Impact of heat waves on semen quality: A retrospective study in Argentina between 2005 and 2023.

Heat waves, defined as periods with daily temperatures surpassing the historical average for a specific region, have become more frequent worldwide in recent years. Previous studies have reported a negative association between temperature and semen quality, but the focus has mainly been on Asian and European populations. The study included 54,926 men (18-60 years) undergoing routine semen analysis between 2005 and 2023 at CEUSA-LAEH andrology unit, in Buenos Aires, Argentina. Hourly temperature readings were provided by the Servicio Meteorológico Nacional. R programming (R Studio v2022.07.2) was used to define heat waves, calculate key characteristics, visualize results, and perform statistical tests at the IBYME laboratory. During the period studied, a total of 124 days had heat waves (defined after at least 3 consecutive days with 32.3 °C and 22 °C). Men exposed to heat waves during spermatogenesis exhibited lower sperm number (concentration and count; P < 0.0001) and decreased normal morphology (percentage of normal sperm and normal motile count; P < 0.05) compared to those not exposed. These differences were most pronounced between semen samples from years with several heat waves (2013, 2023) and none (2005, 2007, 2016), displaying 4-5 times higher fold changes (P < 0.05). Further analysis employing multiple regression revealed a significantly negative association between semen quality and heat wave length, suggesting that a prolonged exposure may be more detrimental than an acute exposure. Subsequent analysis focusing on prolonged exposure (≥6-days heat wave) during spermatogenesis revealed a negative (P < 0.05) association between early exposure (spermatocytogenesis: 64-90 days prior semen collection) and semen quality. This study underscores the negative association between early exposure to heat waves during sperm development and semen quality, raising concerns about its possible association with the worldwide declining male fertility. A comprehensive collaborative approach is crucial, involving global governmental policies, sustainable practices, and coordinated efforts across scientific, healthcare, and policy domains.

Decellularized porcine peripheral nerve based injectable hydrogels as a Schwann cell carrier for injured spinal cord regeneration.

Objective.To develop a clinically relevant injectable hydrogel derived from decellularized porcine peripheral nerves and with mechanical properties comparable to native central nervous system (CNS) tissue to be used as a delivery vehicle for Schwann cell transplantation to treat spinal cord injury (SCI).Approach.Porcine peripheral nerves (sciatic and peroneal) were decellularized by chemical decellularization using a sodium deoxycholate and DNase (SDD) method previously developed by our group. The decellularized nerves were delipidated using dichloromethane and ethanol solvent and then digested using pepsin enzyme to form injectable hydrogel formulations. Genipin was used as a crosslinker to enhance mechanical properties. The injectability, mechanical properties, and gelation kinetics of the hydrogels were further analyzed using rheology. Schwann cells encapsulated within the injectable hydrogel formulations were passed through a 25-gauge needle and cell viability was assessed using live/dead staining. The ability of the hydrogel to maintain Schwann cell viability against an inflammatory milieu was assessedin vitrousing inflamed astrocytes co-cultured with Schwann cells.Mainresults. The SDD method effectively removes cells and retains extracellular matrix in decellularized tissues. Using rheological studies, we found that delipidation of decellularized porcine peripheral nerves using dichloromethane and ethanol solvent improves gelation kinetics and mechanical strength of hydrogels. The delipidated and decellularized hydrogels crosslinked using genipin mimicked the mechanical strength of CNS tissue. The hydrogels were found to have shear thinning properties desirable for injectable formulations and they also maintained higher Schwann cell viability during injection compared to saline controls. Usingin vitroco-culture experiments, we found that the genipin-crosslinked hydrogels also protected Schwann cells from astrocyte-mediated inflammation.Significance. Injectable hydrogels developed using delipidated and decellularized porcine peripheral nerves are a potential clinically relevant solution to deliver Schwann cells, and possibly other therapeutic cells, at the SCI site by maintaining higher cellular viability and increasing therapeutic efficacy for SCI treatment.

A 2-step remote TUNEL approach for sperm DNA fragmentation assessment. Analysis in donors and patients.

