Effects of rumen microorganisms on the decomposition of recycled straw residue / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Recently, returning straw to the fields has been proved as a direct and effective method to tackle soil nutrient loss and agricultural pollution. Meanwhile, the slow decomposition of straw may harm the growth of the next crop. This study aimed to determine the effects of rumen microorganisms (RMs) on straw decomposition, bacterial microbial community structure, soil properties, and soil enzyme activity. The results showed that RMs significantly enhanced the degradation rate of straw in the soil, reaching 39.52%, which was 41.37% higher than that of the control on the 30th day after straw return. After 30 d, straw degradation showed a significant slower trend in both the control and the experimental groups. According to the soil physicochemical parameters, the application of rumen fluid expedited soil matter transformation and nutrient buildup, and increased the urease, sucrase, and cellulase activity by 10%‒20%. The qualitative analysis of straw showed that the hydroxyl functional group structure of cellulose in straw was greatly damaged after the application of rumen fluid. The analysis of soil microbial community structure revealed that the addition of rumen fluid led to the proliferation of Actinobacteria with strong cellulose degradation ability, which was the main reason for the accelerated straw decomposition. Our study highlights that returning rice straw to the fields with rumen fluid inoculation can be used as an effective measure to enhance the biological value of recycled rice straw, proposing a viable solution to the problem of sluggish straw decomposition.

Enzyme production mechanism of anaerobic fungus Orpinomyces sp. YF3 in yak rumen induced by different carbon source / 生物工程学报

In order to investigate the enzyme production mechanism of yak rumen-derived anaerobic fungus Orpinomyces sp. YF3 under the induction of different carbon sources, anaerobic culture tubes were used for in vitro fermentation. 8 g/L of glucose (Glu), filter paper (Flp) and avicel (Avi) were respectively added to 10 mL of basic culture medium as the sole carbon source. The activity of fiber-degrading enzyme and the concentration of volatile fatty acid in the fermentation liquid were detected, and the enzyme producing mechanism of Orpinomyces sp. YF3 was explored by transcriptomics. It was found that, in glucose-induced fermentation solution, the activities of carboxymethyl cellulase, microcrystalline cellulase, filter paper enzyme, xylanase and the proportion of acetate were significantly increased (P < 0.05), the proportion of propionate, butyrate, isobutyrate were significantly decreased (P < 0.05). The results of transcriptome analysis showed that there were 5 949 differentially expressed genes (DEGs) between the Glu group and the Flp group, 10 970 DEGs between the Glu group and the Avi group, and 6 057 DEGs between the Flp group and the Avi group. It was found that the DEGs associated with fiber degrading enzymes were significantly up-regulated in the Glu group. Gene ontology (GO) function enrichment analysis identified that DEGs were mainly associated with the xylan catabolic process, hemicellulose metabolic process, β-glucan metabolic process, cellulase activity, endo-1,4-β-xylanase activity, cell wall polysaccharide metabolic process, carbohydrate catabolic process, glucan catabolic process and carbohydrate metabolic process. Moreover, the differentially expressed pathways associated with fiber degrading enzymes enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were mainly starch and sucrose metabolic pathways and other glycan degradation pathways. In conclusion, Orpinomyces sp. YF3 with glucose as carbon source substrate significantly increased the activity of cellulose degrading enzyme and the proportion of acetate, decreased the proportion of propionate, butyrate and isobutyrate. Furthermore, the degradation ability and energy utilization efficiency of fungus in the presence of glucose were improved by means of regulating the expression of cellulose degrading enzyme gene and participating in starch and sucrose metabolism pathway, and other glycan degradation pathways, which provides a theoretical basis for the application of Orpinomyces sp. YF3 in practical production and facilitates the application of Orpinomyces sp. YF3 in the future.

Analysis of the biocompatibility of a biocelulose and a poly L- lactic acid membrane

The use of selective barriers as resorbable membranes has become a routine clinical procedure for guided bone regeneration. Therefore, the production of membranes with a low inflammatory potential during their resorption process has become the goal of a considerable number of researches. Aim: The purpose of the present study was to evaluate the biocompatibility of poly (L- lactic acid) (PLLA) and biocelulose membranes (BC) inserted in the subcutaneous tissue on the dorsum of rats. Methods: Fifteen animals underwent surgical procedures for the insertion of 4 types of membranes: COL (Collagen membrane) ­ Control Group; BC (Biocellulose membrane); BCAg (Biocellulose membrane impregnated with Silver); PLLA (Poly (L-lactic acid) membrane). All membrane types were inserted into each animal. Animals were euthanized after 3, 7, and 15 days of the surgical procedure. Descriptive histological analyses were carried out to investigate host tissue reaction to membrane presence by assessing the anti-inflammatory process composition associated with the membrane resorption and the presence of foreign-body reaction or encapsulation. Results: The BC membranes showed a higher degree of inflammation and poor pattern of integration with the surrounding tissues than the PLLA and COL membranes. Conclusion: The PLLA and COL membranes present better biocompatibility than the BC membranes

Amplificación directa de ADN de Bordetella pertussis purificado de hisopados nasofaríngeos por un método de bajo costo, rápido (60-segundos) y libre de equipos / Direct amplification of Bordetella pertussis DNA purified from nasopharyngeal swabs by a low-cost, fast (60-second), and equipment-free method

