Spatial Measurement and Inhibition of Calpain Activity in Traumatic Brain Injury with an Activity-Based Nanotheranostic Platform.

Traumatic brain injury (TBI) is a major public health concern that can result in long-term neurological impairments. Calpain is a calcium-dependent cysteine protease that is activated within minutes after TBI, and sustained calpain activation is known to contribute to neurodegeneration and blood-brain barrier dysregulation. Based on its role in disease progression, calpain inhibition has been identified as a promising therapeutic target. Efforts to develop therapeutics for calpain inhibition would benefit from the ability to measure calpain activity with spatial precision within the injured tissue. In this work, we designed an activity-based nanotheranostic (ABNT) that can both sense and inhibit calpain activity in TBI. To sense calpain activity, we incorporated a peptide substrate of calpain flanked by a fluorophore/quencher pair. To inhibit calpain activity, we incorporated calpastatin peptide, an endogenous inhibitor of calpain. Both sensor and inhibitor peptides were scaffolded onto a polymeric nanoscaffold to create our ABNT. We show that in the presence of recombinant calpain, our ABNT construct is able to sense and inhibit calpain activity. In a mouse model of TBI, systemically administered ABNT can access perilesional brain tissue through passive accumulation and inhibit calpain activity in the cortex and hippocampus. In an analysis of cellular calpain activity, we observe the ABNT-mediated inhibition of calpain activity in neurons, endothelial cells, and microglia of the cortex. In a comparison of neuronal calpain activity by brain structure, we observe greater ABNT-mediated inhibition of calpain activity in cortical neurons compared to that in hippocampal neurons. Furthermore, we found that apoptosis was dependent on both calpain inhibition and brain structure. We present a theranostic platform that can be used to understand the regional and cell-specific therapeutic inhibition of calpain activity to help inform drug design for TBI.

Sperm DNA Fragmentation in Male Infertility: Tests, Mechanisms, Meaning and Sperm Population to Be Tested.

Sperm DNA fragmentation (sDF) is a DNA damage able to predict natural conception. Thus, many laboratories added tests for the detection of sDF as an adjunct to routine semen analysis with specific indications. However, some points related to sDF are still open. The available tests are very different each from other, and a direct comparison, in terms of the prediction of reproductive outcomes, is mandatory. The proposed mechanisms responsible for sDF generation have not yielded treatments for men with high levels of sDF that have gained the general consent in clinical practice, thus requiring further research. Another relevant point is the biological meaning to attribute to sDF and, thus, what we can expect from tests detecting sDF for the diagnosis of male infertility. SDF can represent the "tip of iceberg" of a more extended and undetected sperm abnormality somehow impacting upon reproduction. Investigating the nature of such a sperm abnormality might provide novel insights into the link between sDF and reproduction. Finally, several studies reported an impact of native sDF on assisted reproduction technique outcomes. However, to fertilise the oocyte, selected spermatozoa are used where sDF, if present, associates with highly motile spermatozoa, which is the opposite situation to native semen, where most sDF associates with non-viable spermatozoa. Studies comparing the impact of sDF, as assessed in both native and selected spermatozoa, are needed.

Effects of cortisone in zebrafish (Danio rerio): Insights into gut microbiota interactions and molecular mechanisms underlying DNA damage and apoptosis.

Cortisone can enter aquatic ecosystems and pose a risk to organisms therein. However, few studies have explored the effects of cortisone on the gut microbiota of aquatic organisms. Here, we exposed zebrafish (Danio rerio) to cortisone at environmentally relevant concentrations (5.0, 50.0, or 500.0 ng L-1) for 60 days to explore its toxicological effects and their association with gut microbiota changes. The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling assay revealed that exposure to 50 ng L-1 cortisone significantly increased the intestinal cell apoptosis rate, 8-hydroxydeoxyguanosine contents, and caspase-3 and caspase-8 activities. Moreover, the transcriptome analysis results demonstrated a notable downregulation in the expression of most differentially expressed genes associated with apoptosis pathways, as well as changes in DNA replication, oxidative stress, and drug metabolism pathways; these results indicated the occurrence of cortisone-induced stress response in zebrafish. Molecular docking analysis revealed that cortisone can bind to caspase-3 through hydrogen bonds and hydrophobic interactions but that no such interactions occur between cortisone and caspase-8. Thus, cortisone may induce oxidative DNA damage and apoptosis by activating caspase-3. Finally, the 16S rRNA sequencing results demonstrated that cortisone significantly affected microbial community structures and functions in the intestinal ecosystem. These changes may indicate gut microbiota response to cortisone-induced intestinal damage and inflammation. In conclusion, the current results clarify the mechanisms underlying intestinal response to cortisone exposure and provide a basis for evaluating the health risks of cortisone in animals.

