Metal-organic Framework Immobilized Enzyme and Its Application in Screening of Enzyme Inhibitors of Traditional Chinese Medicine：A Review / 中国实验方剂学杂志

Enzymes are widely used in chemical and pharmaceutical industries because of their advantages of high efficiency and specificity. However， the shortcomings of the free enzymes， such as poor stability and difficulty in recycling， limit their application. Therefore， the immobilization and application of enzymes have become one of the research hotspots. The selection of the immobilization carriers is a critical step in the process of enzyme immobilization. Metal-organic frameworks（MOFs）， a kind of porous materials， are formed by the coordination of metal ions or metal clusters with organic ligands. As an emerging immobilization carrier， its advantages such as high porosity， strong stability， and surface modifiability make it ideal for immobilized enzyme carriers. By immobilizing the free enzyme on MOFs， the above mentioned deficiencies of the free enzymes can be effectively solved， which greatly broaden the applicable condition. Ligand fishing is a method to find receptor-specific ligands from complex components， which has the advantages of high efficiency， simple sample pretreatment and high specificity. The MOF-enzyme complex formed by enzyme immobilization can act as a "fishing rod" for ligand fishing， which can screen out the targets from the complex system of components. The complex chemical composition and various active ingredients of traditional Chinese medicine（TCM） make the ligand fishing technology to play a big role in the screening of enzyme inhibitors from TCM. And the screened enzyme inhibitors are expected to be further developed into the lead compounds with good efficacy and low adverse effects， so the immobilized enzymes of MOFs have a wide application in the screening of active ingredients from TCM. Based on this， this paper summarized the methods of immobilized enzymes of MOFs in recent years， analyzed the characteristics， advantages and disadvantages of each method， and summarized the laws of preparation conditions and mechanisms. Meanwhile， the application and future development of immobilized enzymes of MOFs in the field of enzyme inhibitor screening from TCM were also summarized and prospected， with a view to providing a reference for the development of natural ingredients and the modernization of TCM.

Application of the ICU Mobility Scale in patients submitted to cardiac surgery / Aplicação da ICU Mobility Scale em pacientes submetidos à cirurgia cardíaca

Abstract Introduction Early mobilization is an alternative used in the Intensive Care Unit (ICU) to reduce the effects of immobility. The Intensive Care Unit Mobility Score (IMS) is applied to assess mobility status. Objective To determine the functional level of ICU patients submitted to cardiac surgery using the IMS scale. Methods This is an analytical observational study carried out with adult ICU patients submitted to cardiac surgery. Data on the use of vasoactive drugs, functional level through IMS, duration of mechanical ventilation and length of ICU stay were collected from the physiotherapy routine form and recorded on a specific instrument developed for the study. The data obtained from the IMS scale score on the postoperative days in the ICU were used to classify the patients' mobility during the hospitalization period. Descriptive statistics were used to present the data. Results A total of 69 patients were evaluated, 43% of whom were men, and all had an SMI of 0 on the first postoperative day. Classification on the scale increased over the course of hospitalization (IMS between 7 and 10), despite the use of vasoactive drugs in 54.6% of the individuals. Conclusion Patients submitted to cardiac surgery admitted to the ICU had moderate-to-high mobility levels throughout their stay and at discharge from the ICU.