RESUMEN Objetivo. Desarrollar y evaluar un método de bajo costo basado en celulosa para la purificación rápida y amplificación directa de ADN de Bordetella pertussis de hisopados nasofaríngeos. Materiales y métodos. Se prepararon discos de celulosa y se evaluaron diferentes parámetros (buffers de lisis/lavado, número de discos y elución de ADN). El método se acopló a una amplificación directa por PCR en tiempo real (qPCR) y se estimó el rendimiento utilizando hisopados nasofaríngeos que fueron positivos (n=100) y negativos (n=50) para ADN B. pertussis por qPCR, comparado con el método basado en columnas de sílice. Se calculó el grado de concordancia, sensibilidad, especificidad, valor predictivo positivo (VPP) y valor predictivo negativo (VPN). Se evaluó la factibilidad del método rápido para ser acoplado a un ensayo colorimétrico de amplificación isotérmica mediada por lazo (LAMP). Resultados. El método rápido con un disco de celulosa y buffer de lisis y lavado conteniendo PVP-40 y Tween 20, respectivamente, mostró una mayor capacidad para purificar ADN amplificable de B. pertussis. El método tuvo una sensibilidad de 89,0% (IC95%, 80,2%-94,9%) y una especificidad de 98,5% (IC95%, 92,1%-100,0%), con un buen grado de concordancia (Kappa=0,867; IC95% 0,788 - 0,946), respecto al método referencial. Los VPP y VPN fueron 98,6% (IC95%, 92,7,2%-100,0%) y 88,2% (IC95%, 78,7%-94,4%), respectivamente. Se evidenció una amplificación exitosa por LAMP, y se obtuvieron resultados comparables con el método por columnas de sílice. Conclusión. El método desarrollado es simple, de bajo costo y libre de equipos para la obtención rápida (60 segundos) de ADN en el punto de atención, y puede ser implementado en diversas técnicas moleculares orientados al diagnóstico oportuno y al estudio epidemiológico de tos ferina.

ABSTRACT Objective. To develop and evaluate a low-cost cellulose-based method for rapid purification and direct amplification of Bordetella pertussis DNA from nasopharyngeal swabs. Materials and methods. We prepared cellulose discs and evaluated different parameters (lysis/wash buffers, number of discs and DNA elution). The method was coupled to a direct real-time PCR (qPCR) amplification and the performance was estimated using nasopharyngeal swabs that were positive (n=100) and negative (n=50) for B. pertussis DNA by qPCR, compared to the silica column-based method. We calculated sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) and the degree of agreement. The feasibility of the rapid method to be coupled to a loop-mediated isothermal amplification colorimetric assay (LAMP) was evaluated. Results. The rapid method, with a cellulose disk and lysis and wash buffer containing PVP-40 and Tween 20, respectively, showed a greater capacity to purify amplifiable DNA from B. pertussis. The method had a sensitivity of 89.0% (95%CI: 80.2%-94.9%) and a specificity of 98.5% (95%CI: 92.1%-100.0%), with a good degree of agreement (Kappa=0.867; 95%CI: 0.788 - 0.946), compared to the reference method. The PPV and NPV were 98.6% (95%CI: 92.7.2%-100.0%) and 88.2% (95%CI: 78.7%-94.4%), respectively. Successful amplification by LAMP was evident, and comparable results were obtained with the silica column method. Conclusion. The developed method is simple, low-cost and equipment-free for rapid (60 seconds) DNA collection at the point of care, and can be implemented in various molecular techniques aimed at the timely diagnosis and epidemiological study of pertussis.

Immobilisation of solvatochromic dyes on transparent cellulose films; an improved methodology for the study of homeopathic potencies

Highly diluted and succussed solutions (homeopathic potencies) have been shown to interact with a wide range of solvatochromic dyes based on changes in their UV-visible spectra. Studies so far have involved free dyes in solution, but there is a pressing need to find ways to investigate the potency-dye interaction using isolated dye molecules in order to ask more searching physico-chemical questions regarding the fundamental nature of potencies. Aims and Methods:The aims of the present study have been tolook for ways to covalently immobilizesolvatochromic dyes onto transparent cellulose films and hence be in a position to investigate dye-potency interactions without the complication of dye-dye interactions, including dye aggregation, which can occur with free dyes in solution. Results: to date a total of nine different dyes have been immobilised on cellulose films using epoxide activation of hydroxyl groups on the cellulose surface. Using this methodology studies have begun looking at the time course of potency action on one of these immobilised dyes, Brooker's merocyanine. Results show that the interaction ofArsenicum10M with this dye consists of three phases ­an initial growth phase, a sustained plateau of interaction and a final decline phase lasting several days.Conclusions: amethodology has been developed that successfully immobilises solvatochromic dyes onto transparent cellulose film. These films can then be used in a spectrophotometer to study at a much more detailed level how potencies interact with dyes compared with using free dyes in solution. Results indicate that the information gained in this way provides new insights regarding the fundamental nature of potencies. Specifically, studies using immobilised Brooker's merocyanine with Arsenicum10M reveal that the lifetime of the potency is much longer than expected and that its action consists of three distinct phases, suggesting a resonant interaction with the dye.

Changes in chemical composition of cover crops residue during decomposition / Mudanças na composição química dos resíduos culturais de plantas de cobertura durante a decomposição

Crop residues decomposition are controlled by chemical tissue components. This study evaluated changes on plant tissue components, separated by the Van Soest partitioning method, during cover crop decomposition. The Van Soest soluble fraction was the first to be released from the crop residues, followed by cellulose and hemicellulose. Lignin was the crop residue component that suffered the least degradation, and for certain crop residue types, lignin degradation was not detected. The degradation of the main components of crop residues (soluble fraction, cellulose, hemicellulose and lignin) is determined by the chemical and structural composition of each fraction.