Suppressing Pro-Apoptotic Proteins by siRNA in Corneal Endothelial Cells Protects against Cell Death.

Corneal endothelial cells (CE) are critical for the cornea's transparency. For severe corneal damage, corneal tissue transplantation is the most promising option for restoring vision. However, CE apoptotic cell death occurs during the storage of donor corneas for transplantation. This study used small interfering (si)RNA-mediated silencing of pro-apoptotic proteins as a novel strategy to protect CE against apoptosis. Therefore, the pro-apoptotic proteins Bax and Bak were silenced in the human corneal endothelial cell line (HCEC-12) by transfection with Accell™siRNA without any adverse effects on cell viability. When apoptosis was induced, e.g., etoposide, the caspase-3 activity and Annexin V-FITC/PI assay indicated a significantly reduced apoptosis rate in Bax+Bak-siRNA transfected HCECs compared to control (w/o siRNA). TUNEL assay in HCECs exposed also significantly lower cell death in Bax+Bak-siRNA (7.5%) compared to control (w/o siRNA: 32.8%). In ex vivo donor corneas, a significant reduction of TUNEL-positive CEs in Bax+Bak-siRNA corneas (8.1%) was detectable compared to control-treated corneas (w/o siRNA: 27.9%). In this study, we demonstrated that suppressing pro-apoptotic siRNA leads to inhibiting CE apoptosis. Gene therapy with siRNA may open a new translational approach for corneal tissue treatment in the eye bank before transplantation, leading to graft protection and prolonged graft survival.

Analysis of Surgical Procedures on the Forearm and Hand and Their Relationship with Complex Regional Pain Syndrome: A Cross-sectional Study / Análise dos procedimentos cirúrgicos no antebraço e na mão e sua relação com a síndrome da dor regional complexa: Um estudo transversal

Abstract Objective Complex regional pain syndrome (CRPS) requires further understanding. Thus, the present study aimed to analyze if pre- and intraoperative factors may be related to the development of CRPS in the postoperative period. Methods We reviewed 1,183 medical records of patients undergoing forearm and hand surgeries from 2015 to 2021. The data of interest, that is, diagnosis, incisions, synthesis material, and anesthesia, were collected, tabulated, and statistically analyzed, with subsequent calculation of the odds ratios. Results Most patients were female, aged between 30 and 59 years, and sought the service electively (67% of the cases). The diagnoses included soft tissue trauma (43%), bone trauma (31.6%), and compressive syndromes (25.5%). During this period, 45 (3.8%) subjects developed CRPS. The statistical analysis showed that the chance of developing CRPS is twice as high in patients with compressive syndrome, especially carpal tunnel syndrome (CTS), which represented most surgeries performed in our service (24%). Two or more incisions occurred in 7.6% of the cases, which tripled the chance of developing postoperative CRPS. Gender, age, use pf synthetic material, type of anesthesia type did not statistically increase the risk of developing postoperative CRPS. Conclusion In short, the incidence of CRPS is low; however, it is critical to know and recognize the risk factors for prevention and active screening in the postoperative period.

Resumo Objetivo A síndrome da dor regional complexa (SDRC) precisa ser mais bem compreendida. Assim, este estudo objetiva analisar se fatores pré e intraoperatórios poderiam estar relacionados ao desenvolvimento de SDRC no pós-operatório. Métodos Foram revisados 1.183 prontuários de pacientes submetidos a cirurgias no antebraço e na mão entre 2015 e 2021. Os dados de interesse, como diagnóstico, incisões, material de síntese e anestesia realizada, foram coletados, tabulados e submetidos a testes estatísticos com posterior cálculo da razão de chances. Resultados A maioria dos pacientes era do gênero feminino, com idade entre 30 e 59 anos, que buscaram o serviço de forma eletiva (67% dos casos). Os diagnósticos agrupados de forma geral foram: traumas de partes moles (43%), traumas ósseos (31,6%) e síndromes compressivas (25,5%). Durante esse período, 45 pacientes (3,8%) evoluíram com SDRC. A análise estatística mostrou que a chance de desenvolver SDRC é duas vezes maior em pacientes com síndrome compressiva, especialmente a síndrome do túnel do carpo (STC), que representou a maioria dos cirurgias realizadas em nosso serviço (24%). Em 7,6% dos casos, foram realizadas duas ou mais incisões, o que triplicou a possibilidade de SDRC pósoperatória. Gênero, idade, uso de material de síntese, ou tipo de anestesia não aumentaram estatisticamente o risco de SDRC no pós-operatório. Conclusão Em suma, a incidência de SDRC é baixa, mas é importante conhecer e reconhecer os fatores de risco para a prevenção e a busca ativa no pós-operatório.

