A platform to understand amyotrophic lateral sclerosis (ALS) and extend human motor neurons longevity / Plataforma para empezar a comprender la esclerosis lateral amiotrófica (ELA) y extender la longevidad de neuronas motoras humanas

In vitro modeling of neurodegenerative diseases is now possible by using patient-derived induced pluripotent stem cells (iPS). Through them, it is nowadays conceivable to obtain human neurons and glia, and study diseases cellular and molecular mechanisms, an attribute that was previously unavailable to any human condition. Amyotrophic lateral sclerosis (ALS) is one of the diseases that has gained a rapid advance with iPS technology. By differentiating motor neurons from iPS cells of ALS- patients, we are studying the mechanisms underlying ALS- disease onset and progression. Here, we introduce a cellular platform to help maintain longevity of ALS iPS-motor neurons, a cellular feature relevant for most late-onset human diseases. Long term cultures of patient-derived iPS cells might prove to be critical for the development of personalized-drugs.

Actualmente es posible modelar in vitro enfermedades neurodegenerativas humanas mediante el uso de células madre pluripotentes inducidas (iPS) derivadas del paciente. A través de ellas, es hoy concebible obtener neuronas y glía humanas, y estudiar mecanismos celulares y moleculares de enfermedades, un atributo que anteriormente no era posible para ninguna condición humana. La esclerosis lateral amiotrófica (ELA) es una de las enfermedades que se ha beneficiado con la tecnología de iPS. Al diferenciar neuronas motoras de células iPS obtenidas de pacientes con ELA, hemos iniciado estudios sobre los mecanismos que subyacen a la aparición y progresión de la enfermedad. Aquí, presentamos el desarrollo de una plataforma celular que permite extender la longevidad de las neuronas motoras derivadas de iPS, una característica relevante para la mayoría de las enfermedades humanas de inicio tardío. Los cultivos a largo plazo de células iPS provenientes de pacientes pueden ser determinantes en el desarrollo de terapias asociadas a la medicina de precisión.

Social cohesion, condom use, and sexually transmitted infections among female sex workers living with HIV in the Dominican Republic.

This cross-sectional study examines the relationship between social cohesion with consistent condom use (CCU) and sexually transmitted infections (STIs) among the Abriendo Puertas (Opening Doors) cohort of female sex workers (FSWs) living with human immunodeficiency virus (HIV) in the Dominican Republic (n = 228). Using data from the follow-up survey of the cohort, we conducted multivariate logistic regression to explore these dynamics. Social cohesion was significantly associated with CCU between FSWs living with HIV and their clients in the last month (adjusted odds ratio [AOR] = 1.65, 95% confidence interval [CI]: 1.11-2.45) and STI prevalence among FSWs (AOR: 3.76, CI: 1.159-12.162). Social cohesion was not associated with CCU between FSWs living with HIV and their steady partners. However, both illicit drug use in the past six months (AOR = 0.11, CI: 0.023-0.57) and pregnancy intentions (AOR = 0.11; CI: 0.02-0.42) were significantly associated with CCU with steady partners. Findings highlight the differential role of social cohesion on condom use outcomes between FSWs living with HIV and their paying clients versus steady partners. Research on the pathways via which cohesion influences condom use among sex workers and their clients is merited, as is research regarding the role of drug use and pregnancy intentions on condom use with steady partners.

Clinico-pathological features of the intradiaphragmatic bronchogenic cysts: report of a case and review of the literature.

Bronchogenic cysts represent congenital malformations deriving from an abnormal development of the primitive foregut during embryogenesis. These lesions are rarely found and they are most frequently localized in the mediastinum, or in lung parenchyma. Intramuscular localization is extremely rare, especially within the diaphragm. We report a case of a 54 year old man showing a large lobulated cystic lesion in the left hemidiaphragm. Complete surgery was performed and histological diagnosis of intradiaphragmatic bronchogenic cyst was made during surgery and confirmed after a total section analysis. Moreover we reviewed the other cases published in the English literature, including clinical, surgical and pathological data.

Hamate fractures.