A decomposição de resíduos culturais é controlada pela composição química do tecido vegetal. O objetivo deste estudo foi avaliar as alterações que ocorrem nos componentes do tecido vegetal, separados pelo fracionamento de Van Soest, durante a decomposição de plantas de cobertura. A fração solúvel foi a primeira a ser liberada dos resíduos culturais, seguida pela celulose e hemicelulose. A lignina foi o componente dos resíduos culturais de menor degradação, sendo que em alguns resíduos culturais não foi possível detectar a degradação deste componente. A degradação dos principais componentes dos resíduos culturais (fração solúvel, celulose, hemicelulose e lignina) é determinada pela composição química e estrutural de cada uma destas frações.

Application of nanocellulose in flexible sensors / 生物医学工程学杂志

The shortage of medical resources promotes medical treatment reform, and smart healthcare is a promising strategy to solve this problem. With the development of Internet, real-time health status is expected to be monitored at home by using flexible healthcare systems, which puts forward new demands on flexible substrates for sensors. Currently, the flexible substrates are mainly traditional petroleum-based polymers, which are not renewable. As a natural polymer, cellulose, owing to its wide range of sources, convenient processing, biodegradability and so on, is an ideal alternative. In this review, the application progress of nanocellulose in flexible sensors is summarized. The structure and the modification methods of cellulose and nanocellulose are introduced at first, and then the application of nanocellulose flexible sensors in real-time medical monitoring is summarized. Finally, the advantages and future challenges of nanocellulose in the field of flexible sensors are discussed.

Development of an improved virus plaque assay based on avicel / 生物工程学报

Avicel is made of a mixture of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), and used for virus plaque assay. The avicel in common use is produced by FMC Biopolymer. Due to the relatively fixed proportion of MCC and CMC, avicel in common use is not suitable for plaque determination experiment of all types of viruses. In this study, we evaluated the effect of avicel made of different proportions of MCC and CMC on virus plaque assay, and developed an improved avicel virus plaque assay featured with simple and convenient operation, good practicability and high stability. To generate avicel overlays with different proportions of MCC and CMC, twelve different 2×avicel solutions were prepared. Their overall viscosity and bottom viscosity were measured to evaluate the ease of operation. The results showed that most of the 2×avicel solutions (except the 4.8% MCC+1.4% CMC and 4.8% MCC+1.0% CMC group) were easy to absorb and prepare nutrient overlap than 2×CMC solution. In order to find the best scheme to detect the titer of porcine epidemic diarrhea virus (PEDV), these avicel overlay solutions with different proportion of MCC and CMC were used as a replacement in the standard plaque assay. By comparing the size, clarity, stability and titer accuracy of virus plaque, we identified that 0.6% MCC and 0.7% CMC was the most preferable composition of avicel overlay for PEDV plaque assay. In conclusion, we developed an improved virus plaque assay based on avicel, which may facilitate the research of virus etiology, antiviral drugs and vaccines.

Regulating the structure of bacterial cellulose by altering the expression of bcsD using CRISPR/dCas9 / 生物工程学报

Gluconacetobacter xylinus is a primary strain producing bacterial cellulose (BC). In G. xylinus, BcsD is a subunit of cellulose synthase and is participated in the assembly process of BC. A series of G. xylinus with different expression levels of the bcsD gene were obtained by using the CRISPR/dCas9 technique. Analysis of the structural characteristics of BC showed that the crystallinity and porosity of BC changed with the expression of bcsD. The porosity varied from 59.95%-84.05%, and the crystallinity varied from 74.26%-93.75%, while the yield of BC did not decrease significantly upon changing the expression levels of bcsD. The results showed that the porosity of bacterial cellulose significantly increased, while the crystallinity was positively correlated with the expression of bcsD, when the expression level of bcsD was below 55.34%. By altering the expression level of the bcsD gene, obtaining BC with different structures but stable yield through a one-step fermentation of G. xylinus was achieved.

Enhancement of dissolution rate of racecadotril by liquisolid compact technology

Abstract The current investigation was used to improve the rate of dissolution of an anti-diarrheal drug i.e., racecadotril (RT) at low pH conditions (i.e., in the stomach) by reducing the water secretion and electrolyte in to the intestine by liquisolid tablets. Different formulations (liquisolid) were prepared using Avicel PH 102 as a carrier. Aerosil 200 as a coating material and sodium starch glycolate used as a disintegrant. Polyethylene glycol 200 was used as a non-volatile vehicle to dissolve the drug. FTIR, DSC, XRD and dissolution studies were conducted to characterise liquisolid tablets. Characterisation studies indicated that no interactions between carrier and drug. Solid state characterization had shown a reduction in crystallinity that further supports increment in solubility and dissolution. The optimised formulation showed a significant increase in dissolution i.e., 99.54±0.62% in 30 min compared to directly compressible tablets (38.47±0.26%). The % dissolution efficiency of racecadotril liquisolid tablets 76.86% compared to marketed tablet (27.56%) and conventional direct compression tablet (17.11%). Significant reduction in mean dissolution time of racecadotril from liquisolid tablets (6.84 min) compared to direct compression tablet (44.57 min), indicating faster release of drug and faster onset of action. Formulation of liquisolid tablets could enhance solubility, dissolution and bioavailability of racecadotril

Metagenomics approach to identify lignocellulose-degrading enzymes in the gut microbiota of the Chinese bamboo rat cecum