Apoptosis of hippocampus and cerebellum induced with brain ischemia reperfusion prevented by 3',4'-dihydroxyflavonol (DiOHF).

The present study aimed to determine the effect of 3',4'-dihydroxyflavonol (DiOHF) on apoptosis in the cerebellum and hippocampus in rats with ischemia-reperfusion. A total of 38 Wistar albino male rats were used. Experimental groups were designed as Group 1-Sham; Group 2-Ischemia-reperfusion (IR), in which animals were anesthetized and carotid arteries ligated for 30 minutes (ischemia) and reperfused 30 minutes; Group 3- IR + DiOHF (10 mg/kg); Group 4- Ischemia + DiOHF (10 mg/kg) + reperfusion; Group 5-DiOHF + IR. DiOHF was supplemented as 10 mg/kg by intraperitoneal injection 30 minutes before IR. Following application, the animals were sacrificed under general anesthetic by cervical dislocation, and the cerebellum and hippocampus tissues were analyzed for apoptosis. IR significantly increased hippocampus and cerebellum apoptosis activity, confirmed by Hematoxylin-Eosin, TUNEL labeling, and Caspase-8 activity. However, these values were significantly suppressed by the administration of DiOHF, especially when used before the ischemia and reperfusion. The results of the study show that increased apoptosis in the cerebellum and hippocampus tissue was inhibited by intraperitoneal DiOHF supplementation.

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.

Mechanism of isorhynchophylline in lipopolysaccharide-induced acute lung injury based on proteomic technology.

Isorhynchophylline (IRN), a tetracyclic indole alkaloid, has anti-inflammatory and antioxidant activities against cardiovascular diseases and central nervous system disorders. Acute lung injury (ALI) is a manifestation of inflammation concentrated in the lungs and has a high incidence rate and mortality The purpose of this study is to explain the mechanism of IRN in the treatment of acute lung injury and to provide a new scheme for clinical treatment. The experimental mice were divided into three groups: CTRL, LPS, LPS+IRN. The mouse model of ALI was established by inhaling LPS solution through nose. After continuous administration of IRN solution for 7 days, the mice in LPS+IRN group were killed and the lung tissue was collected for detection. Proteomic (Data are available via ProteomeXchange with identifier PXD050432) results showed that 5727 proteins were detected in mouse lung tissues, and 16 proteins were screened out. IRN could reverse the trend of these differential proteins. In addition, IRN can act on integrin αM to reduce neutrophil recruitment and thereby produce anti-inflammatory effects and may suppress neutrophil migration through the leukocyte transendothelial migration pathway. TUNEL and RT-PCR experiments revealed that LPS-induced ALI in mice increases the apoptosis of lung tissues, damage to alveolar epithelial cells and levels of inflammatory factors. Treatment with IRN can repair tissues, improve lung tissue pathology and reduce lung inflammation.

Sperm Chromatin Dispersion Test Detects Sperm DNA Fragmentation Mainly Associated with Unviable Spermatozoa and Underestimates the Values with Respect to TUNEL Assay.

Several clinical laboratories assess sperm DNA fragmentation (sDF) in addition to semen analysis in male infertility diagnosis. Among tests evaluating sDF, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) and SCD (Sperm Chromatin Dispersion) are widely used. Our lab developed a modified version of TUNEL (TUNEL/PI) able to distinguish two sperm populations (PI Brighter and PI Dimmer) differently associated with sperm viability and reproductive outcomes. The aim of this study was to compare sDF levels detected by SCD and TUNEL/PI in the semen samples from 71 male subjects attending our Andrology Laboratory. Our results demonstrate that SCD is less sensitive in determining sDF compared to TUNEL/PI. The statistically significant positive correlation found between sDF evaluated by SCD and PI Dimmer (consisting of all dead spermatozoa) suggests that SCD mainly detects sDF in unviable spermatozoa. We confirmed that most spermatozoa detected by SCD are unviable by performing SCD after incubation in hypo-osmotic medium to discriminate viable and unviable cells in 52 samples. Such results might explain the lower ability of this test in discriminating couples having successful ART outcomes demonstrated in published metanalyses. Overall, our results indicate that SCD is less sensitive in evaluating sDF for diagnostic purposes.

Use of Ozone Therapy and Thymoquinone in the Prevention of Formaldehyde Toxicity by Inhalation: An Experimental Study.