OBJECTIVES: Infertility is a disease of the male or female reproductive systems. Male reproductive workup is based on routine semen analysis, although of limited value. The 2021 WHO Manual incorporated Sperm DNA Fragmentation (SDF) assessment, and highlighted the need for individual laboratories to define suitable thresholds. This study aimed to present an alternative to address this issue, determine an SDF cut-off value with fertile donors, and characterize SDF in a patient cohort and their relationship with semen parameters. STUDY DESIGN: A service unit was established to remotely perform TUNEL assay in a 2 step-process. Semen samples were received at andrology laboratories, subjected to routine semen analysis (WHO, 2010), partially processed and transported to the service unit for SDF evaluation. Using this setting, studies were done in fertile donors (n = 15) to define the cut-off value, and in men undergoing infertility workup (n = 318). RESULTS: A cut-off value of 9.17 % was determined with the fertile donor cohort. With this cut-off, a 64.46 % abnormal SDF incidence was determined in the patient cohort. SDF negatively correlated with sperm number, vitality and motility, and positively with abnormal morphology and male age (P < 0.05). TUNEL-positive cases depicted lower sperm quality and higher male age (P < 0.05). A similar abnormal SDF incidence was determined among patients with semen abnormalities. Asthenozoospermic and ≥40 years patient samples depicted higher (P < 0.05) SDF than those of the general population. SDF incidence was also high in normozoospermic patients. CONCLUSIONS: Using a 2-step remote approach with a standardized procedure and an SDF cut-off value established with fertile donors, high SDF incidence in semen samples depicting normal and abnormal quality were identified in men consulting for infertility, highlighting the relevance of its evaluation as part of the male fertility workup.

Vectorial Image Representation for Image Classification.

This paper proposes the transformation S→C→, where S is a digital gray-level image and C→ is a vector expressed through the textural space. The proposed transformation is denominated Vectorial Image Representation on the Texture Space (VIR-TS), given that the digital image S is represented by the textural vector C→. This vector C→ contains all of the local texture characteristics in the image of interest, and the texture unit T→ entertains a vectorial character, since it is defined through the resolution of a homogeneous equation system. For the application of this transformation, a new classifier for multiple classes is proposed in the texture space, where the vector C→ is employed as a characteristics vector. To verify its efficiency, it was experimentally deployed for the recognition of digital images of tree barks, obtaining an effective performance. In these experiments, the parametric value λ employed to solve the homogeneous equation system does not affect the results of the image classification. The VIR-TS transform possesses potential applications in specific tasks, such as locating missing persons, and the analysis and classification of diagnostic and medical images.

Preparing for the Next Pandemic: Increased Expression of Interferon-Stimulated Genes After Local Administration of Nasalferon or HeberNasvac.

HeberNasvac, a therapeutic vaccine for chronic hepatitis B, is able to safely stimulate multiple Toll-like receptors, increasing antigen presentation in vitro and in a phase II clinical trial (Profira) in elderly volunteers who were household contacts of respiratory infection patients. Thus, a new indication as a postexposure prophylaxis or early therapy for respiratory infections has been proposed. In this study, we evaluated the expression of several interferon-stimulated genes (ISGs) after mucosal administration of HeberNasvac and compared this effect with the nasal delivery of interferon alpha 2b (Nasalferon). Molecular studies of blood samples of 50 subjects from the Profira clinical trial who were locally treated with HeberNasvac or Nasalferon and concurrent untreated individuals were compared based on their relative mRNA expression of OAS1, ISG15, ISG20, STAT1, STAT3, and DRB1-HLA II genes. In most cases, the gene expression induced by HeberNasvac was similar in profile and intensity to the expression induced by Nasalferon and significantly superior to that observed in untreated controls. The immune stimulatory effect of HeberNasvac on ISGs paved the way for its future use as an innate immunity stimulator in elderly persons and immunocompromised subjects or as part of Mambisa, a nasal vaccine to prevent severe acute respiratory syndrome coronavirus 2 infection.

Human skeletal muscle tissue chip autonomous payload reveals changes in fiber type and metabolic gene expression due to spaceflight.