BACKGROUND: Lignocellulose is considered a renewable organic material, but the industrial production of biofuel from lignocellulose is challenging because of the lack of highly active hydrolytic enzymes. The guts of herbivores contain many symbiotic microorganisms that have evolved to hydrolyze plant lignocellulose. Chinese bamboo rats mainly consume high-fiber foods, indicating that some members of the intestinal tract microbiota digest lignocellulose, providing these rats with the energy required for growth. RESULTS: Here, we used metagenomics to analyze the diversity and functions of the gut microbiota in Chinese bamboo rats. We identified abundant populations of lignocellulose-degrading bacteria, whose main functions involved carbohydrate, amino acid, and nucleic acid metabolism. We also found 587 carbohydrate-active enzyme genes belonging to different families, including 7 carbohydrate esterase families and 21 glycoside hydrolase families. The glycoside hydrolase 3, glycoside hydrolase 1, glycoside hydrolase 43, carbohydrate esterase 4, carbohydrate esterase 1, and carbohydrate esterase 3 families demonstrated outstanding performance. CONCLUSIONS: The microbes and enzymes identified in our study expand the existing arsenal of proficient degraders and enzymes for lignocellulosic biofuel production. This study also describes a powerful approach for targeting gut microbes and enzymes in numerous industries.

Evaluation of self-prepared absorbable hemostatic cellulose fibrils / 浙江大学学报·医学版

To evaluate the effectiveness and safety of self-prepared absorbable hemostatic fibrils.A kind of absorbable hemostatic fibrils were prepared by self-developed patent technique. The physical form and molecular structure of the fibrils and a marketed product Surgicel were characterized by general observation and infrared spectroscopy; the carboxyl content, pH value and relative molecular mass of fibrils were determined by potentiometric titration method, pH meter and copper ethylenediamine method, respectively. The behavior of the fibrils and Surgicel in contact with blood was observed by inverted microscope, the cytotoxicity was evaluated by agarose diffusion cell assay . The external iliac artery hemorrhage model and the back muscle infiltration model in rats were established. The hemostatic effectiveness of the fibrils was investigated by hemostasis time and blood weight, and the degradation and biosafety of fibrils were investigated by observation photography, immune organ weighing, hematology and coagulation index measuring, and histopathological examination. The fibrils and Surgicel had similar molecular structures. Compared with the raw material regenerated cellulose, the typical carboxyl stretching vibration absorption peak of -COOH appeared near in both fibrils and Surgicel. The carboxyl content of the two materials was about 20%, and the pH value was about 3. The relative molecular mass of the fibers after oxidation was 4466±79, which was close to that of Surgicel(>0.05). After contacting with blood, the volume of fibrils and Surgicel expanded, and absorbed blood of dozens of times as their own weight. The results of agar diffusion test showed that the fibrils had no cytotoxicity. The results of animal experiments showed that the hemostasis completed within and there was no significant difference in blood weight and speed of hemostasis between two products (both >0.05). The fibrils could be degraded 1 week after being implanted to the bleeding sites of the muscle. There were no pathological effects on the appearance, body weight, food intake, immunological tissue thymus, spleen, lymph nodes, hematology and coagulation indexes of the rats, and no obvious abnormality found in the histopathological examination. The prepared absorbable hemostatic fibrils have excellent biological safety and effectiveness.

Vegetable cellulose nanofiber dressing aids in the healing process of third-degree burns? study on rats / Curativo de nanofibras de celulose vegetal auxilia na cicatrização de queimaduras de terceiro grau? estudo em ratos

ABSTRACT Background: The treatment of 3rd degree burns represents a major medical challenge. Pinus vegetable cellulose is a biomaterial with characteristic similar to bacterial cellulose. Aim: To evaluate the safety of cellulose membrane (Pinus sp) in the treatment of 3rd burns in rats and to compare its effectiveness with the bacterial membrane already on the market. Method: Thirty-three Wistar rats were beaten with a 3rd degree burn on back skin by applying water at 98º C for 30 s. Then, they were divided into three groups (n=11): group 1 - simple dressing with gauze; group 2 - dressing with bacterial cellulose membrane; and group 3 - dressing with vegetable cellulose membrane. The animals were maintained for 15 days to check the general clinical status, macroscopic aspect, contraction of the wounds and microscopic analysis for the degree of healing and collagenization. Results: They were clinically well during the experiment. During the removal of the dressing, there was bleeding in the wound of the control group, unlike the groups treated with cellulose membranes, which protected the bed from injury. The macroscopic evaluation showed a greater contraction of the wounds treated with the membranes in relation to the control. A microscopic analysis revealed that most of the wounds were in advanced healing degree with predominance of mature collagen in all groups. Conclusion: Pinus sp cellulose membrane showed efficacy similar to that of the bacterial membrane in the treatment of 3rd degree burns.