INTRODUCTION: The study determined the damage caused by formaldehyde (FA) exposure in blood and liver samples using biochemical markers. Histopathological analysis was performed using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method and measurement of CD68 cell density. To what extent the antioxidant molecules thymoquinone (TQ) and ozone (O3) reversed the damage caused by FA exposure was investigated, both when used alone and combined. METHODS: Fifty-six Sprague-Dawley male rats of eight to ten weeks of age were used in the experiment. The rats were divided into eight groups, with seven rats in each group: the untreated control group, the group treated with TQ (10 mg/kg/day), the group treated with O3 (150 µg/kg/day), the group treated with TQ+O3, the group exposed to FA (10 ppm 8 h/day), the group receiving FA+TQ, the group receiving FA+O3, and the group receiving FA+TQ+O3. Serum aspartate transaminase (AST), alanine transaminase (ALT), total antioxidant (TAS, U/mL), and total oxidant (TOS, nmol/mL) levels were analyzed. TAS and TOS levels, CD68 cell density, and apoptotic cells were determined in liver tissues. RESULTS: FA exposure caused an increase in serum AST and ALT levels of (p<0.05) experimental animals, a decrease in TAS levels in serum (p=0.03) and liver (p>0.05) and an increase in TOS levels (p>0.05), TUNEL positivity (p<0.001), and CD68 cell density (p=0.004). Administration of TQ and O3 as antioxidants significantly reversed biochemical and histopathological alterations in the serum and liver. CONCLUSION: TQ and ozone therapy suppressed oxidative stress caused by FA exposure and reversed the emerging histopathological deteriorations. Ozone therapy did not suppress the effects of TQ. Therefore, ozone therapy can be given as a supportive therapy along with the main therapeutic agents. We think TQ and ozone therapy may be useful to protect individuals exposed to FA.

TUNEL-n-DIFL Method for Detection and Estimation of Apoptosis Specifically in Neurons and Glial Cells in Mixed Culture and Animal Models of Central Nervous System Diseases and Injuries.

Detection of merely apoptosis does not reveal the type of central nervous system (CNS) cells that are dying in the CNS diseases and injuries. In situ detection and estimation of amount of apoptosis specifically in neurons or glial cells (astrocytes, oligodendrocytes, and microglia) can unveil valuable information for designing therapeutics for protection of the CNS cells and functional recovery. A method was first developed and reported from our laboratory for in situ detection and estimation of amount of apoptosis precisely in neurons and glial cells using in vitro and in vivo models of CNS diseases and injuries. This is a combination of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and double immunofluorescent labeling (DIFL) or simply TUNEL-n-DIFL method for in situ detection and estimation of amount of apoptosis in a specific CNS cell type. An anti-digoxigenin (DIG) IgG antibody conjugated with 7-amino-4-methylcoumarin-3-acetic acid (AMCA) for blue fluorescence, fluorescein isothiocyanate (FITC) for green fluorescence, or Texas Red (TR) for red fluorescence can be used for in situ detection of apoptotic cell DNA, which is earlier labeled with TUNEL using alkali-stable DIG-11-dUTP. A primary anti-NeuN (neurons), anti-GFAP (astrocytes), anti-MBP (oligodendrocytes), or anti-OX-42 (microglia) IgG antibody and a secondary IgG antibody conjugated with one of the above fluorophores (other than that of ani-DIG antibody) are used for in situ detection of apoptosis in a specific CNS cell type in the mixed culture and animal models of the CNS diseases and injuries.

Confocal Microscopy Technique in Teratogenicity Testing Using Zebrafish (Danio rerio) Embryos as Model.

Teratogenicity refers to the ability to cause adverse effects on the normal development of embryos resulting in retardation of growth as well as structural and functional abnormalities in the developing embryos. Zebrafish (Danio rerio) is one of the prime model organisms for teratogenicity testing, owing to the many advantages it offers, particularly its relatively large and initially transparent embryos, which allow real-time imaging of the various developmental stages. Confocal microscopy provides the best technique for imaging cellular dynamics within zebrafish embryos as it gives high-resolution imaging of thick tissues. This chapter focuses on major teratogenicity testing techniques using confocal microscopy. Terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) assay, immunohistochemistry assay, and reactive oxygen species (ROS) detection are important methods for studying the teratogenicity of drugs or compounds using 6 h post-fertilization Zebrafish embryos.