OBJECTIVE: The purpose of this paper is to present our experience in the treatment of the fractures of the hamate and to make a review of the literature on this topic. MATERIAL AND METHOD: We retrospectively reviewed 10 patients treated in our clinic between 2005-2012 suffering from fractures of the hamate. Six cases were fractures of the body and four were fractures of the hamate. Five cases were of associated injuries. Diagnostic delay ranged from 30 days to 2 years. Patient follow-up ranged from 1 to 10 years. Patient satisfaction was evaluated using the DASH questionnaire. Five patients with a fracture of the body underwent surgery, and one was treated conservatively. Two patients with fracture of the hook of the hamate were treated with immobilization, and two more patients had the fragment removed. RESULTS: The grip strength and the digital clip were reduced in 2 cases. Flexion and extension of the wrist was limited in 3 cases. The mobility of the fingers was normal in all the cases, except in one. The results obtained from the DASH questionnaire were normal in all the cases, except in one case of fracture of the hamate, and in two cases of fracture of the body. CONCLUSIONS: The surgical treatment should reduce the dislocation and stabilize the injuries with osteosynthesis. The fractures of the hamate are usually diagnosed late, and the most recommended treatment is removal of the fragment, although it cannot be deduced from this study.

A comparison between radiochromic EBT2 film model and its predecessor EBT film model.

The manufacturer has introduced the new EBT2 film model so as to improve its predecessor, the EBT radiochromic film model. According to the manufacturer, some of its main advantages include a higher tolerance to light exposure and it can correct non-uniformity of the active layer thickness using a marker dye. However, the equivalence in uniformity between both models was questioned by some authors, and the asymmetrical configuration of layers of the EBT2 film model produces a new dependence on the film side being scanned (front and back orientation). In this study, the EBT2 radiochromic film model was compared with the EBT model and the new marker dye feature was assessed. We also compared this correction method with a pre-irradiated pixel value correction method. An Epson Expression 10000XL scanner in transmission mode was used to scan the films and the red channel response was analyzed. We confirmed the lower-measured signal dependence on the visible light exposure of the EBT2 film model. Differences in pixel values remained below 0.5% for a minimum of 15 days. In regard to the uniformity, similar results for EBT2 and EBT film models were obtained; in both cases inhomogeneity was found to be less than 1%, in relative pixel value from the mean. However, we found that the signal-to-noise ratio was reduced for low doses by 37% for old EBT2 batch and by 21% for new EBT2 batch compared to signal-to-noise ratio for EBT. The EBT2 film model's pixel value difference for the front and back orientation reached up to 1.0% in the red channel. Our results did not show a clear advantage between to use a pre-irradiated pixel value correction and to use the manufacturer's correction.

Relative and absolute stability in perceived parenting behaviour: a longitudinal study with children and adolescents.

Patterns of relative and absolute stability in parental behaviour with children and adolescents are reported. The sample comprised 523 youth (58.7% girls). Data were collected at three time periods: T1 (M age = 11.1 yr.), T2 (M age = 12.2 yr.), and T3 (M age = 13.2 yr.), each separated by one year. According to children's reports, relative consistency was moderate in both mothers and fathers, particularly as regards communication and strict control. In contrast, as children got older, parental rearing practices related to strict control and hostility decreased. There was a similarity between fathers and mothers in terms of relative and absolute stability. Relative stability was affected by the child's sex, the parenting variable, and the time period; however, the patterns of absolute stability reveal no differences by sex.

Bipolar radiofrequency in the human meniscus. Comparative study between patients younger and older than 40 years of age.

PURPOSE: To morphohistologically evaluate the effect of four increasing intensities of bipolar radiofrequency (RF) on the human meniscus and to compare the changes seen in the menisci from patients younger and older than 40 years old. METHODS: Thirty fresh menisci were divided in two groups. Group 1: 12 menisci from patients younger than 40 y.o.; Group 2: 18 menisci from patients older than 40 y.o. Groups 1 and 2 were divided in four zones and subjected in vitro, for 3 s, to four intensities of bipolar RF energy. The samples were studied macroscopically, and microscopically. RESULTS: Thermal changes were present between 0 and 4000 microm from the meniscal surface, with an average of 1699 microm (S.D. 740), and was significantly higher on the ablation than the coagulation group (p<0.001). We found a significant difference between the depth of thermal changes in the menisci from patients younger and older than 40 years old at medium intensities of RF energy (p=0.038 and p=0.044). CONCLUSIONS: Although bipolar RF can cause deep thermal changes (up to 4000 microm) on the human meniscus, this effect depends on the magnitude of the energy applied. When comparing the effect between younger and older patients, the changes were deeper in the older group when RF was applied at medium intensities. CLINICAL RELEVANCE: Based on our findings, we recommend to exert precaution when applying RF energy at medium intensities in the degenerative meniscus, due to a deeper thermal effect in this group.