RESUMO Racional: O tratamento das queimaduras de 3˚ grau representa grande desafio na área médica. A celulose vegetal de pinus é biomaterial com características semelhantes às da celulose bacteriana. Objetivo: Avaliar a segurança da membrana de celulose vegetal (Pinus sp) no tratamento de queimaduras de terceiro grau em ratos e comparar sua eficácia com a da membrana bacteriana já comercializada. Método: Trinta e três ratos Wistar foram submetidos à queimadura de 3º grau na pele do dorso mediante aplicação de água a 98º C durante 30 s. Em seguida, foram distribuídos em três grupos (n=11): grupo 1 - curativo simples com gaze; grupo 2 - curativo com membrana de celulose bacteriana; e grupo 3 - curativo com membrana de celulose vegetal . Os animais foram avaliados durante 15 dias para verificar o estado clínico geral, aspecto macroscópico, contração das feridas e análise microscópica pelo grau de cicatrização e colagenizacao. Resultados: Permaneceram clinicamente bem durante o experimento. Durante a retirada do curativo houve sangramento na ferida do grupo controle, diferentemente dos grupos tratados com as membranas de celulose, que protegeram o leito da lesão. A análise microscópica mostrou que a maioria das feridas apresentava-se em grau avançado de cicatrização, com predomínio de colágeno maduro em todos os grupos. Houve maior contração das feridas tratadas com as membranas em relação ao grupo controle. Conclus ão: A membrana de celulose de Pinus sp apresentou eficácia semelhante à da membrana bacteriana no tratamento de queimaduras de 3˚ grau.

Ocular surface findings in impression cytology after interferon a2b or mitomycin C in rabbits / Achados em citologia de impressão da superfície ocular após uso de interferon alfa-2b ou mitomicina C: estudo em coelho

ABSTRACT Objective: To describe ocular surface findings in impression cytology obtained from healthy rabbit conjunctiva treated with interferon alpha-2b eyedrop, and compare them to findings after use of mitomycin C 0.02%. Methods: An experimental study using a rabbit model was performed between September 2013 and October 2014 at the Faculdade de Medicina de Marília, Universidade Federal de São Paulo, Clínica de Olhos Moacir Cunha. Thirty New Zealand white rabbits were divided into 6 groups and received interferon alpha-2b or mitomycin C 0.02%. Impression cytology (IC) was performed prior to topical applications and at15, 30 and 60 days of use. The following variables were analyzed in impression cytology: goblet cells, cellularity, cell-to-cell adhesion, nucleus/cytoplasm ratio, nuclear chromatin, inflammatory cells keratinization, and cytomegaly. Results: The major findings in impression cytology after us of interferon alpha-2b included loss of goblet cells (50.8%), reduced cell-to-cell adhesion (26.2%), abnormal nucleus/cytoplasm ratio (20%) and reduced cellularity (15.4%). After use of mitomycin C 0.02%, the most common changes included loss of goblet cells (46.2%), abnormal nucleus/cytoplasm ratio (25.6%), less cell-to-cell adhesion (23.1%), and reduced cellularity (20.5%). There were no significant differences in any variable when comparing impression cytology after interferon alpha-2b and after mitomycin C 0.02%. Goblet cell loss was more pronounced at days 30 and 60, as compared to impression cytology at day 15 for both drugs. Conclusion: The loss of goblet cells, reduced cell-to-cell adhesion and cellularity, along with abnormal nucleus/cytoplasm ratio were the most common findings in impression cytology after use of interferon alpha-2b. These findings are similar to those described for use of mitomycin C 0.02%. ..

RESUMO Objetivo: Descrever os achados em citologia de impressão de conjuntiva sadia de coelho submetida ao uso de colírio de interferon alfa-2b e compará-los ao que foi encontrado após uso da mitomicina C 0,02%. Métodos: Estudo experimental realizado em modelo animal no período entre setembro de 2013 e outubro de 2014 nas dependências da Faculdade de Medicina de Marília, da Universidade Federal de São Paulo e da Clínica de Olhos Moacir Cunha. Trinta coelhos albinos da raça Nova Zelândia foram divididos em seis grupos e receberam interferon alfa-2b ou mitomicina C. A citologia de impressão foi realizada antes do início dos colírios e após 15, 30, 60 dias de seu uso. As seguintes variáveis foram analisadas na citologia de impressão: células caliciformes, celularidade, adesão intercelular, razão núcleo/citoplasma, cromatina, células inflamatórias, queratinização e citomegalia. Resultados: Os principais achados na citologia de impressão após o uso do interferon alfa-2b foram a redução de células caliciformes (50,8%), a diminuição da adesão intercelular (26,2%), a alteração da razão N/C (20%) e a redução da celularidade (15,4%). Após o uso da mitomicina C 0,02%, foram mais frequentes a redução das células caliciformes (46,2%), a alteração da razão N/C (25,6%), a adesão intercelular (23,1%) e a redução da celularidade (20,5%). Não houve diferença estatisticamente significante para nenhuma das variáveis estudas quando se compararam as citologias de impressão após interferon alfa-2b com as citologias de impressão após mitomicina C 0,02%. Independentemente da substância utilizada, as citologias colhidas 30 e 60 dias após início das drogas apresentaram maior redução de células caliciformes quando comparadas com as citologias de impressão colhidas após 15 dias. Conclusão: A redução das células caliciformes, a diminuição da adesão intercelular, a alteração da razão N/C e a diminuição da celularidade foram as alterações mais frequentes na citologia de impressão colhida após o uso de interferon alfa-2b. Os achados em citologias de impressão após o uso de interferon alfa-2b são semelhantes àqueles encontrados após o uso da mitomicina C 0,02%.