Resumo Introdução A mobilização precoce é uma alternativa utilizada em Unidades de Terapia Intensiva (UTIs) na tentativa de reduzir os efeitos decorrentes do imobilis-mo. A escala de mobilidade em UTIs ou Intensive Care Unit Mobility Score (IMS) é aplicada para avaliar o nível de mobilidade. Objetivo Verificar por meio da escala IMS o nível de funcionalidade de pacientes submetidos à cirurgia cardíaca internados em uma UTI. Métodos Trata-se de um estudo observacional analítico realizado em UTI com pacientes adultos submetidos à cirurgia car-díaca. Foram coletados da ficha de rotina da fisioterapia informações quanto ao uso de drogas vasoativas, nível funcional por meio da IMS, tempo de ventilação mecânica e de internação na UTI, e registrados em instrumento específico desenvolvido para o estudo. Os dados obtidos do escore da escala IMS nos dias de pós-operatório na UTI foram utilizados para classificar a mobilidade dos pacientes durante o período de internação. Utilizou-se estatística descritiva para a apresentação dos dados. Resultados Foram avaliados 69 pacientes, 43% eram do sexo masculino e todos apresentavam IMS 0 no primeiro dia de pós-operatório. A classificação na escala aumentou com o decorrer do período de internação (IMS entre 7 e 10), apesar do uso de drogas vasoativas em 54,6%. Conclusão Os pacientes submetidos à cirurgia cardíaca internados na UTI apresentaram níveis de mobilidade de moderado a alto ao longo da internação e na alta da UTI.

Cacería para autoconsumo durante la pandemia de COVID-19: el caso de las comunidades Matsigenkas del Parque Nacional del Manu

En este trabajo, se hipotetiza el periodo de aislamiento, la inmovilización y la migración inversa que sucedidos durante la pandemia del COVID-19 (años 2020 y 2021) incrementaron la capacidad de caza de las comunidades Matsigenkas asentadas en el Parque Nacional del Manu (PN Manu). Para esto, evaluamos los cambios en los patrones del volumen de extracción de la caza a través de un monitoreo participativo utilizando el modelo de monitoreo de la intensidad de cacería. Se evidenció que hubo un incremento de la capacidad de caza de las comunidades nativas. Los volúmenes más altos de extracción y el mayor incremento del número de cazadores, esfuerzo y área de cacería se dieron en el periodo 2020 - 2021, cuando las medidas de aislamiento e inmovilización fueron establecidas. Sin embargo, hubo una disminución de los indicadores para la temporada 2021 - 2022, año en que se flexibilizaron las medidas de aislamiento y se dio la reactivación económica, permitiendo que las personas retornaran a sus actividades económicas y educativas fuera del PN Manu. Se debe considerar que los efectos de solo un año de pandemia sobre los medios de subsistencia de las comunidades pudieron afectar la abundancia de la fauna silvestre, donde el mono maquisapa y mono choro (especies vulnerables) tuvieron altos incrementos de extracción. Por otro lado, nuestros resultados indicarían que el buen estado de conservación del PN Manu permitió la provisión de alimentos a las comunidades durante la pandemia. Se recomienda la planificación de programas que mitiguen los efectos negativos de pandemias o eventos semejantes, además se señala la importancia de medir el grado de resiliencia de las principales especies de caza, ya que el incremento de caza también podría tener efectos sobre la seguridad alimentaria de estas comunidades.

In this work, we hypothesize that the period of isolation, immobilization, and reverse migration that occurred during the COVID-19 pandemic (years 2020 and 2021) increased the hunting capacity of the Matsigenka communities settled in Manu National Park (PN Manu). To evaluate this, we assessed changes in hunting volume patterns through participatory monitoring using the hunting intensity monitoring model. It was evident that there was an increase in the hunting capacity of Matsigenka communities. The highest extraction volumes and the greatest increase in the number of hunters, hunting effort, and hunting area occurred in the 2020-2021 period, the year in which isolation and immobilization measures were established. However, there was a decrease in the indicators for the 2021-2022 period, the year in which isolation measures were relaxed and economic reactivation occurred, allowing people to return to their economic and educational activities outside PN Manu. It should be considered that the effects of only one year of the pandemic on the livelihoods of the communities could have affected the abundance of wildlife, with vulnerable species such as maquisapa monkeys and choro monkeys experiencing high extraction rates. On the other hand, our results indicate that the good conservation status of PN Manu provided food provisions for the communities during the pandemic. Planning programs to mitigate the negative effects of pandemics or similar events is recommended, and the importance of measuring the resilience of key hunting species is emphasized, as increased hunting could also have effects on the food security of these communities.