Evolución clínica en pacientes con síndrome del túnel carpiano tratados con ultrasonido o infiltración con corticoesteroides / Clinical Evolution in Patients with Carpal Tunnel Syndrome Treated with Ultrasound or Corticosteroid Infiltration

Introducción. El Síndrome del túnel carpiano es la neuropatía periférica compresiva más común de la extremidad superior, que se produce por la compresión del nervio mediano. Los casos leves y moderados pueden tratarse con métodos conservadores como ultrasonido terapéutico o infiltración con corticoesteroides. Objetivo. Describir la evolución clínica de pacientes con síndrome de túnel carpiano tratados con terapia por ultrasonido e infiltración de corticoesteroides. Metodología. Ensayo clínico abierto, en pacientes con síndrome del túnel carpiano leve y moderado, que consultaron del 1 de octubre 2021 al 30 de mayo 2022. Se formaron dos grupos; el que recibió tratamiento con ultrasonido con 12 casos y el grupo tratado con infiltración con corticoesteroides con seis casos. Ambos grupos fueron intervenidos en la consulta inicial, y luego, en las cuatro y ocho semanas posteriores al inicio del tratamiento. Resultados. Se muestran los resultados descriptivos relacionados con la intensidad de dolor, valorada con la Escala Visual Numérica, la infiltración obtuvo dos casos sin dolor y cuatro con dolor moderado, contrario a ultrasonido que se mantuvo con cuatro casos leves, tres moderados y cinco intensos. En los síntomas, la infiltración redujo el número de casos en cuatro de los síntomas estudiados, en cambio el ultrasonido únicamente en dos. En severidad, valorada con el cuestionario de Boston para túnel carpal, con infiltración se obtuvieron dos casos asintomáticos y ninguno con ultrasonido. Respecto a los signos clínicos, el signo de Tinel desapareció en cuatro casos en ambos grupos, mientras que signo de Phalen desapareció en cuatro casos en ultrasonido y dos en infiltración. Conclusión. En intensidad de dolor y grado de severidad, la infiltración generó casos asintomáticos y redujo mayor cantidad de síntomas que el ultrasonido. Ambos tratamientos disminuyeron la presencia de signos clínicos

Introduction. Carpal tunnel syndrome is the most common compressive peripheral neuropathy of the upper extremity, which is caused by compression of the median nerve. Mild and moderate cases can be treated with conservative methods such as therapeutic ultrasound or corticosteroid infiltration. Objective. To describe the clinical evolution of patients with carpal tunnel syndrome treated with ultrasound therapy and corticosteroid infiltration. Methodology. A prospective open clinical trial was conducted in patients with mild and moderate carpal tunnel syndrome who consulted from October 1, 2021 to May 30, 2022. Two groups were formed: the group that received ultrasound treatment with 12 cases and the group treated with corticosteroid infiltration with six cases. Both groups were treated at the initial consultation and then at four and eight weeks after the start of treatment. Results. The descriptive results related to the intensity of pain, evaluated with the Visual Numeric Scale, are shown. Infiltration obtained two cases without pain and four with moderate pain, contrary to ultrasound which was maintained with four mild, three moderate and five intense cases. In symptoms, infiltration reduced the number of cases in four of the symptoms studied, while ultrasound reduced the number of cases in only two. In severity, assessed with the Boston carpal tunnel questionnaire, with infiltration, there were two asymptomatic cases and none with ultrasound. Regarding clinical signs, Tinel's sign disappeared in four cases in both groups, while Phalen's sign disappeared in four cases in ultrasound and two in infiltration. Conclusion. Infiltration produced asymptomatic patients and reduced more symptoms than ultrasonography in terms of pain intensity and severity. Clinical symptoms were less common with both treatments.

[Analysis of the echographic parameters described in the study of carpal tunnel syndrome: A systematic review]. / Análisis de los parámetros ecográficos descritos en el estudio del síndrome del túnel del carpo. Una revisión sistemática.

Carpal tunnel syndrome (CTS) is the most common entrapment mononeuropathy; the diagnosis is established by electrodiagnostic tests with until 34% of false positives/negatives. We present the following systematic review which objective is to analyze the most recent literature related to the ultrasound parameters described to study CTS. We selected studies that evaluated ultrasound parameters in patients with clinical suspicion following the Cochrane manual's recommendations. We include systematic reviews, meta-analyses, case-control studies and diagnostic tests, evaluating retrospective studies and bibliographic reviews with proper methodological quality. Articles published between 2005 and 2019. We included eight articles (two systematic reviews/meta-analyses, two case-control studies, one diagnostic test study, two literature reviews, and one retrospective). The parameters analyzed were cross-sectional area, wrist-forearm index, entry-exit index, thinning range, palmar bowing of the flexor retinaculum, and vascularity/mobility. Current evidence allows us to affirm that ultrasound is useful in screening for CTS.