Desenvolvimento de máscaras de tecido e celulose para resposta emergencial à pandemia provocada pelo SARS-CoV-2 / Development of tissue and cellulose masks for emergency response to the SARS-CoV-2 pandemic / Desarrollo de máscaras de tejido y celulosa para la respuesta de emergencia a la pandemia de SARS-CoV-2

Objetivo: desenvolver um modelo de máscara de tecido, com aplicação do elemento filtrante em celulose, para fins de utilização como barreira física segura para aerossóis, como estratégia de resposta emergencial à pandemia provocada pelo SARS-CoV-2. Método: pesquisa laboratorial realizada por meio de protótipos, testagens empíricas e análises e discussões junto a expertises. Resultados: a condução da pesquisa demonstrou que os aerossóis são retidos pela barreira física de celulose introduzida à estrutura das máscaras, o que motivou a segunda fase do estudo em unidade da Rede Brasileira de Laboratórios Analíticos de Saúde sobre a eficácia desses materiais. Conclusão: a confecção de máscaras de tecido é um fenômeno mundial importante e urgente frente à pandemia da COVID-19. Em função da crise de abastecimento e dos parâmetros ressaltados neste estudo, acredita-se que o uso desse equipamento possa ser estendido a setores não críticos de unidades de saúde, além da população em geral.

Objective: to develop a model of fabric mask, with the application of a cellulose filter element, for use as a safe physical barrier for aerosols, as an emergency response strategy for the SARS-CoV-2 pandemic. Method: laboratory research carried out by means of prototypes, empirical tests and analyses, and discussions with experts. Results: the research demonstrated that aerosols are retained by the physical cellulose barrier introduced into the structure of the masks, which motivated the second phase of the study into the effectiveness of these materials at a unit of the Brazilian Analytical Health Laboratories Network. Conclusion: the making of fabric masks is an important and urgent worldwide phenomenon in tackling the COVID-19 pandemic. In view of the supply crisis and the parameters highlighted in this study, it is believed that the use of this equipment can be extended to non-critical sectors of health units, as well as to the general population.

Objetivo: desarrollar un modelo de mascarilla de tela, con la aplicación de un elemento filtrante de celulosa, para su uso como barrera física segura para aerosoles, como estrategia de respuesta de emergencia para la pandemia SARS-CoV-2. Método: investigación de laboratorio realizada mediante prototipos, pruebas y análisis empíricos y discusiones con expertos. Resultados: la investigación demostró que los aerosoles son retenidos por la barrera física de celulosa introducida en la estructura de las máscaras, lo que motivó la segunda fase del estudio sobre la efectividad de estos materiales en una unidad de la Red Brasileña de Laboratorios Analíticos de Salud. Conclusión: la fabricación de máscaras de tela es un fenómeno mundial importante y urgente para hacer frente a la pandemia de COVID-19. Ante la crisis de oferta y los parámetros resaltados en este estudio, se cree que el uso de este equipamiento puede extenderse a sectores no críticos de las unidades de salud, así como a la población en general.

Traumatic perforations of the tympanic membrane: immediate clinical recovery with the use of bacterial cellulose film / Perfurações traumáticas da membrana timpânica: recuperação clínica imediata com o uso de fragmento de celulose bacteriana

Abstract Introduction: Perforation of the tympanic membrane is a reasonably frequent diagnosis in otorhinolaryngologists' offices. The expectant management is to wait for spontaneous healing, which usually occurs in almost all cases in a few weeks. However, while waiting for healing to be completed, the patients may experience uncomfortable symptoms. Although some research suggests the use of various materials to aid in the recovery of the tympanic membrane, none presented robust evidence of improvement in the cicatricial process. Nevertheless, the occlusion of the perforation with some material of specific texture and resistance can alleviate the patients' symptoms and accelerate the healing process. Objective: To evaluate the clinical (symptomatic and functional) improvement after the placement of bacterial cellulose film (Bionext®) on tympanic membrane perforations (traumatic). Methods: We evaluated 24 patients, victims of traumatic perforations of the tympanic membrane, who were evaluated in the Otorhinolaryngology Emergency Room. Following otoscopy and audiometric examination was performed, before and after the use of cellulose film occluding the tympanic membrane perforation. Results: Twenty-four patients were included, whose degree of overall discomfort caused by the tympanic membrane perforation and the presence of symptoms of autophonia, ear fullness and tinnitus were investigated. The mean score attributed to the overall annoyance caused by tympanic membrane perforation was 7.79, decreasing to a mean value of 2.25 after the film application. Symptom evaluation also showed improvement after using the film: autophonia decreased from a mean value of 6.25 to 2.08, tinnitus from 7 to 1.92 and ear fullness from 7.29 to 1.96. The auditory analysis showed mean threshold values still within the normal range at low and medium frequencies, with slight hearing loss at acute frequencies, but with significant improvement at all frequencies, with the exception of 8000 Hz, after film use. Conclusion: The use of bacterial cellulose film fragment on traumatic perforations of the tympanic membrane promoted immediate functional and symptomatic recovery in the assessed patients.

