. DISCUSSION: Zika virus can trigger a non-para-neoplastic autoimmune retinopathy. The diagnosis is based on clinical features, and requires early immunosuppressive therapy.

Metabolic syndrome contributes to renal injury mediated by hyperoxaluria in a murine model of nephrolithiasis.

Metabolic syndrome (MS) individuals have a higher risk of developing chronic kidney disease through unclear pathogenic mechanisms. MS has been also related with higher nephrolithiasis prevalence. To establish the influence of MS on renal function, we designed a murine model of combined metabolic syndrome and hyperoxaluria. Four groups of male Sprague-Dawley rats were established: (1) control group (n = 10) fed with standard chow; (2) stone former group (SF) (n = 10) fed with standard chow plus 0.75% ethylene glycol administered in the drinking water; (3) metabolic syndrome group (MS) (n = 10), fed with 60% fructose diet; (4) metabolic syndrome + stone former group (MS + SF) (n = 10), 60% fructose diet and 0.75% EG in the drinking water. MS group showed a significant injury to renal function when hyperoxaluria was induced. It was demonstrated by a significant decrease of creatinine clearance (p < 0.001), with higher tubular damage (34.3%, CI 95% 23.9-44.7, p < 0.001), produced by deposition of crystals, and increased tubular synthesis of osteopontin as a response to tubular damage. Induction of hyperoxaluria in rats with MS causes severe morphological alterations with a significant impairment of renal function. This impairment is not produced in rats without MS. Therefore, this model can be useful for the study of the influence of MS in stone formation.

Topical imiquimod 5% as an alternative therapy in periocular basal cell carcinoma in two patients with surgical contraindication. / Imiquimod 5% tópico como terapia alternativa en carcinoma basocelular periocular en 2 pacientes con contraindicación quirúrgica.

CLINICAL CASES: The cases are presented of two patients with periocular basal cell carcinoma of the eyelid who received topical imiquimod 5%, with a good response. Both had a functional state that contraindicated surgical treatment. CONCLUSION: Imiquimod cream 5% was shown to be an effective alternative to surgical treatment of periocular basal cell carcinoma, especially in those cases where surgery is not possible.

Parinaud's oculoglandular syndrome: A case report. / Síndrome oculoglandular de Parinaud: a propósito de un caso.

CLINICAL CASE: A 33-year old woman presents with unilateral granulomatous conjunctivitis, ipsilateral regional lymphadenopathy and fever. A Bartonella henselae infection is demonstrated by indirect immunofluorescence, and a diagnosis of a Parinaud's oculoglandular syndrome is established. Outcome after treatment with oral doxycycline is satisfactory. DISCUSSION: Parinaud's oculoglandular syndrome is the most frequent ocular manifestation of a Bartonella henselae infection.

Active site mapping of Loxosceles phospholipases D: Biochemical and biological features.

Brown spider phospholipases D from Loxosceles venoms are among the most widely studied toxins since they induce dermonecrosis, triggering inflammatory responses, increase vascular permeability, cause hemolysis, and renal failure. The catalytic (H12 and H47) and metal-ion binding (E32 and D34) residues in Loxosceles intermedia phospholipase D (LiRecDT1) were mutated to understand their roles in the observed activities. All mutants were identified using whole venom serum antibodies and a specific antibody to wild-type LiRecDT1, they were also analyzed by circular dichroism (CD) and differential scanning calorimetry (DSC). The phospholipase D activities of H12A, H47A, H12A-H47A, E32, D34 and E32A-D34A, such as vascular permeability, dermonecrosis, and hemolytic effects were inhibited. The mutant Y228A was equally detrimental to biochemical and biological effects of phospholipase D, suggesting an essential role of this residue in substrate recognition and binding. On the other hand, the mutant C53A-C201A reduced the enzyme's ability to hydrolyze phospholipids and promote dermonecrosis, hemolytic, and vascular effects. These results provide the basis understanding the importance of specific residues in the observed activities and contribute to the design of synthetic and specific inhibitors for Brown spider venom phospholipases D.