Sensory and Motor Median Nerve Neuropathy Due to a Deep Giant Hand Lipoma: A Case Report / Neuropatia sensorial e motora do nervo mediano por um lipoma gigante na mão: Relato de caso

Abstract Lipomas are the most common soft-tissue tumors in the human body, but their location in the hand is rare. Symptomatic hand lipomas, due to nerve compression, are even rarer. We present a case of median nerve neuropathy as a result of a giant palm lipoma, located on the thenar and hypothenar areas of the hand. The patient had typical symptoms of carpal tunnel syndrome, along with compromised thumb motion. Intraoperatively, the recurrent motor branch of the median nerve was sitting on the lipoma under a great tension. This particular location of the motor branch of the median nerve in relation to the lipoma makes this case unique. The tumor was excised protecting the neurovascular structures, and a few weeks later the patient regained full thumb motion, grip strength, and resolution of dysesthesia.

Resumo Os lipomas são os tumores de partes moles mais comuns no corpo humano, mas sua localização na mão é rara. Os lipomas de mão que causam sintomas por compressão do nervo são ainda mais raros. Apresentamos um caso de neuropatia do nervo mediano decorrente de um lipoma palmar gigante, localizado nas regiões tenar e hipotenar da mão. A paciente apresentava sintomas típicos de síndrome do túnel do carpo, além de comprometimento dos movimentos do polegar. Durante a cirurgia, o ramo motor recorrente do nervo mediano repousava sobre o lipoma sob grande tensão. Esta localização particular do ramo motor do nervo mediano em relação ao lipoma torna este caso único. O tumor foi extirpado, protegendo as estruturas neurovasculares e, poucas semanas depois, a paciente havia recuperado totalmente os movimentos do polegar e força de preensão, além de apresentar resolução da disestesia.

Usefulness of the Phalen Test and the Tinel Sign in the Prognosis and the Impact on Quality of Life of Patients with Carpal Tunnel Syndrome Undergoing Classical Open Carpal Tunnel Release / Utilidade do teste de Phalen e do sinal de Tinel no prognóstico e o impacto na qualidade de vida de pacientes com síndrome do túnel do carpo submetidos a liberação aberta clássica do túnel do carpo

Abstract Objective: To evaluate the usefulness of the Phalen test and the Tinel sign in the prognosis and the impact on quality of life in the clinical course of patients with carpal tunnel syndrome undergoing surgical treatment through the traditional open approach. Methods: The present is a cohort study on prognosis. We included 115 patients with high probability of receiving a clinical diagnosis of carpal tunnel syndrome with indication for surgical treatment. All patients underwent the Phalen test and Tinel sign and answered the Boston Carpal Tunnel Questionnaire before and after the surgical treatment. Results: The estimates for the probability of the time until remission of the Phalen test at 2, 4 and 16 weeks postoperatively were of 3.54% (95% confidence interval [95% CI]: 1.16%-8.17%), 0.88% (95%CI: 0.08%-4.38%) and 0.88% (95%CI: 0.08% to 4.38%) respectively, and, for the Tinel sign, they were of 12.39% (95%CI: 7.13%-19.18%), 4.42% (95%CI : 1.65%-9.36%) and 2.65% (95%CI : 0.70%-6.94%) respectively. There was a reduction in the postoperative score on the Boston Carpal Tunnel Questionnaire of 1.8 points for symptom severity (p < 0.001) and of 1.6 points for functional status (p < 0.001). Conclusion: Phalen test remission was earlier than that of the Tinel sign, but, when performed as of the second postoperative week, they were prognostic factors favorable to the clinical course, with improved quality of life.

Resumo Objetivo: Avaliar a utilidade do teste de Phalen e do sinal de Tinel no prognóstico e o impacto na qualidade de vida no curso clínico de pacientes com síndrome do túnel do carpo submetidos ao tratamento cirúrgico por via aberta clássica. Métodos: Trata-se de um estudo de coorte sobre prognóstico. Foram incluídos 115 pacientes com alta probabilidade de diagnóstico clínico de síndrome do túnel do carpo com indicação de tratamento cirúrgico. Todos os pacientes foram submetidos ao teste de Phalen e ao sinal de Tinel, e responderam ao questionário de Boston antes e depois do tratamento cirúrgico. Resultados: As estimativas de probabilidade do tempo até a remissão do teste de Phalen em 2, 4 e 16 semanas pós-operatórias foram de 3,54% (intervalo de confiança de 95% [IC95%]:1,16%-8,17%), 0,88% (IC95%: 0,08%-4,38%) e 0,88% (IC95%: 0,08%-4,38%), respectivamente, e, do sinal de Tinel, foram de 12,39% (IC95%: 7,13%-19,18%), 4,42% (IC95%: 1,65%-9,36%) e 2,65% (IC95%: 0,70%-6,94%), respectivamente. Na pontuação pós-operatória no Questionário de Boston, houve redução de 1,8 ponto para a gravidade dos sintomas (p < 0,001), e de 1,6 ponto para o estado funcional (p < 0,001). Conclusão: A remissão do teste de Phalen foi mais precoce do que a do sinal de Tinel, mas, realizados a partir da segunda semana de evolução pós-operatória, esses testes foram fatores prognósticos favoráveis ao curso clínico, com melhora da qualidade de vida.