Regulation of glycinergic and GABAergic synaptogenesis by brain-derived neurotrophic factor in developing spinal neurons.

Brain-derived neurotrophic factor (BDNF) effects on the establishment of glycinergic and GABAergic transmissions in mouse spinal neurons were examined using combined electrophysiological and calcium imaging techniques. BDNF (10 ng/ml) caused a significant acceleration in the onset of synaptogenesis without large effects on the survival of these neurons. Amplitude and frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) and miniature inhibitory postsynaptic currents (mIPSCs) associated to activation of glycine and GABA(A) receptors were augmented in neurons cultured with BDNF. The neurotrophin effect was blocked by long term tetrodotoxin (TTX) addition suggesting a dependence on neuronal activity. In addition, BDNF caused a significant increase in glycine- and GABA-evoked current densities that partly explains the increase in synaptic transmission. Presynaptic mechanisms were also involved in BDNF effects since triethylammonium(propyl)-4-(2-(4-dibutylamino-phenyl)vinyl)pyridinium (FM1-43) destaining with high K(+) was augmented in neurons incubated with the neurotrophin. The effects of BDNF were mediated by receptor tyrosine kinase B (TrkB) and mitogen-activated protein kinase kinase (MEK) activation since culturing neurons with either (9S,10R,12R)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'- kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid methyl ester (K252a) or 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98059) blocked the augmentation in synaptic activity induced by the neurotrophin.

Anti-homeostatic synaptic plasticity of glycine receptor function after chronic strychnine in developing cultured mouse spinal neurons.

In this study, we describe a novel form of anti-homeostatic plasticity produced after culturing spinal neurons with strychnine, but not bicuculline or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Strychnine caused a large increase in network excitability, detected as spontaneous synaptic currents and calcium transients. The calcium transients were associated with action potential firing and activation of gamma-aminobutyric acid (GABA(A)) and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors as they were blocked by tetrodotoxin (TTX), bicuculline, and CNQX. After chronic blockade of glycine receptors (GlyRs), the frequency of synaptic transmission showed a significant enhancement demonstrating the phenomenon of anti-homeostatic plasticity. Spontaneous inhibitory glycinergic currents in treated cells showed a fourfold increase in frequency (from 0.55 to 2.4 Hz) and a 184% increase in average peak amplitude compared with control. Furthermore, the augmentation in excitability accelerated the decay time constant of miniature inhibitory post-synaptic currents. Strychnine caused an increase in GlyR current density, without changes in the apparent affinity. These findings support the idea of a post-synaptic action that partly explains the increase in synaptic transmission. This phenomenon of synaptic plasticity was blocked by TTX, an antibody against brain-derived neurotrophic factor (BDNF) and K252a suggesting the involvement of the neuronal activity-dependent BDNF-TrkB signaling pathway. These results show that the properties of GlyRs are regulated by the degree of neuronal activity in the developing network.

Therapeutic decision in anterior scleritis: our experience at a tertiary care eye center.