Restrição do movimento da coluna: uma análise do conhecimento dos profissionais de enfermagem / Restricted movement of the spine: an analysis of the knowledge of nursing professionals / Restricción del movimiento de la columna vertebral: un análisis del conocimiento de los profesionales de enfermería

RESUMO: Objetivo: Analisar o conhecimento de profissionais de enfermagem sobre restrição do movimento da coluna em vítimas de trauma e descrever a tomada de decisão. Método: Estudo exploratório, descritivo, quali-quantitativo. Realizou-se entrevista semiestruturada com 27 profissionais de enfermagem de serviços de emergência hospitalar e pré-hospitalar em São Leopoldo, Brasil, em março de 2022. Os dados foram analisados por estatística descritiva e análise de conteúdo. Resultados: 48% dos profissionais tinham conhecimento quanto às recomendações atuais; a maioria atuantes no pré-hospitalar. A cervicalgia/lombalgia foi a indicação predominante. Na categoria "a tomada de decisão e a transição do cuidado", identificou-se falta de padronização nas condutas do intra-hospitalar, enfatizando a importância da sequência no atendimento e confiança entre profissionais. Considerações finais: Identificou-se pouca apropriação das equipes do intra-hospitalar sobre as atuais evidências, falta de padronização nas condutas e dificuldades na transição do cuidado entre os serviços.

ABSTRACT Objective: To analyze nursing professionals' knowledge about restricting spinal movement in trauma victims and describe their decision-making. Method: An exploratory, descriptive, qualitative-quantitative study. A semi-structured interview was conducted with 27 nursing professionals from hospital and pre-hospital emergency services in São Leopoldo, Brazil, in March 2022. The data was analyzed using descriptive statistics and content analysis. Results: 48% of the professionals knew the current recommendations; most worked in pre-hospital care. Cervicalgia/lumbago was the predominant indication. In the category "decision making and the transition of care", a lack of standardization in in-hospital conduct was identified, emphasizing the importance of sequence in care and trust between professionals. Final considerations: It was found that in-hospital teams had little knowledge of the current evidence, a lack of standardization in procedures, and difficulties in the transition of care between services.

RESUMEN Objetivo: Analizar los conocimientos de los profesionales de enfermería sobre la restricción del movimiento de la columna vertebral en víctimas de traumatismos y describir su toma de decisiones. Método: Un estudio exploratorio, descriptivo, cualitativo-cuantitativo. Se realizó una entrevista semiestructurada a 27 profesionales de enfermería de los servicios de urgencias hospitalarios y prehospitalarios de São Leopoldo, Brasil, en marzo de 2022. Los datos se analizaron mediante estadística descriptiva y análisis de contenido. Resultados: El 48% de los profesionales conocía las recomendaciones actuales; la mayoría de ellos trabajaba en atención prehospitalaria. La cervicalgia/lumbago fue la indicación predominante. En la categoría "toma de decisiones y transición de los cuidados", se detectó una falta de normalización en la conducta intrahospitalaria, lo que subraya la importancia de la secuencia en los cuidados y la confianza entre los profesionales. Consideraciones finales: Se identificó que los equipos intrahospitalarios tenían escaso conocimiento de las evidencias actuales, que había una falta de normalización en los procedimientos y dificultades en la transición de la atención entre los servicios.

Evaluation of the comparative effects of infrared and red laser photobiomodulation therapy on skeletal muscle atrophy in an immobilization model in rats