Microphysiological systems provide the opportunity to model accelerated changes at the human tissue level in the extreme space environment. Spaceflight-induced muscle atrophy experienced by astronauts shares similar physiological changes to muscle wasting in older adults, known as sarcopenia. These shared attributes provide a rationale for investigating molecular changes in muscle cells exposed to spaceflight that may mimic the underlying pathophysiology of sarcopenia. We report the results from three-dimensional myobundles derived from muscle biopsies from young and older adults, integrated into an autonomous CubeLab™, and flown to the International Space Station (ISS) aboard SpaceX CRS-21 as part of the NIH/NASA funded Tissue Chips in Space program. Global transcriptomic RNA-Seq analyses comparing the myobundles in space and on the ground revealed downregulation of shared transcripts related to myoblast proliferation and muscle differentiation. The analyses also revealed downregulated differentially expressed gene pathways related to muscle metabolism unique to myobundles derived from the older cohort exposed to the space environment compared to ground controls. Gene classes related to inflammatory pathways were downregulated in flight samples cultured from the younger cohort compared to ground controls. Our muscle tissue chip platform provides an approach to studying the cell autonomous effects of spaceflight on muscle cell biology that may not be appreciated on the whole organ or organism level and sets the stage for continued data collection from muscle tissue chip experimentation in microgravity. We also report on the challenges and opportunities for conducting autonomous tissue-on-chip CubeLabTM payloads on the ISS.

Validation of Human Skeletal Muscle Tissue Chip Autonomous Platform to Model Age-Related Muscle Wasting in Microgravity.

Microgravity-induced muscle atrophy experienced by astronauts shares similar physiological changes to muscle wasting experienced by older adults, known as sarcopenia. These shared attributes provide a rationale for investigating microgravity-induced molecular changes in human bioengineered muscle cells that may also mimic the progressive underlying pathophysiology of sarcopenia. Here, we report the results of an experiment that incorporated three-dimensional myobundles derived from muscle biopsies from young and older adults, that were integrated into an autonomous CubeLabâ"¢, and flown to the International Space Station (ISS) aboard SpaceX CRS-21 in December 2020 as part of the NIH/NASA funded Tissue Chips in Space program. Global transcriptomic RNA-Seq analysis comparing the myobundles in space and on the ground revealed downregulation of shared transcripts related to myoblast proliferation and muscle differentiation for those in space. The analysis also revealed differentially expressed gene pathways related to muscle metabolism unique to myobundles derived from the older cohort exposed to the space environment compared to ground controls. Gene classes related to inflammatory pathways were uniquely modulated in flight samples cultured from the younger cohort compared to ground controls. Our muscle tissue chip platform provides a novel approach to studying the cell autonomous effects of microgravity on muscle cell biology that may not be appreciated on the whole organ or organism level and sets the stage for continued data collection from muscle tissue chip experimentation in microgravity. Thus, we also report on the challenges and opportunities for conducting autonomous tissue-on-chip CubeLab TM payloads on the ISS.

Microphysiological system for studying contractile differences in young, active, and old, sedentary adult derived skeletal muscle cells.

Microphysiological systems (MPS), also referred to as tissue chips, incorporating 3D skeletal myobundles are a novel approach for physiological and pharmacological studies to uncover new medical treatments for sarcopenia. We characterize a MPS in which engineered skeletal muscle myobundles derived from donor-specific satellite cells that model aged phenotypes are encapsulated in a perfused tissue chip platform containing platinum electrodes. Our myobundles were derived from CD56+ myogenic cells obtained via percutaneous biopsy of the vastus lateralis from adults phenotyped by age and physical activity. Following 17 days differentiation including 5 days of a 3 V, 2 Hz electrical stimulation regime, the myobundles exhibited fused myotube alignment and upregulation of myogenic, myofiber assembly, signaling and contractile genes as demonstrated by gene array profiling and localization of key components of the sarcomere. Our results demonstrate that myobundles derived from the young, active (YA) group showed high intensity immunofluorescent staining of α-actinin proteins and responded to electrical stimuli with a ~1 µm displacement magnitude compared with non-stimulated myobundles. Myobundles derived from older sedentary group (OS) did not display a synchronous contraction response. Hypertrophic potential is increased in YA-derived myobundles in response to stimulation as shown by upregulation of insulin growth factor (IGF-1), α-actinin (ACTN3, ACTA1) and fast twitch troponin protein (TNNI2) compared with OS-derived myobundles. Our MPS mimics disease states of muscle decline and thus provides an aged system and experimental platform to investigate electrical stimulation mimicking exercise regimes and may be adapted to long duration studies of compound efficacy and toxicity for therapeutic evaluation against sarcopenia.

Effect of 16% Carbamide Peroxide and Activated-Charcoal-Based Whitening Toothpaste on Enamel Surface Roughness in Bovine Teeth: An In Vitro Study.