PURPOSE: To determine the characteristics of patients with anterior scleritis at a tertiary care eye center and determine which factors in these patients were associated with the need for systemic immunosuppressive therapy. PATIENTS AND METHODS: Retrospective study. Fifty patients with anterior scleritis presenting to the Cornea Service at Wills Eye Hospital from August 1996 to August 2001 were reviewed and divided into two groups. The control group included 35 patients who responded to oral non-steroid anti-inflammatory drugs (NSAIDs) and/or systemic steroids. The study group consisted of 15 patients who required additional treatment with systemic immunosuppressive agents. The influence of demographics, type of scleritis, bilateral disease, and ocular complications were analyzed. The presence of a systemic autoimmune disease, the best corrected visual acuity (BCVA), medications, and complications related to treatment were recorded. The statistical significance was evaluated by Student's t test for independence samples and Fisher's exact test; P<0.05 was considered significant. RESULTS: After a mean follow-up of 19.3 +/- 24.9 months, an associated systemic autoimmune disease was present in 17 patients (34%). Scleritis was the first manifestation of the systemic disease in 10 out of 17 patients (59%). Patients who required immunosuppressive treatment had more frequent ocular complications (80% versus 26%, P=0.0004) than did patients who responded to oral NSAIDs and/or steroids. CONCLUSIONS: Scleritis may be the sign of presentation of a systemic autoimmune disease. The presence of ocular complications, in particular corneal involvement, may be associated with the need for systemic immunosuppressive therapy.

Queratitis inflamatoria por acanthamoeba spp. análisis estructural por microscopía electrónica de transmisión / Infectious keratitis by acanthamoeba spp. amoeba structure. analysis using transmission electronics microscope

Se presenta el caso de una un paciente de 36 años, cuyo motivo de consulta fue visión borrosa y dolor ocular severo de 5 semanas de evolución, en ojo izquierdo. La paciente tenia como hábito el uso de lentes de contacto blandas con higiene inadecuada. Fue sometida a exámenes bacteriológicos y virológicos con resultados negativos. Para el presente estudio se extrajo material obtenido a través de una biopsia de córnea. Se realizó observación microscópica en fresco y con colorantes permanentes. Por medio de cultivos xénicos, se obtuvo trofozoitos y quistes de Acanthamoeba spp: Las colonias fueron observadas con microscopía óptica en fresco y con contraste de fases. Por MET (microscopía electrónica de transmisión) se analizó la ultraestructura de la ameba.

Botriomicosis seudotumoral / Pseudotumoral botryomycosis

La botriomicosis es una enfermedad purulenta crónica poco frecuente, de difícil diagnóstico clínico, que puede afectar a la piel y a otros órganos, caracterizada por una reaccción atípica del huésped frente a una infección bacteriana común, habitualmente por S. aureus. La histología muestra la formación de gránulos eosinofílicos que recuerdan a los observados en la actinomicosis. Describimos un caso de esta rara entidad en un paciente varón de 87 años que desarrolló una lesión nodular sobre el pliegue nasogeniano izquierdo con apariencia clínica similar a un queratoacantoma, en el que se llegó al diagnóstico tras realizar una exéresis y una biopsia de la lesión (AU)

IP(3) receptor function and localization in myotubes: an unexplored Ca(2+) signaling pathway in skeletal muscle.

We present evidence for an unexplored inositol 1,4,5-trisphosphate-mediated Ca(2+) signaling pathway in skeletal muscle. RT-PCR methods confirm expression of all three known isotypes of the inositol trisphosphate receptor in cultured rodent muscle. Confocal microscopy of cultured mouse muscle, doubly labeled for inositol receptor type 1 and proteins of known distribution, reveals that the receptors are localized to the I band of the sarcoplasmic reticulum, and this staining is continuous with staining of the nuclear envelope region. These results suggest that the receptors are positioned to mediate a slowly propagating Ca(2+) wave that follows the fast Ca(2+) transient upon K(+) depolarization. This slow wave, imaged using fluo-3, resulted in an increase in nucleoplasmic Ca(2+) lasting tens of seconds, but not contraction; the slow wave was blocked by both the inositol trisphosphate receptor inhibitor 2-aminoethoxydiphenyl borate and the phospholipase C inhibitor U-73122. To test the hypothesis that these slow Ca(2+) signals are involved in signal cascades leading to regulation of gene expression, we assayed for early effects of K(+) depolarization on mitogen-activated protein kinases, specifically extracellular-signal related kinases 1 and 2 and the transcription factor cAMP response element-binding protein (CREB). Within 30-60 seconds following depolarization, phosphorylation of both the kinases and CREB was evident and could be inhibited by 2-aminoethoxydiphenyl borate. These results suggest a signaling system mediated by Ca(2+) and inositol trisphosphate that could regulate gene expression in muscle cells.