[External cephalic version]. / Versión cefálica externa.

Objetives: To analyze the rate of successful external cephalic versions in our center and caesarean sections that would be avoided with the use of external cephalic versions. Material and methods: From January 2012 to March 2016 external cephalic versions carried out at our center, which were a total of 52. We collected data about female age, gestational age at the time of the external cephalic version, maternal body mass index (BMI), fetal variety and situation, fetal weight, parity, location of the placenta, amniotic fluid index (ILA), tocolysis, analgesia, and newborn weight at birth, minor adverse effects (dizziness, hypotension and maternal pain) and major adverse effects (tachycardia, bradycardia, decelerations and emergency cesarean section). Results: 45% of the versions were unsuccessful and 55% were successful. The percentage of successful vaginal delivery in versions was 84% (4% were instrumental) and 15% of caesarean sections. With respect to the variables studied, only significant differences in birth weight were found; suggesting that birth weight it is related to the outcome of external cephalic version. Probably we did not find significant differences due to the number of patients studied. Conclusion: For women with breech presentation, we recommend external cephalic version before the expectant management or performing a cesarean section. The external cephalic version increases the proportion of fetuses in cephalic presentation and also decreases the rate of caesarean sections.

Exosomes enriched in stemness/metastatic-related mRNAS promote oncogenic potential in breast cancer.

Cancer cells efficiently transfer exosome contents (essentially mRNAs and microRNAs) to other cell types, modifying immune responses, cell growth, angiogenesis and metastasis. Here we analyzed the exosomes release by breast tumor cells with different capacities of stemness/metastasis based on CXCR4 expression, and evaluated their capacity to generate oncogenic features in recipient cells. Breast cancer cells overexpressing CXCR4 showed an increase in stemness-related markers, and in proliferation, migration and invasion capacities. Furthermore, recipient cells treated with exosomes from CXCR4-cells showed increased in the same abilities. Moreover, inoculation of CXCR4-cell-derived exosomes in immunocompromised mice stimulated primary tumor growth and metastatic potential. Comparison of nucleic acids contained into exosomes isolated from patients revealed a "stemness and metastatic" signature in exosomes of patients with worse prognosis. Finally, our data supported the view that cancer cells with stem-like properties show concomitant metastatic behavior, and their exosomes stimulate tumor progression and metastasis. Exosomes-derived nucleic acids from plasma of breast cancer patients are suitable markers in the prognosis of such patients.

Using mixture design of experiments to assess the environmental impact of clay-based structural ceramics containing foundry wastes.

This work describes the leaching behavior of potentially hazardous metals from three different clay-based industrial ceramic products (wall bricks, roof tiles, and face bricks) containing foundry sand dust and Waelz slag as alternative raw materials. For each product, ten mixtures were defined by mixture design of experiments and the leaching of As, Ba, Cd, Cr, Cu, Mo, Ni, Pb, and Zn was evaluated in pressed specimens fired simulating the three industrial ceramic processes. The results showed that, despite the chemical, mineralogical and processing differences, only chrome and molybdenum were not fully immobilized during ceramic processing. Their leaching was modeled as polynomial equations, functions of the raw materials contents, and plotted as response surfaces. This brought to evidence that Cr and Mo leaching from the fired products is not only dependent on the corresponding contents and the basicity of the initial mixtures, but is also clearly related with the mineralogical composition of the fired products, namely the amount of the glassy phase, which depends on both the major oxides contents and the firing temperature.

Structural Insights into Substrate Binding of Brown Spider Venom Class II Phospholipases D.

Phospholipases D (PLDs), the major dermonecrotic factors from brown spider venoms, trigger a range of biological reactions both in vitro and in vivo. Despite their clinical relevance in loxoscelism, structural data is restricted to the apo-form of these enzymes, which has been instrumental in understanding the functional differences between the class I and II spider PLDs. The crystal structures of the native class II PLD from Loxosceles intermedia complexed with myo-inositol 1-phosphate and the inactive mutant H12A complexed with fatty acids indicate the existence of a strong ligand-dependent conformation change of the highly conserved aromatic residues, Tyr 223 and Trp225 indicating their roles in substrate binding. These results provided insights into the structural determinants for substrate recognition and binding by class II PLDs.