Background: Activated charcoal is a nanocrystalline form of carbon with a large specific surface area and high porosity in the nanometer range, having consequently the capacity to absorb pigments, chromophores, and stains responsible for tooth color change, while carbamide peroxide is unstable and breaks down immediately upon contact with tissue and saliva, first dissociating into hydrogen peroxide and urea and subsequently into oxygen, water, and carbon dioxide. Therefore, the aim of the present study was to assess the effect of 16% carbamide peroxide and activated-charcoal-based whitening toothpaste on enamel surface roughness in bovine teeth. Materials and Methods: The present experimental in vitro, longitudinal, and prospective study consisted of 60 teeth randomly distributed in six groups: A: artificial saliva, B: conventional toothpaste (Colgate Maximum Protection), C: whitening toothpaste with activated charcoal (Oral-B 3D White Mineral Clear), D: 16% carbamide peroxide (Whiteness Perfect 16%), E: 16% carbamide peroxide plus conventional toothpaste (Whiteness Perfect 16% plus Colgate Maximum Protection), and F: 16% carbamide peroxide plus whitening toothpaste with activated charcoal (Whiteness Perfect 16% plus Oral-B 3D White Mineral Clear). Surface roughness was assessed with a digital roughness meter before and after each treatment. For the statistical analysis, Student's t test for related samples was used, in addition to the ANOVA test for one intergroup factor, considering a significance level of p < 0.05. Results: The surface roughness variation of bovine tooth enamel, before and after application of bleaching agent, was higher in groups of whitening toothpaste with activated charcoal (0.200 µm, Confidence Interval (CI): 0.105; 0.296 µm) and 16% carbamide peroxide plus whitening toothpaste with activated charcoal (0.201 µm, (CI): 0.092; 0.309 µm). In addition, bovine teeth treated with conventional toothpaste (p = 0.041), whitening toothpaste with activated charcoal (p = 0.001), and 16% carbamide peroxide plus whitening toothpaste with activated charcoal (p = 0.002) significantly increased their surface roughness values. On the other hand, significant differences were observed when comparing the variation in surface roughness between the application of artificial saliva (control) and the whitening toothpaste with activated charcoal (p = 0.031), and the 16% carbamide peroxide plus whitening toothpaste with activated charcoal (p = 0.030). Conclusion: The use of whitening toothpaste with activated charcoal and in combination with 16% carbamide peroxide significantly increased enamel surface roughness in bovine teeth.

Development of a magnetically aligned regenerative tissue-engineered electronic nerve interface for peripheral nerve applications.

Peripheral nerve injuries can be debilitating to motor and sensory function, with severe cases often resulting in complete limb amputation. Over the past two decades, prosthetic limb technology has rapidly advanced to provide users with crude motor control of up to 20° of freedom; however, the nerve-interfacing technology required to provide high movement selectivity has not progressed at the same rate. The work presented here focuses on the development of a magnetically aligned regenerative tissue-engineered electronic nerve interface (MARTEENI) that combines polyimide "threads" encapsulated within a magnetically aligned hydrogel scaffold. The technology exploits tissue-engineered strategies to address concerns over traditional peripheral nerve interfaces including poor axonal sampling through the nerve and rigid substrates. A magnetically templated hydrogel is used to physically support the polyimide threads while also promoting regeneration in close proximity to the electrode sites on the polyimide. This work demonstrates the utility of magnetic templating for use in tuning the mechanical properties of hydrogel scaffolds to match the stiffness of native nerve tissue while providing an aligned substrate for Schwann cell migration in vitro. MARTEENI devices were fabricated and implanted within a 5-mm-long rat sciatic-nerve transection model to assess regeneration at 6 and 12 weeks. MARTEENI devices do not disrupt tissue remodeling and show axon densities equivalent to fresh tissue controls around the polyimide substrates. Devices are observed to have attenuated foreign-body responses around the polyimide threads. It is expected that future studies with functional MARTEENI devices will be able to record and stimulate single axons with high selectivity and low stimulation regimes.

One­carbon metabolism factor MTHFR variant is associated with saccade latency in Spinocerebellar Ataxia type 2.