Introduction: Skeletal muscle atrophy leads to a reduction in muscle strength, functionality, and the quality of life of individuals. Objective: To explore the effects of two different wavelengths (red and infrared) of laser PBMT on muscle atrophy and its active ingredients on skeletal muscle atrophy using an in vivo model of muscle atrophy. Methods: Thirty-two Wistar rats were randomly divided into four experimental groups: control (CG) animals were not immobilized and did not receive any type of treatment; immobilized animals with no treatment (ImC); immobilized animals submitted to red laser with wavelength of 660 nm (ImR) and near-infrared laser with wavelength of 808 nm (ImIR) treatments. The treatments were applied daily, at 2 points in the right gastrocnemius muscle (cranial and caudal), through the punctual contact technique, for 9 sessions, with the first application immediately after removing the cast. Results: The histological results demonstrated that in both treated groups (red and infrared wavelengths) a reduction of the inflammatory infiltrate and less connective tissue thickening when compared to the ImC. However, only infrared light was observed regenerating muscle fibers and an increase in the number of oxidative fibers (type I). Conclusion: These results suggest that red and infrared wavelength laser PBMT were able to promote changes in the morphology of the gastrocnemius muscle submitted to atrophy in an experimental immobilization model, reducing the inflammatory infiltrate and the formation of intramuscular connective tissue. However, infrared laser PBMT promoted more evident positive effects by increasing regenerating muscle fibers and the number of oxidative fibers.

Effect of acute or subchronic stress on T cell response in peripheral blood: Regulatory role of vitamin D

The immune response, orchestrated by helper (Th1, Th2, and Th17) and regulatory (Treg) T cells, is modulated by stress and Vitamin D (Vit-D). Although the immunomodulatory functions of both are known, their specific roles on Th cells have not been fully clarified, yet. On this background, we aimed to investigate the effect of acute or subchronic stress on the distribution of peripheral T lymphocytes, as well as the immunomodulatory role of Vit-D. Young adult male, Swiss-albino mice (30–40g) were allocated to the control, acute stress (AS), subchronic stress (ChS), control+Vit-D, AS+Vit-D, and ChS+Vit-D groups (n=11/group). The combined cold (2-h at 4°C)-immobilization (2-h in a restrainer) stress protocol was employed as one day in AS groups and five consecutive days in ChS groups. Vit-D (2?g/kg ip) was applied every other day, until the end of the protocol. Serum cortisol, Vit-D and cytokine levels (IL-4, IFN-?, and IL-17A) were measured, and lymphocytes from blood samples were subtyped by flow-cytometry. Stress exposure caused differential Th and Treg responses, acute stress shifting the response to Th1, and subchronic stress shifting the response to Th2. Th17 and Treg cells were lower in subchronic stress exposed mice. These changes became comparable to control values in Vit-D treated groups. The T cell response, crucial for immune system function, differs on the basis of stress exposure as such the Vit-D treatment. The tolerogenic profile created by Vit-D should be considered for management of stress-related diseases. Our results may help to provide a better understanding of disease pathogenesis.

Advances in 3D printing-based body membrane immobilization technology in radiotherapy / 中华放射医学与防护杂志

The widespread application of 3D printing and computer technologies in the medical field has opened up opportunities for digital and automated fabrication of body immobilization tools for radiotherapy, thus making it possible to get rid of the original complex manual fabrication process. As the most widely used technique for body immobilization, body membrane immobilization has always attracted much attention. This review outlines the development of the body membrane immobilization technology in different radiotherapy stages. Moreover, the advances in the application of 3D-printed body membranes were introduced, as a development direction of body immobilization technology, in the field of radiotherapy. This technology can be utilized as a reference for clinical application and research.

Dosimetric effects of prone immobilization devices combined with a belly-board in intensity-modulated radiotherapy for gynecologic cancers / 中华放射医学与防护杂志

