Desenvolvimento e caracterização de lipossomas peguilados para veiculação de L-asparaginase / Development and characterization of peg-grafted liposomes for lasparaginase encapsulation

A Leucemia Linfoide Aguda (LLA) é um câncer de maior incidência em crianças, e tem a Lasparaginase (ASNase) como fármaco amplamente utilizado no tratamento dos afetados. A ASNase catalisa a hidrólise do aminoácido L-asparagina (Asn), presente na corrente sanguínea, a ausência do aminoácido no meio extracelular leva à morte células leucêmicas, que necessitam deste aminoácido para as funções celulares. Fatores envolvendo a eficiência do tratamento com ASNase como reações adversas e curta meia-vida, principalmente devido ao reconhecimento pelo sistema imune e degradação por proteases, limitam a sua eficácia. A encapsulação da enzima em lipossomas pode conferir proteção à degradação, melhorar seu perfil farmacocinético e diminuir os efeitos adversos, de forma a melhorar o tratamento da LLA sendo este o objetivo desse trabalho. Lipossomas de DOPC (1,2-dioleoil-sn-glicero-3-fosfocolina) e DMPC (1,2-dimiristoil-snglicero-3-fosfocolina) foram desenvolvidos empregando-se o método de hidratação do filme lipídico e diferentes protocolos de preparo contendo ou não diferentes concentrações de 18:0 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polietilenogicol)-2000] (DSPE-PEG). Os lipossomas produzidos foram utilizados para encapsular a ASNase e os sistemas contendo ou não ASNase encapsulada foram caracterizados por espalhamento de luz dinâmico (DLS), potencial zeta, microscopia eletrônica de transmissão (MET) e criomicroscopia de transmissão. Adicionalmente, foram avaliados a taxa de encapsulação e o perfil de permeabilidade das vesículas à L-asparagina. As análises de DLS mostraram que as nanoestruturas formadas empregando-se agitação magnética a partir de sistemas contendo 10% e 20% de DSPE-PEG possuem diâmetro hidrodinâmico menor (~ 25 nm a 60 nm) que os mesmos sistemas sem o fosfolipídio peguilado (~190 nm a 222 nm), demonstrando a relação entre a diminuição do tamanho e o aumento da quantidade de fosfolipídio peguilado e possível formação de estruturas micelares ou bicelares. O emprego de agitação em vórtex para hidratação do filme lipídico, adição do antioxidante -tocoferol e redução da concentração de DSPE-PEG (5% e 10%) levou à formação de sistemas com diâmetro hidrodinâmico maior, sendo esse protocolo e concentrações de PEG definidos como padrão. As análises de MET comprovaram a formação de lipossomas com diâmetro hidrodinâmico semelhante ao observado por DLS; com a utilização da criomicroscopia foi possível observar os lipossomas sem deformações. Os lipossomas de DMPC/DSPE-PEG 10% apresentaram maior permeabilidade à L-asparagina ao longo do tempo e, portanto, poderiam funcionar como nanoreatores, depletando o aminoácido da circulação. Estudos in vitro com células tumorais devem ser realizados e em seguida estudos in vivo, para confirmar este potencial