Long-term Changes of Infrared Thermography in Successful Decompression of Carpal Tunnel Syndrome: A Case Report / Alterações a longo prazo da termografia infravermelha na descompressão bem-sucedida da Síndrome do Túnel do Carpo: Relato de caso

Introduction Infrared thermography (IRT) has demonstrated high diagnostic accuracy for carpal tunnel syndrome (CTS) diagnosis in previous studies. However, the recovery of the autonomic function after treatment for CTS is rarely addressed in the literature, especially on the long-term. Case Presentation A 59-year-old lady sought treatment for a long-term history of numbness, tingling, and hand and arm pain. CTS was diagnosed by clinical and electrophysiological means. After 6 months of conservative treatment, surgical treatment was offered. Preoperative IRT was performed by static and dynamic evaluations immediately and 5 minutes after the cold challenge test using the FLIR C2 camera with accuracy of 2°C or 2%. Fingers were consistently colder (mean of 3.76° C), which clearly represented an autonomic dysfunction in the patient's hand. The patient underwent mini-open carpal tunnel decompression and did great postoperatively. One year after surgery, the patient was fully recovered and completely asymptomatic. IRT imaging showed a remarkable improvement of fingers temperature (mean of 3.36°C). Conclusion Our long-term results confirmed that functional recovery occurred concomitantly to autonomic recovery, which was demonstrated by consistent improvement in fingers' temperature. IRT has a strong potential at the evaluation of patients with CTS for both diagnosis and follow-up.

Introdução A termografia infravermelha (IRT) demonstrou alta precisão diagnóstica para o diagnóstico da síndrome do túnel do carpo (STC) em estudos anteriores. No entanto, a recuperação da função autonômica após o tratamento para STC é raramente abordada na literatura, especialmente a longo prazo. Apresentação do caso Uma senhora de 59 anos procurou tratamento para um histórico de longo prazo de dormência, formigamento e dor nas mãos e braços. A STC foi diagnosticada por meios clínicos e eletrofisiológicos. Após 6 meses de tratamento conservador, o tratamento cirúrgico foi oferecido. A IRT pré-operatória foi realizada por avaliações estáticas e dinâmicas imediatamente e 5 minutos após o teste de provocação pelo frio usando a câmera FLIR C2 com precisão de 2 °C ou 2%. Os dedos estavam consistentemente mais frios (média de 3,76 °C), o que claramente representava uma disfunção autonômica na mão da paciente. A paciente foi submetida a uma mini descompressão aberta do túnel do carpo e teve um ótimo desempenho no pós-operatório. Um ano após a cirurgia, a paciente estava totalmente recuperada e completamente assintomática. A imagem IRT mostrou uma melhora notável na temperatura dos dedos (média de 3,36 °C). Conclusão Nossos resultados de longo prazo confirmaram que a recuperação funcional ocorreu concomitantemente à recuperação autonômica, o que foi demonstrado pela melhora consistente na temperatura dos dedos. A IRT tem um forte potencial na avaliação de pacientes com STC para diagnóstico e acompanhamento.

Neuroprotective Effects of Human Adipose-Derived Mesenchymal Stem Cells in Oxygen-Induced Retinopathy.

This study was designed to provide evidence of the neuroprotective of human adipose-derived mesenchymal stem cells (hADSCs) in oxygen-induced retinopathy (OIR). In vivo, hADSCs were intravitreally injected into OIR mice. Various assessments, including HE (histological evaluation), TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) staining, electroretinogram (ERG) analysis, and retinal flat-mount examination, were performed separately at postnatal days 15 (P15) and 17 (P17) to evaluate neurological damage and functional changes. Western blot analysis of ciliary neurotrophic factor (CNTF), glial cell line-derived neurotrophic factor (GDNF), and brain-derived neurotrophic factor (BDNF) was conducted at P17 to elucidate the neuroprotective mechanism. The P17 OIR group exhibited a significant increase in vascular endothelial cell nuclei and neovascularization that breached the ILM (inner limiting membrane) to the P17 control group. In addition, the retinal nonperfusion areas in the P17 OIR group and the number of apoptotic retinal cells in the P15 OIR group were significantly higher than in the corresponding hADSCs treatment group and control group. There was no significant thickness change in the inner nuclear layer (INL) but the outer nuclear layer (ONL) in the P17 OIR treatment group compared with the P17 OIR group. The cell density in the INL and ONL at P17 in the hADSCs treatment group was not significantly different from the OIR group. The amplitude of a-wave and b-wave in scotopic ERG analysis for the P17 OIR group was significantly lower than in the P17 hADSCs treatment group and the P17 control group. Furthermore, the latency of the a-wave and b-wave in the P17 OIR group was significantly longer than in the P17 hADSCs treatment group and the P17 control group. In addition, the expression levels of CNTF and BDNF in the P17 OIR group were statistically higher than those in the P17 control group, whereas the expression of GDNF was statistically lower in the P17 OIR group, compared with the P17 control group. The expression of CNTF and GDNF in the P17 hADSCs treatment group was statistically higher than in the P17 OIR group. However, the expression of BDNF in the P17 hADSCs treatment group was statistically lower than in the P17 OIR group. This study provides evidence for the neuroprotective effects of hADSCs in OIR.