Objective:To investigate the dosimetric effects of prone immobilization devices combined with a belly board (PIDBBs) in the intensity-modulated radiotherapy (IMRT) for gynecologic cancers.Methods:A total of 20 patients with cervical or endometrial cancer treated with radiotherapy in the Third Affiliated Hospital of Sun Yat-sen University from August 2020 to June 2021 were retrospectively analyzed. Two sets of body contours were outlined for each patient. One set of body contours did not contain the immobilization devices, and the other contour set included the immobilization devices. For each patient, doses were calculated for the two sets of contours using the same 7-field IMRT plan and were recorded as Plan without and Plan with. The dosimetric difference caused by the immobilization devices was assessed by comparing the parameter values in the dose-volume histograms (DVHs) and by plan subtraction. The Gafchromic EBT3 film and anthropomorphic phantom were used to verify the calculated doses. Results:The target coverage and average dose of Plan with were lower than those of Plan without. Specifically, the V50 Gy, V49 Gy, and Dmean of planning target volume (PTV) decreased by 19.75%, 7.99%, and 2.54% ( t = 8.96, 10.49, 22.09, P < 0.01), respectively. The V40 Gy, V30 Gy, V20 Gy, V15 Gy, and Dmean of skins increased by 51.79%, 51.05%, 45.72%, 33.63% and 10.80% ( t = -2.54, -5.63, -15.57, -24.06, -13.88, P < 0.01), respectively. Doses to other organs at risk (OARs) showed no significant differences. As indicated by the EBT3 measurements, the doses to skins of the abdomen and pelvis on the anthropomorphic phantom increased by approximately 37.24% ( t = 10.86, P<0.01). Conclusions:Although PIDBBs can effectively reduce the low dose to the small intestine, the radiation attenuation caused by them can reduce the PTV coverage of radiotherapy plans and increase the doses to abdominal and pelvic skins sharply, especially for patients requiring irradiation of the groin and perineum.

Immobilization of mutated xylanase from Neocallimastix patriciarum in E. coli and application for kraft pulp biobleaching / Imobilização de xilanase mutada de Neocallimastix patriciarum em E. coli e aplicação para polpa kraft

Abstract As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.

Resumo Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.

Development of enzyme immobilization systems for CO2 bioconversion: advances and challenges / 生物工程学报

Enzyme-catalyzed CO2 reduction to value-added commodities is important for alleviating the global environmental issues and energy crises due to high selectivity and mild conditions. Owing to high energy density, formic acid or methanol produced from CO2 using formate dehydrogenase (FDH) or multi-enzyme cascades are promising target chemicals for CO2 utilization. However, the low activity, poor stability and low reusability of key enzymes involved in such process hampered its large-scale application. Enzyme immobilization provides an effective solution to these problems and significant progress have been made in immobilization carriers. Moreover, integration of enzyme immobilization with other catalysis techniques have been explored extensively. This review summarized the recent advances in the immobilization of enzymes using membranes, inorganic materials, metal-organic frameworks, covalent organic frameworks and other carriers, and illustrated the characteristics and advantages of different immobilization materials and immobilization methods. The synergistic effects and applications of immobilized enzymes and electrocatalytic or photocatalytic coupling reaction systems for CO2 reduction were further summarized. Finally, the current challenges of enzyme immobilization technology and coupling reaction systems were pointed out and their development prospects were presented.

Histological analysis of the effects of electroacupuncture stimulation on the femur fracture line in a rat model of hindlimb immobilization / 全日本鍼灸学会雑誌

[Introduction] This study compared the effects of joint immobilization and concurrent electroacupuncture stimulation on fracture line appearance, which is indicative of differences in cortical bone structure, in the femurs of young adult rats. [Materials and Methods] Forty-two 7-week-old Wistar rats were divided into three groups: a hindlimb immobilized group (IM), a hindlimb immobilized group with concurrent electroacupuncture stimulation (IMEA), and an untreated control group (CO). IM and IMEA were immobilized for two weeks with a jacket-type immobilization device that immobilized the hip joint in extension and restricted hip joint adduction and abduction. Acupuncture needles were inserted into the femoral region during the immobilization period in the IMEA group, and continuous alternating current was used for stimulation. A low-frequency stimulator was used to generate electrical current (250 μsec, 50 Hz, 0.24 mA, 500 Ω load resistance), and stimulation was performed daily for two weeks at 10 min/day. The extracted femur was fractured from the anterior part of the diaphysis or metaphyseal end by three-point-bending under constant conditions with 10 mm between the fulcrum and a crosshead speed of 10 mm/min. [Results] IM showed significantly lower Stiffness, Deformation, and Strength values than CO. Bone morphometry parameters also showed lower values for IM. A cement line was observed between lamellar and non-lamellar bone in CO and IMEA, but was indistinct in IM. In CO and IMEA, minute bone fragments were formed in the deep diaphysis region sandwiched between the inner and outer circumferential lamellae, but no such fragments were found in IM. IMEA had less resorption than IM, and lamellar bone with a structure similar to CO was maintained. Moreover, cracks observed in CO and IMEA ran longitudinally in the vicinity of the cement line and rest line, but no such cracks were found in IM. [Discussion and Conclusion] These findings suggest that electroacupuncture mitigates bone weakening in immobilized rat femurs, and this affects the appearance of the fracture line.