BACKGROUND: Spinocerebellar ataxia type 2 (SCA2) is a neurodegenerative disorder due to a CAG-repeat expansion. This work is intended to identify modifiers of the clinical phenotype in SCA2, following up on recent genome-wide association analyses that demonstrated the prominent role of DNA-damage repair and methylation for the severity and progression of polyglutamine diseases. In particular, we assessed the impact of MTHFR as rate-limiting enzyme in DNA methylation pathways, which modulates cerebellar neurotransmission and motor neuron atrophy. METHODS: A sample of 166 Cuban SCA2 patients and of 130 healthy subjects from the same geographical and ethnic background was selected. The ATXN2 CAG repeat length was determined by PCR followed by polyacrylamide gel electrophoresis. Two amino acid substitutions known to decrease the enzyme activity of MTHFR, encoded by C677T and A1298C polymorphisms, were assessed by PCR/RFLP. RESULTS: No significant differences were observed for C677T or A1298C alleles or genotype frequencies between cases and controls, confirming that disease risk in SCA2 does not depend on MTHFR activity. However, MTHFR A1298C genotypes showed a significant association with saccade latency. CONCLUSIONS: \MTHFR A1298C polymorphism is associated with saccade latency in SCA2 patients, but not with disease risk, age at onset or maximal saccade velocity. These results provide evidence that folate-mediated one­carbon metabolism might be important in the physiopathology of SCA2.

Screening of nutritional risk: assessment of predictive variables of nutritional risk in hospitalized patients in a second-level care center in Mexico / Tamizaje de riesgo nutricional: evaluación de variables predictivas de riesgo nutricional en pacientes hospitalizados en un centro de atención de segundo nivel en México

Introduction: worldwide, hospital malnutrition constitutes an important issue of morbidity and mortality. Although the prevalence of malnutrition has been calculated as between 7% and 27% in hospitalized patients, its real prevalence remains unknown or underestimated because of the different criteria for its identification and diagnosis. The aim of this study was to determine the prevalence of nutritional risk in a cohort of hospitalized patients and to identify the significance of the predictors associated with nutritional risk. Methods: the evaluation of the presence of nutritional risk was carried out in 247 individuals hospitalized at the second-level care institution Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, in Mexico. Nutritional screening was evaluated during the first 24 hours of stay with the NRS 2002. The weighing of associated variables with nutritional risk was calculated statistically using the software Sigma Plot v11. Results: forty-two percent of patients were at risk of malnutrition. Significant associations between nutritional risk and a reduction in food ingestion (during the last week), the illness severity of the patient, as well as age and sex (p < 0.05), were observed. A reduction in food ingestion during the previous week increased the likelihood of having nutritional risk 6.67 times more (95% CI: 3.4-13.2; p < 0.001) in the studied population. Conclusion: the risk of malnutrition in hospitalized patients at ISSSTE-Zacatecas, Hospital General Nº 26 is frequent (42%). Therefore, early detection of nutritional risk is important to offer for proper nutritional intervention with the objective of decreasing the associated morbidity and mortality

Introducción: la desnutrición hospitalaria constituye un problema de morbimortalidad en todo el mundo. Aunque la prevalencia de malnutrición se ha calculado entre el 7% y el 27% en pacientes hospitalizados, su prevalencia real sigue siendo desconocida o subestimada debido a los diferentes criterios para su identificación y diagnóstico. El objetivo de este estudio fue determinar la prevalencia del riesgo nutricional mediante la herramienta Nutritional Risk Screening 2002 (NRS-2002) en pacientes hospitalizados y ponderar los factores predictivos asociados con el riesgo nutricional. Métodos: la evaluación de la presencia de riesgo nutricional se realizó en 247 hospitalizados en el hospital general de segundo nivel Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, en México. La evaluación nutricional se realizó durante las primeras 24 horas de estadía mediante la herramienta NRS 2002. El análisis de datos se llevó a cabo mediante el software Sigma Plot v11. Resultados: el 42% de los pacientes presentaron riesgo de desnutrición. Después de la corrección por covariables, se encontraron asociaciones significativas entre el riesgo nutricional y una reducción de la ingesta de alimentos (durante la última semana), la gravedad de la enfermedad del paciente, la edad y el sexo (p < 0,05). Entre la población estudiada, la reducción de la ingesta durante la última semana aumentó 6,67 veces la probabilidad de presentar riesgo nutricional (IC 95%: 3,4-13,2; p < 0,001). Conclusión: el riesgo de desnutrición en pacientes hospitalizados en el Hospital General Nº 26 Zacatecas-ISSSTE es frecuente (42%), por lo que es importante realizar una detección temprana para ofrecer una intervención nutricional adecuada y, con ello, disminuir la morbimortalidad asociada

Screening of nutritional risk: assessment of predictive variables of nutritional risk in hospitalized patients in a second-level care center in Mexico.