Bisphenol A exposure decreases sperm production and male fertility through inhibition PCBP2 expression.

Growing evidence suggests that the exposure of bisphenol A (BPA), an endocrine disruptor that commonly present in the environment, can impair reproduction. However, conflicting results have been reported, and the underlying mechanism has not been fully understood. In this study, 3-week-old male mice were oral exposed to 50 mg/kg/d BPA or equivalent corn oil for 28 days. Their testis and epididymis were then collected for morphology examination by HE stains. The number of sperm was counted, and the morphology was analyzed by PNA (peptide nucleic acid) and pap staining. Fertilization capacity and successful rate were analyzed after mating with wide-type females. Spermatid DNA damage and apoptosis were evaluated by DFI, γH2AX stain, and TUNEL assay. RNA sequencing analysis was conducted to identify differentially expressed genes in testicular tissue of mice exposed to BPA. RNA interference was used to verify the regulatory mechanism of BPA exposure on gene expression in GC-2 cells. Our data showed that the total number of sperm was decreased and the morphology was impaired in BPA-exposed mice. In addition, the serum testosterone level and fertilization efficiency were also reduced. Mechanism studies showed that BPA could suppress the expression of PCBP2, a key regulatory gene in spermatid development, by activating the EZH2/H3K27me3. In conclusion, we found that BPA exposure can impair spermatid development via affecting key gene expression that is at least partially due to epigenetic modification.

Multi-Effects of Acute Salinity Stress on Osmoregulation, Physiological Metabolism, Antioxidant Capacity, Immunity, and Apoptosis in Macrobrachium rosenbergii.

Salinity stress can trigger a series of physiological changes. However, the mechanism underlying the response to acute salinity stress in Macrobrachium rosenbergii remains poorly understood. In this study, osmoregulation, physiological metabolism, antioxidant capacity, and apoptosis were examined over 96 h of acute salinity stress. Hemolymph osmolality increased with increasing salinity. After 48 h of salinity exposure, the glucose, triglycerides, total protein, and total cholesterol contents in two salinity stress groups (13 and 26‱ salinity) were significantly lower than those in the 0‱ salinity group. The highest levels of these parameters were detected at 6 h; however, superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) were the lowest at 96 h in the 13‱ salinity group. The activity of immunity-related enzyme alkaline phosphatase (AKP) showed a decreasing trend with increasing salinity and remained at a low level in the 26‱ salinity group throughout the experiment. No significant differences were observed in aspartate aminotransferase (AST), alanine aminotransferase (ALT), or lysozyme (LZM) among the three treatments at 96 h. After 96 h of salinity treatments, the gill filament diameter significantly decreased, and a more pronounced terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive signal was detected in the 13‱ and 26‱ groups compared to that in the 0‱ group. Expression levels of apoptosis-related genes, including Cysteine-aspartic acid protease 3 (Caspase 3), Cysteine-aspartic acid protease 8 (Caspase 8), Cytochrome c (Cyt-c), tumor suppressor gene (P53), Nuclear factor kappa-B (NF-κB), and B cell lymphoma 2 ovarian killer (Bok) were significantly higher in the 26‱ salinity group than in the other groups at 24 h, but lower than those in the 0‱ salinity group at 96 h. Cyt-c and P53 levels exhibited a significantly positive relationship with MDA, AST, and LZM activity during salinity stress. In the 13‱ salinity group, Bok expression was significantly correlated with SOD, T-AOC, AKP, acid phosphatase, and LZM activity, whereas in the 26‱ group, the AST content was positively correlated with Caspase 8, Cyt-c, and P53 expression. A significant negative relationship was observed between Caspase 3 expression and catalase (CAT) activity. These findings provide insight into the mechanisms underlying the response to acute salinity stress and will contribute to improving M. rosenbergii aquaculture and management practices.