Resumo Introdução: A perfuração da membrana timpânica é uma condição clínica relativamente frequente em consultórios de otorrinolaringologistas. A conduta é quase sempre expectante, aguardando cicatrização espontânea, que costuma ocorrer na quase totalidade dos casos em algumas semanas. No entanto, enquanto não se completa, os pacientes mantêm sintomas desconfortáveis. Embora algumas pesquisas sugiram o uso de materiais diversos para auxiliar na recuperação da membrana timpânica, nenhuma apresentou evidência substancial de melhoria no processo cicatricial. Por outro lado, a oclusão da perfuração com alguns materiais de textura e resistência específicas poderia aliviar os sintomas dos pacientes durante o processo cicatricial. Objetivo: Avaliar a melhoria clínica (sintomática e funcional) após a colocação de película de celulose bacteriana (Bionext®) sobre a perfuração da membrana timpânica (traumática). Método: Foram avaliados pacientes com perfurações traumáticas da membrana timpânica que deram entrada no pronto-socorro de otorrinolaringologia. Eles foram avaliados em relação a suas queixas e exame audiométrico, antes e após a aplicação de película de celulose que oclui a perfuração da membrana timpânica. Resultados: Foram incluídos 24 pacientes, nos quais foram pesquisados o grau de incômodo global causado pela perfuração da membrana timpânica e a presença de sintomas de autofonia, plenitude auricular e tinnitus. A média da nota atribuída ao incômodo global causado pela perfuração foi de 7,79; caiu para valor médio de 2,25 após a aplicação da película. A avaliação dos sintomas também apresentou melhoria após uso da película, autofonia caiu de valor médio de 6,25 para 2,08; zumbido de 7 para 1,92 e plenitude auricular de 7,29 para 1,96. A análise auditiva apresentou um valor médio de limiares ainda dentro da normalidade em frequências baixas e médias, com perda de audição leve em frequências agudas, porém com melhoria significante em todas as frequências, com exceção de 8000 Hz, após a colocação da película. Conclusão: A aplicação de fragmento de película de celulose bacteriana sobre perfurações traumáticas da membrana timpânica promoveu recuperação funcional e sintomática imediata nos pacientes estudados.

Moringa straw as cellulase production inducer and cellulolytic fungi source / Residuo de moringa como inductor de la producción de celulasas y como fuente de hongos celulolíticos

Abstract Currently, the valorization of agroindustrial waste is of great interest. Moringa oleifera is a multipurpose tree whose softwood residues could be used as raw material for low-cost cellulase production. The aim of this study was to isolate, identify, and characterize microorganisms with cellulolytic activity in different carbon sources. We isolated and puri-fied 42 microorganisms from M. oleifera biomass. Fungi presenting the largest hydrolytic halos in carboxymethylcellulose as a substrate were molecularly identified as Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) and Cladosporium cladosporioides (FC2). The ability of these fungal strains to break down cellulose was assessed in a submerged fermentation using either amorphous CMC or crystalline form (Avicel). P. funiculosum and C. cladosporioides displayed similar endoglucanase (606 U/l) and exoglucanase (205 U/l) activities in the Avicel-containing medium, whereas F. verticillioides showed the highest level of p-glucosidase activity (664 U/l) in the carboxymethylcellulose medium. In addition, the effect of three culture media (A, B, and C) on cellulase production was evaluated in P. funiculosum using moringa straw as a carbon source. The results showed a volumetric productivity improvement of cellulases that was 2.77-, 8.26-, and 2.30-fold higher for endoglucanase, exoglucanase and p-glucosidase, respectively when medium C containing moringa straw was used as a carbon source. The enzymatic extracts produced by these fungi have biotechnological potential especially for second-generation bioethanol production (2G) from moringa straw. This is the first report on the use of M. oleifera biomass to induce the production of various cellulases in P. funiculosum. © 2019 Asociación Argentina de Microbiología. Published by Elsevier Espana, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Resumen Actualmente, la valorización de los residuos agroindustriales es de gran interés. En este trabajo se emplearon residuos de madera blanda de Moringa oleifera para la producción de celulasas de bajo costo. El objetivo fue aislar, identificar y caracterizar microorganismos con actividad celulolítica en diferentes fuentes de carbono. A partir de la biomasa de M. oleifera, se aislaron e identificaron 42 microorganismos productores de celulasas. Los hongos que presentaron los mayores halos de hidrólisis en carboximetilcelulosa como sustrato fueron identificados molecularmente como Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) y Cladosporium cladosporioides (FC2). Mediante fermentación sumergida, se evaluó la capacidad de estas cepas en la producción de celulasas utilizando celulosa cristalina (Avicel) y amorfa (CMC) como fuentes de carbono. P. funiculosum y C. cladosporioides presentaron las mayores actividades de endoglucanasa (606 U/l) y exoglucanasa (205 U/l) en medio Avicel, mientras que F. verticillioides mostró la mayor actividad de p-glucosidasa (664 U/l) en medio CMC. Además, se evaluó el efecto de tres medios de cultivo (A, B y C) sobre la producción de celulasas en P. funiculosum empleando residuos de moringa como fuente de carbono. Los resultados mostraron que en el medio C, la productividad volumétrica de celulasas se incrementó en 2,77; 8,26 y 2,30 veces para las actividades de endoglucanasa, exoglucanasa y p-glucosidasa, respectivamente. Los extractos enzimáticos producidos tienen gran potencial para su utilización biotecnológica, especialmente en la sacarificación de residuos de moringa y la producción de bioetanol de segunda generación. Este es el primer estudio del uso de la biomasa de M. oleifera para inducir la producción de diversas celulasas en P. funiculosum.