INTRODUCTION: Introduction: worldwide, hospital malnutrition constitutes an important issue of morbidity and mortality. Although the prevalence of malnutrition has been calculated as between 7% and 27% in hospitalized patients, its real prevalence remains unknown or underestimated because of the different criteria for its identification and diagnosis. The aim of this study was to determine the prevalence of nutritional risk in a cohort of hospitalized patients and to identify the significance of the predictors associated with nutritional risk. Methods: the evaluation of the presence of nutritional risk was carried out in 247 individuals hospitalized at the second-level care institution Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, in Mexico. Nutritional screening was evaluated during the first 24 hours of stay with the NRS 2002. The weighing of associated variables with nutritional risk was calculated statistically using the software Sigma Plot v11. Results: forty-two percent of patients were at risk of malnutrition. Significant associations between nutritional risk and a reduction in food ingestion (during the last week), the illness severity of the patient, as well as age and sex (p < 0.05), were observed. A reduction in food ingestion during the previous week increased the likelihood of having nutritional risk 6.67 times more (95% CI: 3.4-13.2; p < 0.001) in the studied population. Conclusion: the risk of malnutrition in hospitalized patients at ISSSTE-Zacatecas, Hospital General Nº 26 is frequent (42%). Therefore, early detection of nutritional risk is important to offer for proper nutritional intervention with the objective of decreasing the associated morbidity and mortality.

INTRODUCCIÓN: Introducción: la desnutrición hospitalaria constituye un problema de morbimortalidad en todo el mundo. Aunque la prevalencia de malnutrición se ha calculado entre el 7% y el 27% en pacientes hospitalizados, su prevalencia real sigue siendo desconocida o subestimada debido a los diferentes criterios para su identificación y diagnóstico. El objetivo de este estudio fue determinar la prevalencia del riesgo nutricional mediante la herramienta Nutritional Risk Screening 2002 (NRS-2002) en pacientes hospitalizados y ponderar los factores predictivos asociados con el riesgo nutricional. Métodos: la evaluación de la presencia de riesgo nutricional se realizó en 247 hospitalizados en el hospital general de segundo nivel Instituto de Seguridad y Servicios Sociales para los Trabajadores del Estado (ISSSTE) Zacatecas, Hospital General Nº 26, en México. La evaluación nutricional se realizó durante las primeras 24 horas de estadía mediante la herramienta NRS 2002. El análisis de datos se llevó a cabo mediante el software Sigma Plot v11. Resultados: el 42% de los pacientes presentaron riesgo de desnutrición. Después de la corrección por covariables, se encontraron asociaciones significativas entre el riesgo nutricional y una reducción de la ingesta de alimentos (durante la última semana), la gravedad de la enfermedad del paciente, la edad y el sexo (p < 0,05). Entre la población estudiada, la reducción de la ingesta durante la última semana aumentó 6,67 veces la probabilidad de presentar riesgo nutricional (IC 95%: 3,4-13,2; p < 0,001). Conclusión: el riesgo de desnutrición en pacientes hospitalizados en el Hospital General Nº 26 Zacatecas-ISSSTE es frecuente (42%), por lo que es importante realizar una detección temprana para ofrecer una intervención nutricional adecuada y, con ello, disminuir la morbimortalidad asociada.

Resorbable plating system stabilizes tissue-engineered intervertebral discs implanted ex vivo in canine cervical spines.

Total disc replacement using tissue-engineered intervertebral discs (TE-IVDs) may offer a biological alternative to treat radiculopathy caused by disc degeneration. A composite TE-IVD was previously developed and evaluated in rat tail and beagle cervical spine models in vivo. Although cell viability and tissue integration into host tissue were promising, significant implant displacement occurred at multiple spinal levels. The goal of the present study was to assess the effects of a resorbable plating system on the stiffness of motion segments and stability of tissue-engineered implants subjected to axial compression. Canine motion segments from levels C2/C3 to C5/C6 were assessed as intact (CTRL), after discectomy (Dx), with an implanted TE-IVD only (PLATE-), and with a TE-IVD combined with an attached resorbable plate (PLATE+). Segments under PLATE+ conditions fully restored separation between endplates and showed significantly higher compressive stiffness than segments under PLATE- conditions. Plated segments partially restored more than 25% of the CTRL motion segment stiffness. Plate attachment also prevented implant extrusion from the disc space at 50% compressive strain, and this effect was more significant in segments from levels C3/C4 when compared to segments from level C5/C6. These results suggest that stabilization of motion segments via resorbable plating assists TE-IVD retention in the disc space while allowing the opportunity for implants to fully integrate into the host tissue and achieve optimal restoration of spine biomechanics.