Immobilizing engineered Escherichia coli cells into zeolitic imidazolate framework 8 for efficient biosynthesis of Ala-Gln / 生物工程学报

The α-amino acid ester acyltransferase (SAET) from Sphingobacterium siyangensis is one of the enzymes with the highest catalytic ability for the biosynthesis of l-alanyl-l-glutamine (Ala-Gln) with unprotected l-alanine methylester and l-glutamine. To improve the catalytic performance of SAET, a one-step method was used to rapidly prepare the immobilized cells (SAET@ZIF-8) in the aqueous system. The engineered Escherichia coli (E. coli) expressing SAET was encapsulated into the imidazole framework structure of metal organic zeolite (ZIF-8). Subsequently, the obtained SAET@ZIF-8 was characterized, and the catalytic activity, reusability and storage stability were also investigated. Results showed that the morphology of the prepared SAET@ZIF-8 nanoparticles was basically the same as that of the standard ZIF-8 materials reported in literature, and the introduction of cells did not significantly change the morphology of ZIF-8. After repeated use for 7 times, SAET@ZIF-8 could still retain 67% of the initial catalytic activity. Maintained at room temperature for 4 days, 50% of the original catalytic activity of SAET@ZIF-8 could be retained, indicating that SAET@ZIF-8 has good stability for reuse and storage. When used in the biosynthesis of Ala-Gln, the final concentration of Ala-Gln reached 62.83 mmol/L (13.65 g/L) after 30 min, the yield reached 0.455 g/(L·min), and the conversion rate relative to glutamine was 62.83%. All these results suggested that the preparation of SAET@ZIF-8 is an efficient strategy for the biosynthesis of Ala-Gln.

Immobilization of mutated xylanase from Neocallimastix patriciarum in E coli and application for kraft pulp biobleaching / Imobilização de xilanase mutada de Neocallimastix patriciarum em E coli e aplicação para polpa kraft

As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.

Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.

Immobilization of mutated xylanase from Neocallimastix patriciarum in E. coli and application for kraft pulp biobleaching

Abstract As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.

Resumo Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.

Estresse juvenil de curta frequência e intensidade não afeta a substância branca cerebral de ratos / Juvenile stress of short frequency and intensity does not affect rats´ brain white matter

O objetivo deste estudo foi avaliar os efeitos duradouros de dois tipos de estresse sobre o corpo caloso (CC). Foram estudados 42 ratos Wistar machos divididos aleatoriamente em três grupos: Grupo Controle (GC), Estresse Físico (EF, imobilização) e Estresse Psicológico (EP, exposição ao predador). Os procedimentos de estresse ocorreram durante três dias consecutivos na idade juvenil (P25-P27) e foram analisados na idade adulta (P74). Os cérebros foram coletados, processados com a técnica de Klüver-Barrera, e secções foram analisadas por meio de morfometria. Os resultados demonstraram que não houve alterações em aspectos gerais como peso dos animais, e histológicos como espessura do CC e quantidade dos núcleos gliais nesta região. O estudo sugere que os efeitos duradouros de ambos os modelos de estresse juvenil de curta frequência (3 dias) e intensidade (90 minutos/EF e 20 minutos/EP) não foram nem prejudiciais e nem protetores, o que pode ser considerado uma adaptação positiva.