Prognostic value of the glycoprotein TAG-72 in patients with gastric cancer.

The specificity of the tumor markers used to date in patients with gastric cancer has not been satisfactory. For this reason we decided to evaluate the utility of TAG-72 in this disease. Between 1993 and 1998 we determined the levels of TAG-72 in 638 subjects (148 healthy volunteers, 33 patients with chronic renal failure (CRF), 149 patients with benign diseases of the liver, 95 patients with benign gastrointestinal diseases and 213 patients with gastric cancer). TAG-72 was measured using an IRMA method. Statistical analysis of the data was performed with the BMDP package. We established a cutoff for TAG-72 of 3 U/mL, corresponding to the 92.6th percentile of the healthy controls. We observed that neither CRF nor benign liver diseases affected TAG-72 levels, while certain benign gastrointestinal diseases did cause alterations of the marker. Using Cox multivariate analysis we discovered that the preoperative TAG-72 level was an independent prognostic variable associated with both disease-free and overall survival. We conclude that, although TAG-72 is not useful for the diagnosis of gastric cancer, it is a suitable tool for disease monitoring and prognostic assessment.

Ciliary body tumor and cataract: local resection combined with phacoemulsification.

A 57-year-old man developed a ciliary body mass, clinically diagnosed as malignant melanoma of the ciliary body, that produced a cataract in the right eye. Treatment was cataract surgery with sclerouvectomy performed simultaneously. Pathohistologic examination revealed an acquired adenoma of the nonpigmented ciliary epithelium. The clinicopathologic features and treatment of this tumor are discussed.

Calcium transients in 1B5 myotubes lacking ryanodine receptors are related to inositol trisphosphate receptors.

Potassium depolarization of skeletal myotubes evokes slow calcium waves that are unrelated to contraction and involve the cell nucleus (Jaimovich, E., Reyes, R., Liberona, J. L., and Powell, J. A. (2000) Am. J. Physiol. 278, C998-C1010). Studies were done in both the 1B5 (Ry53-/-) murine "dyspedic" myoblast cell line, which does not express any ryanodine receptor isoforms (Moore, R. A., Nguyen, H., Galceran, J., Pessah, I. N., and Allen, P. D. (1998) J. Cell Biol. 140, 843-851), and C(2)C(12) cells, a myoblast cell line that expresses all three isoforms. Although 1B5 cells lack ryanodine binding, they bind tritiated inositol (1,4,5)-trisphosphate. Both type 1 and type 3 inositol trisphosphate receptors were immuno-located in the nuclei of both cell types and were visualized by Western blot analysis. After stimulation with 47 mm K(+), inositol trisphosphate mass raised transiently in both cell types. Both fast calcium increase and slow propagated calcium signals were seen in C(2)C(12) myotubes. However, 1B5 myotubes (as well as ryanodine-treated C(2)C(12) myotubes) displayed only a long-lasting, non-propagating calcium increase, particularly evident in the nuclei. Calcium signals in 1B5 myotubes were almost completely blocked by inhibitors of the inositol trisphosphate pathway: U73122, 2-aminoethoxydiphenyl borate, or xestospongin C. Results support the hypothesis that inositol trisphosphate mediates slow calcium signals in muscle cell ryanodine receptors, having a role in their time course and propagation.

Second intention healing after Mohs micrographic surgery.

BACKGROUND: Although second intention healing has been used for many years in Mohs micrographic surgery, it has lost popularity. This is due to the long process and functional alteration of important anatomical units if certain wound healing factors are not carefully taken into consideration. OBJECTIVE: To review indications, contraindications, complications, advantages, disadvantages, as well as basic concepts of the wound healing process, wound care and antibiotic prophylaxis in the management of open wounds after Mohs surgery. METHODS: A short but comprehensive review of studies published in the literature dealing with second intention healing as a reconstruction alternative option in certain situations. CONCLUSIONS: Second intention healing is a simple and cost-effective method for reconstruction after Mohs micrographic surgery in well-defined cases. It allows adequate tumoral control with good to excellent functional and cosmetic results.