Comparative study on infection and degradation of Armillaria gallica and Phallus impudicus to fungus-growing materials / 中国中药杂志

The phenomenon that waste of fungus-growing materials in the planting process of Gastrodia elata is very common. It has been proved by practice that the used fungus-growing materials planted with G. elata can be used to plant Phallus impudicus. But the mechanism is unclear. In this study, we compared the different infested-capacity of Armillaria gallica and Phallus impudicus by morphological anatomy of the used fungus-growing materials. We also compared the differences on the two fungi consumed the main contents of fungus-growing materials, cellulose, lignin and hemicellulose, by using nitric acid-95% ethanol method, sulfuric acid method and tetrabromide method respectively, so that to explore the mechanism of A. gallica and P. impudicus recycle the fungus-growing materials, and to provide scientific basis for recycling the used fungus-growing materials of G. elata. The results showed that A. gallica had a strong ability to invade some parts outside the vascular cambium, but it had a weak ability to invade some parts inside the vascular cambium, while P. impudicus had a strong ability to invade the same parts. The contents of lignin and cellulose, which from inside and outside the vascular cambium of fungus-growing materials were significantly different. In the parts of outside the vascular cambium of fungus-growing materials, A. gallica degraded more lignin and cellulose, while P. impudicus degraded more hemicellulose. In the parts of inside the vascular cambium of fungus-growing materials, A. gallica degraded more cellulose, while P. impudicus degraded more hemicellulose. The present results suggested that A. gallica and P. impudicus made differential utilization of the carbon source in the fungus-growing materials to realize that P. impudicus recycle the used fungus-growing materials of G. elata. A. gallica used lignin and cellulose as the main carbon source, while P. impudicus used hemicellulose as the main carbon source.

Pine nanocellulose and bacterial nanocellulose dressings are similar in the treatment of second-degree burn? experimental study in rats / Os curativos de nanocelulose de pinus e nanocelulose bacteriana são semelhantes no tratamento da queimadura de segundo grau? estudo experimental em ratos

ABSTRACT Background: Despite all the advances in medicine and the wide variety of dressings available, the treatment of burn wounds still represents an important medical challenge. The pinus cellulose membrane dressing is a biomaterial with characteristics similar to those of bacterial cellulose, but with lower cost. Aim: To evaluate the efficacy of pinus nanocellulose membrane on healing of deep second degree burns in rats and compare with Membracel®. Method: Thirty male Wistar rats were submitted to deep second degree burn in dorse, with boiling water at 97o C for 20 s, generating a 314 mm² area wound. The animals were distributed in three dressing groups (n=10): group 1 - simple gauze; group 2 - bacterial cellulose membrane (Membracel®); and group 3 - pinus cellulose membrane. They were evaluated for 20 days to verify clinical condition, macro and microscopic appearance and wound contraction. Results: All of them remained clinically well with no differences in weight. Crusts were observed in group 1, and none in groups 2 and 3. Regarding to scar contraction, groups 2 and 3 were similar, better than group 1. Microscopic analysis showed predominance of advanced healing degree in groups 1 and 3, and initial in group 2. Mature collagen was predominant in all groups. Conclusion: The pinus nanocellulose membrane is effective in the treatment of experimental second degree burn in rats and its effectiveness is similar to that of the bacterial nanocellular membrane.

RESUMO Racional: Apesar de todos os avanços da medicina e da grande variedade de curativos disponíveis, o tratamento das queimaduras ainda representa importante desafio médico. O curativo de membrana de celulose de pinus é biomaterial com características semelhantes à de celulose bacteriana, mas de menor custo. Objetivo: Avaliar a eficácia da membrana de nanocelulose de pinus na cicatrização de queimaduras profundas de segundo grau em ratos e comparar com a Membracel®. Método: Trinta ratos Wistar machos foram submetidos à queimadura profunda de segundo grau em dorso, com água fervente a 97o C por 20 s, gerando lesão de 314 mm². Os animais foram distribuídos em três grupos de curativos (n = 10): grupo 1 - gaze simples; grupo 2 - membrana de celulose bacteriana (Membracel®); e grupo 3 - membrana de celulose de pinus. Eles foram avaliados por 20 dias para verificar o quadro clínico, aspecto macro e microscópico e a contração da ferida. Resultados: Todos permaneceram clinicamente bem, sem diferenças de peso. Crostas foram observadas no grupo 1 e nenhuma nos grupos 2 e 3. Em relação à contração da cicatriz, os grupos 2 e 3 foram semelhantes, melhores que o grupo 1. A análise microscópica mostrou predomínio de grau de cicatrização avançado nos grupos 1 e 3, e inicial no grupo 2. O colágeno maduro foi predominante em todos os grupos. Conclusão: A membrana de nanocelulose de pinus é eficaz no tratamento de queimaduras experimentais de segundo grau em ratos e sua eficácia é semelhante à da membrana nanocelular bacteriana.

Micro and nanocrystalline cellulose based oral dispersible film; preparation and evaluation of in vitro/in vivo rapid release studies for donepezil

Oral fast-dispersible film was prepared by utlizing donepezil hydrochloride (drug) and various cellulose derivatives such as hydroxypropyl methyl cellulose (hypermellose) (HPMC), microcrystalline cellulose (MCC) and nanocrystalline cellulose (NCC) to treat Alzheimer's disease. NCC was synthesized by ultra-sonication method using MCC and this was converted to thinfilm formulation (NCC-F) using solvent casting technique. The interaction between the polymer and the drug was investigated by spectral analysis such as UV, FTIR, and 1H- NMR. FTIR confirmed that the compatibility of drug and polymer in ODF formulation. NCC-F has shown an average surface roughness of 77.04 nm from AFM and the average particle size of 300 nm from SEM analysis. Nano sized particle of NCC-F leads faster in vitro dissolution rate (94.53%) when compared with MCC-F and F3 formulation. Animal model (in vivo) studies of NCC-F formulation has reached peak plasma concentration (Cmax) up to 19.018 ng/mL in the span of (tmax) 4 h with greater relative bioavailability of 143.1%. These results suggested that high surface roughness with nanosized NCC-F formulation attained extended drug availability up to (t1/2) 70 h.