The current study evaluates the lasting effects of two types of stress on the corpus callosum (CC). Forty-two male Wistar rats were randomly divided into three groups: Control Group (CG), Physical Stress (FS, immobilization) and Psychological Stress (PS, exposure to predators). Stress procedures occurred for three consecutive days at the juvenile stage (P25-P27) and analyzed at the adult age (P74); brains were retrieved and processed by Klüver-Barrera technique and sections were analyzed by morphometry. Results showed that there were no changes in the general aspects such as animal weight, and in the histological aspects such as CC thickness and quantity of the region´s glia nuclei. Current research suggests that the lasting effects of both models of juvenile stress of short frequency (3 days) and intensity (90 minutes/FS and 20 minutes/PS) were neither detrimental nor protective, featuring a positive adaptation.

Técnica com retalho muscular duplo e fasciocutâneo na correção de úlceras isquiáticas por pressão / Technique with double muscle and fasciocutaneous flap in the correction of sciatic pressure ulcers

Úlceras por pressão são lesões ocasionadas na pele e tecidos subjacentes devido à força de pressão local, geralmente em pontos de proeminências ósseas. Cita-se aqui o caso de uma paciente acamada devido à lesão medular por mielomeningocele que evoluiu com úlcera em região isquiática à direita, tratada com técnica de retalhos muscular e fasciocutâneo de face posterior da coxa. Por ser lesão rotineiramente encontrada nesses pacientes portadores de limitações funcionais, é fundamental a realização de tratamentos adequados que visem a melhora clínica do paciente e minimizar índice de recidivas. Além disso, é de suma importância a implementação de novas técnicas cirúrgicas, haja vista a enorme variedade de lesões por pressão

Pressure ulcers are injuries to the skin and underlying tissues due to local pressure force, usually at points of bony prominence. We mention here the case of a bedridden patient due to a spinal cord injury caused by myelomeningocele that evolved with an ulcer in the right sciatic region, treated with the muscle flap and fasciocutaneous flap technique of the posterior thigh. As it is a lesion routinely found in these patients with functional limitations, it is essential to carry out appropriate treatments aimed at improving the patients clinical condition and minimizing the rate of recurrences. In addition, the implementation of new surgical techniques is of utmost importance, given the enormous variety of pressure injuries

A comparative study of free laccase and laccase immobilized in copper alginate

Enzyme immobilization has gained considerable attention due to the incredible properties exhibited by the enzymes in immobilized condition. Therefore, in the present study, a comparative analysis of free and immobilized laccase is reported using various substrates. The ideal substrate for immobilization was found to be copper alginate with an immobilization yield of 91.078%. The optimum pH was 4 and 5, respectively, for free and immobilized enzymes while the optimum temperature was found to be 50 and 60?C, respectively. The kinetic parameters Vmax and Km were found from the Line weaver-Burk plot and were 48.076 U/mL and 0.480 mM for free enzyme while 55.55 U/mL and 1.277 mM for the immobilized enzyme, respectively. The catalytic efficiency Kcat was found as 100.01s?1 for free enzyme and 43.5s?1 for its immobilized counterpart. Out of the various metal ions used, Co2+ was found to enhance the activity of an immobilized enzymes. The storage stability of the enzyme was studied and was found that only 32.44% of initial activity was retained by free enzyme whereas, 70.21% of activity was retained by immobilized enzyme upon storage for four weeks at 4?C. The thermal stability studies shows that the immobilized enzyme retained 32.60% of its initial activity and the free enzyme retained 1.14% of initial activity on exposure to 60?C for 3 h. Finally, the reusability of immobilized laccase beads was evaluated by decolorization of methyl orange for five repeated cycles and a percentage decolorization of 32.04% could be retained at the fifth cycle. This study therefore, suggests copper alginate-immobilized beads to be an effective option for various applications

A preparation strategy for protein-oriented immobilized silica magnetic beads with Spy chemistry for ligand fishing / 药物分析学报

Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.