Ovarian and Breast Cancer Migration Dynamics on Laminin and Fibronectin Bidirectional Gradient Fibers Fabricated via Multiphoton Excited Photochemistry.

INTRODUCTION: Migration mis-regulation is a hallmark of cancer, and remains an important problem in cancer biology. We postulate the needs for better in vitro models to understand the details of cell-matrix interactions. Here, we utilized multiphoton excited (MPE) photochemistry to fabricate models to systematically study migration dynamics operative in breast and ovarian cancer. Gradients are a convenient means to modulate concentration and also have been implicated in metastases. METHODS: We specifically pattern sub-micron structured gradients from laminin and fibronectin whose up-regulation is associated with increased metastasis and poor prognosis. We developed a new continuous linear bi-directional gradient design, permitting exploration of the underlying cell-matrix interactions of migration, including speed, directness, and f-actin cytoskeleton alignment as a function of concentration. These new models provide both contact guidance and ECM binding cues, and provide a more relevant environment than possible with existing technologies such as flow chambers or 2D printed surfaces. RESULTS: We found an overall increase in these processes with increasing concentration on both laminin and fibronectin gradients for a series of ovarian and breast cancer lines. Moreover, directness was higher for more metastatic cells, indicating that epithelial or mesenchymal state of the cell type governs the dynamics. However, the specifics of the speed and directedness depend on both the cell type and protein, thus we found that we must consider these processes collectively to obtain a self-consistent picture of the migration. For this purpose, we performed a linear discriminate analysis (LDA) and successfully classified the different cell types on the two protein gradients without molecular biology analysis. CONCLUSIONS: The bi-gradient structures are versatile tools to performing detailed studies of cell migration, specifically haptotxis. We further suggest the can be used in assessing efficacy of drug treatments targeted at specific matrix components.

Total disc replacement using tissue-engineered intervertebral discs in the canine cervical spine.

The most common reason that adults in the United States see their physician is lower back or neck pain secondary to degenerative disc disease. To date, approaches to treat degenerative disc disease are confined to purely mechanical devices designed to either eliminate or enable flexibility of the diseased motion segment. Tissue engineered intervertebral discs (TE-IVDs) have been proposed as an alternative approach and have shown promise in replacing native IVD in the rodent tail spine. Here we demonstrate the efficacy of our TE-IVDs in the canine cervical spine. TE-IVD components were constructed using adult canine annulus fibrosis and nucleus pulposus cells seeded into collagen and alginate hydrogels, respectively. Seeded gels were formed into a single disc unit using molds designed from the geometry of the canine spine. Skeletally mature beagles underwent discectomy with whole IVD resection at levels between C3/4 and C6/7, and were then divided into two groups that received only discectomy or discectomy followed by implantation of TE-IVD. Stably implanted TE-IVDs demonstrated significant retention of disc height and physiological hydration compared to discectomy control. Both 4-week and 16-week histological assessments demonstrated chondrocytic cells surrounded by proteoglycan-rich matrices in the NP and by fibrocartilaginous matrices in the AF portions of implanted TE-IVDs. Integration into host tissue was confirmed over 16 weeks without any signs of immune reaction. Despite the significant biomechanical demands of the beagle cervical spine, our stably implanted TE-IVDs maintained their position, structure and hydration as well as disc height over 16 weeks in vivo.

Characteristics of bacterial acute diarrhea among women.