Effect of intratympanic dimethyl sulphoxide (DMSO) in an in vivo model of cisplatin-related ototoxicity.

Although dimethyl sulfoxide (DMSO) is one of the most common solvents employed in otoprotection studies, its effect on the inner ear remains unknown. Only a few in vitro studies have addressed the effect of DMSO in cochlear cells. Up to the date, no in vivo functional studies have been reported. To determine the effect of intratympanic DMSO application in the inner ear, and to evaluate its effect in combination with cisplatin in Wistar rats, twelve Wistar rats were randomly assigned into two groups. Group A received intratympanic 1 % DMSO in both ears. Group B received intraperitoneal cisplatin (10 mg/kg) and intratympanic 0.5 % DMSO in the right ear and saline solution in the left ear. Functional changes were evaluated with Auditory Steady-State Responses before and 5 days after the procedure. Morphological changes were studied by means of confocal laser scanning microscopy following the removal of the temporal bones and cochlear dissection. Hearing threshold levels in group A did not show any statistically significant changes after the treatment. In group B, significant differences between pre- and post-treatment were found, with no statistically significant variations between right (DMSO) and left ear (saline solution). We suggest that DMSO could be safely used to dissolve hydrophobic compounds in otoprotection studies without interfering with the cochlear damage produced by cisplatin.

Effects of seed burial on germination, protein mobilisation and seedling survival in Dodonaea viscosa.

Ecological restoration of disturbed areas requires substantial knowledge of the germination of native plants and the creation of novel methods to increase seedling establishment in the field. We studied the effects of soil matrix priming on the germination of Dodonaea viscosa seeds, which exhibit physical dormancy. To this end, we buried both pre-scarified (in H2SO4, 3 min) and non-pre-scarified seeds in the Parque Ecológico de la Ciudad de México. After seeds were unearthed, they were post-scarified for 0, 2, 6 and 10 min and their germination percentages compared to the germination of a control batch of laboratory-stored seeds. For both control and unearthed seeds, the protein pattern was determined in the enriched storage protein fraction in SDS-PAGE gels stained with Coomassie blue. Percentage germination increased as the scarification time increased. Pre-scarification significantly increased percentage germination of post-scarified seeds in relation to the control and non-pre-scarified seeds. In seeds unearthed from the forest site, the buried pre-scarified seeds had relatively high percentage germination, even in the absence of post-scarification treatment. A 48-kDa protein was not found in unearthed, pre-scarified seeds nor in the control germinated seeds, indicating that mobilisation of this protein occurred during soil priming. Burying seeds for a short period, including the beginning of the rainy season, promoted natural priming, which increased protein mobilisation. Functionally, priming effects were reflected in high percentage seedling survival in both the shade house and the field. Seed burial also reduced the requirement for acidic post-scarification.

Fractal continuum model for tracer transport in a porous medium.

A model based on the fractal continuum approach is proposed to describe tracer transport in fractal porous media. The original approach has been extended to treat tracer transport and to include systems with radial and uniform flow, which are cases of interest in geoscience. The models involve advection due to the fluid motion in the fractal continuum and dispersion whose mathematical expression is taken from percolation theory. The resulting advective-dispersive equations are numerically solved for continuous and for pulse tracer injection. The tracer profile and the tracer breakthrough curve are evaluated and analyzed in terms of the fractal parameters. It has been found in this work that anomalous transport frequently appears, and a condition on the fractal parameter values to predict when sub- or superdiffusion might be expected has been obtained. The fingerprints of fractality on the tracer breakthrough curve in the explored parameter window consist of an early tracer breakthrough and long tail curves for the spherical and uniform flow cases, and symmetric short tailed curves for the radial flow case.