Application of optical surface monitoring system in intra-fraction motion monitoring in frameless cranial stereotactic radiotherapy / 中华放射医学与防护杂志

Objective:To establish a novel clinical application process of the optical surface monitoring system (OSMS) in the cranial frameless stereotactic radiosurgery (SRS) and stereotactic radiotherapy (SRT), and to assess the accuracy and effectiveness of the OSMS in the intra-fraction motion monitoring of both cranial phantoms and cranial SRT patients fixed using the Q-Fix encompass immobilization system.Methods:The deviations of OSMS in the real-time motion monitoring were assessed by determining the deviations between the displacement of the cranial SRS phantoms detected by the OSMS and the predefined displacement of the Varian Edge six degrees of freedom (6DoF) couch. The ability of the OSMS to conduct real-time monitoring of the head movement was also analyzed when one camera was blocked by the rotary gantry of the accelerator and when the couch was at non-zero angles. Moreover, ten patients who received 50 fractions of cranial frameless SRT were enrolled in this study. All the patients were fixed using the Q-Fix Encompass system, and their intra-fraction motion was monitored using the OSMS. The intra-fraction errors of OSMS real-time monitoring throughout the treatment were obtained from the OSMS logs. The patients received cone-beam computed tomography (CBCT) after the beam delivery, and the six-dimensional errors were obtained as intra-fraction motion errors of the CBCT.Results:For the cranial phantoms, there was a close correlation between the OSMS monitoring deviations and the predefined displacement in six dimensions. The OSMS-detected 3D vector deviations in the translational and rotational directions were (0.28±0.10) mm and (0.15±0.09)°, respectively when the angel both the gantry and couch was 0° and were (0.35±0.13) mm and(0.17±0.09)°, respectively, when one camera was blocked. The OSMS monitoring deviations with the couch at a non-zero degree were greater than those at zero degree. The maximum deviations occurred when the couch was at 270° and were (0.69±0.19) mm and (0.32±0.12)°, respectively, in the translational and rotational directions. For the cranial SRT patients fixed using the Q-Fix Encompass system, the OSMS and CBCT showed comparable intra-fractional motion deviations, which were (0.40±0.26) and (0.29±0.10) mm, respectively in the translational direction and were (0.33±0.20)°and (0.26±0.08)° in the rotational direction.Conclusions:The OSMS is an effective tool for optically guided radiotherapy, which allows for intra-fraction real-time motion monitoring with sub-millimeter accuracy. Therefore, to ensure the accurate preformation of cranial SRS/SRT, it is necessary to conduct the intra-fractional position monitoring using OSMS.

Molecular engineering and immobilization of lysine decarboxylase for synthesis of 1, 5-diaminopentane: a review / 生物工程学报

1, 5-diaminopentane, also known as cadaverine, is an important raw material for the production of biopolyamide. It can be polymerized with dicarboxylic acid to produce biopolyamide PA5X whose performances are comparable to that of the petroleum-based polyamide materials. Notably, biopolyamide uses renewable resources such as starch, cellulose and vegetable oil as substrate. The production process does not cause pollution to the environment, which is in line with the green and sustainable development strategy. The biosynthesis of 1, 5-diaminopentane mainly includes two methods: the de novo microbial synthesis and the whole cell catalysis. Lysine decarboxylase as the key enzyme for 1, 5-diaminopentane production, mainly includes an inducible lysine decarboxylase CadA and a constituent lysine decarboxylase LdcC. Lysine decarboxylase is a folded type Ⅰ pyridoxal-5' phosphate (PLP) dependent enzyme, which displays low activity and unstable structure, and is susceptible to deactivation by environmental factors in practical applications. Therefore, improving the catalytic activity and stability of lysine decarboxylase has become a research focus in this field, and molecular engineering and immobilization are the mainly approaches. Here, the mechanism, molecular engineering and immobilization strategies of lysine decarboxylase were reviewed, and the further strategies for improving its activity and stability were also prospected, with the aim to achieve efficient production of 1, 5-diaminopentane.