OBJECTIVE: To describe the clinical and microbiological characteristics of women presenting with acute gastroenteritis due to infection with Salmonella spp. or Shigella spp. METHODS: A cross-sectional retrospective study was conducted using data for patients with specimens positive for Shigella spp. and Salmonella spp. attending two major women's hospitals in Porto Alegre, Brazil, between January 2003 and July 2014. Women were included if they had symptoms compatible with bacterial acute diarrhea. Isolates were evaluated to determine antimicrobial susceptibility and patient clinical profile. RESULTS: Among 45 eligible patients, Salmonella spp. was identified in 32 (71%) and Shigella spp. in 13 (29%). The highest antimicrobial sensitivity rates were observed for ciprofloxacin and ceftriaxone (n=44, 98% for both) whereas the greatest resistance rate was seen for ampicillin (n=20, 44%). Seven (16%) of the women were pregnant. CONCLUSION: Ciprofloxacin, ceftriaxone, and sulfamethoxazole/trimethoprim are suitable choices for the treatment of bacterial acute diarrhea. However, the maternal and fetal safety profile of prescribed medications should be considered when treating pregnant patients.

Association of leishmaniasis with TNF alpha promoter and SLC11A1 gene polymorphisms in patients of two endemic areas in Mexico.

Some Single Nucleotide Polymorphisms (SNPs) of interleukins and other modulatory molecules of the immune response play an important role in susceptibility to infectious diseases, particularly those involving intracellular parasites. In this study, we evaluated allele, genotype and haplotype associations of two SNPs of the TNF-α promoter and seven of the SLC11A1 gene in 79 patients with localized cutaneous leishmaniasis (CL) and 15 with visceral leishmaniasis (VL), compared with 127 and 89 locality paired controls, respectively, from two endemic areas of Chiapas State, Mexico. None of the TNF-α alleles and genotypes was associated either to CL or to VL. Alleles rs2276631-C (P = 0.02; OR [95%CI] = 2.11 [1.16-3.86]) and rs2279015-G (P = 0.005; OR [95%CI] = 2.42 [1.33-4.41]) of SLC11A1, were associated with susceptibility to VL, whereas genotypes rs2276631 C/C (P = 0.003; OR [95%CI] = 2.65 [1.41-5.00]) and rs2279015 G/G (P = 0.018; OR [95%CI] = 2.05 [1.15-3.64]) were significantly increased in CL and VL patients, respectively. Complete haplotypes involved in susceptibility were CGCCGDins with VL and CGCCADins with CL. CGCCA was the minimal susceptibility haplotype for CL and CCG for VL. Our data suggest that SLC11A1 gene polymorphisms might have a relevant role in the pathology of leishmaniasis, directing towards susceptibility outcome of this disease in residents of an endemic area.

Trifluoroacetic acid as excipient destabilizes melittin causing the selective aggregation of melittin within the centrin-melittin-trifluoroacetic acid complex.

Trifluoroacetic acid (TFA) may be the cause of the bottleneck in high resolution structure determination for protein-peptide complexes. Fragment based drug design often involves the use of synthetic peptides which contain TFA (excipient). Our goal was to explore the effects of this excipient on a model complex: centrin-melittin-TFA. We performed Fourier transform infrared, two-dimensional infrared correlation spectroscopies and spectral simulations to analyze the amide I'/I'* band for the components and the ternary complex. Melittin (MLT) was observed to have increased helicity upon its interaction with centrin, followed by the thermally induced aggregation of MLT within the ternary complex in the TFA presence.

Gas Permeation Properties of Soluble Aromatic Polyimides Based on 4-Fluoro-4,4'-Diaminotriphenylmethane.

A series of new organic polyimides were synthesized from 4-fluoro-4'4"-diaminotriphenylmethane and four different aromatic dianhydrides through a one-step, high-temperature, direct polycondensation in m-cresol at 180-200 °C, resulting in the formation of high-molecular-weight polyimides (inherent viscosities ~ 1.0-1.3 dL/g). All the resulting polyimides exhibited good thermal stability with initial decomposition temperatures above 434 °C, glass-transition temperatures between 285 and 316 °C, and good solubility in polar aprotic solvents. Wide-angle X-ray scattering data indicated that the polyimides were amorphous. Dense membranes were prepared by solution casting and solvent evaporation to evaluate their gas transport properties (permeability, diffusivity, and solubility coefficients) toward pure hydrogen, helium, oxygen, nitrogen, methane, and carbon dioxide gases. In general, the gas permeability was increased as both the fractional free volume and d-spacing were also increased. A good combination of permeability and selectivity was promoted efficiently by the bulky hexafluoroisopropylidene and 4-fluoro-phenyl groups introduced into the polyimides. The results indicate that the gas transport properties of these films depend on both the structure of the anhydride moiety, which controls the intrinsic intramolecular rigidity, and the 4-fluoro-phenyl pendant group, which disrupts the intermolecular packing.