L-asparaginase (ASNase) is a first-choice drug, combined with other drugs, in therapeutic schemes to treat Acute Lymphoblastic Leukemia (ALL) in children and adolescents. ASNase catalyzes the hydrolysis of L-asparagine (Asn) in the bloodstream; since ALL cells cannot synthesize this amino acid, protein synthesis is impaired leading to leukemic cells death by apoptosis. In spite of its therapeutic importance, treatment with ASNase is associated to side effects, mainly hypersensitivity and immunogenicity. Another drawback refers to degradation by plasma proteases that altogether with immunogenicity shortens the enzyme half-life. Encapsulation of ASNase in liposomes, vesicular nanostructures formed by the self-aggregation of phospholipids, is an attractive alternative that possibly will protect the enzyme from plasma proteases, resulting on better pharmacokinetics profile. In this work, we prepared by thin film hydration liposomal formulations of the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dimyristoyl-sn-glycero-3- phosphocholine (DMPC) containing or not different concentrations of 18:0 1,2-distearoyl-snglycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG), and encapsulated ASNase by electroporation. The systems containing or not ASNase were analyzed by Dynamic Light Scattering, zeta potential and Electron Microscopy. The encapsulation efficiency and vesicles permeability were also evaluated. According to the DLS analysis, the nanostructures formed by film hydration under magnetic stirring employing 10% or 20% DSPE-PEG presented smaller hydrodynamic diameter (~ 25 nm to 60 nm) than the same systems without the pegylated phospholipid (~ 190 nm to 222 nm), demonstrating the relation between size and the amount of pegylated phospholipid that results in formation of micellar or bicellar structures. The protocol was stabilize by hydration of the lipid film under vortex agitation, addition of the antioxidant - tocopherol and reduction of the concentration of DSPE-PEG (5% and 10%), what altogether led to the formation of nanostructures of higher hydrodynamic diameter and monodisperse systems. TEM analyzes confirmed the formation of liposomes with hydrodynamic diameter similar to that observed by DLS; with the use of cryomicroscopy it was possible to observe the liposomes without deformations. Liposomes of DMPC/DSPE-PEG 10% showed permeability to L-asparagine over time and, therefore, could function as nanoreactors, depleting the circulating amino acid

Hallazgos ultraestructurales renales de pacientes con nefritis intersticial crónica de las comunidades agrícolas en El Salvador / Renal ultrastructural findings of patients with chronic interstitial nephritis from farming communities in El Salvador

Introducción: Desde hace 20 años se presenta en Centroamérica una enfermedad renal crónica que fundamentalmente afecta a hombres agricultores y no asociada a las causas tradicionales. Se caracteriza por presentar una nefritis intersticial crónica, en tanto las características ultraestructurales no se conocen con exactitud. En su origen se invoca el uso de agroquímicos y otros agentes nefrotóxicos, la deshidratación crónica, el consumo de medicamentos, entre otros factores. Objetivo: Describir las características ultraestructurales de la nefritis intersticial crónica en comunidades agrícolas. Método: Se realizó un estudio descriptivo de corte transversal. Se estudiaron muestras de biopsias renales de ocho pacientes con diagnóstico de nefritis intersticial crónica de las comunidades agrícolas. Resultados: De los ocho pacientes estudiados, dos (25 por ciento) trabajaban en labores agrícolas y cinco eran del sexo femenino (62,5 por ciento). Dos de los pacientes (25 por ciento) presentaban una enfermedad renal crónica estadio 2, y seis (75 por ciento) estadio 3. En cinco pacientes se hallaron fagolisosomas con presencia de componente lipídico entremezclado con material electrodenso en células del túbulo distal. En igual cantidad de pacientes se observaron cuerpos mieloides con zonas laminadas y núcleo central en células de túbulo proximal y de los vasos sanguíneos. Conclusiones: En pacientes de comunidades agrícolas que padecen nefritis intersticial crónica se evidencian fagolisosomas y estructuras mieloides en túbulos y vasos renales, cuyo contenido y origen se desconocen(AU)

Introduction: Chronic kidney disease mainly affecting male farmers and not associated to traditional causes has been present in Central America for twenty years. The condition is characterized by the presence of chronic interstitial nephritis, but its ultrastructural features are not fully known. Factors suggested as responsible for its occurrence include the use of agrochemicals and other nephrotoxic agents, chronic dehydration and medicine consumption. Objective: Describe the ultrastructural characteristics of chronic interstitial nephritis in farming communities. Method: A cross-sectional descriptive study was conducted of renal biopsy samples from eight patients diagnosed with chronic interstitial nephritis in farming communities. Results: Of the eight patients studied, two (25 percent) were farm workers and five (62.5percent) were female. Two of the patients (25 percent) had stage 2 and six (75 percent) stage 3 chronic kidney disease. In five patients evidence was found of phagolysosomes with lipid component mixed with electrodense material in distal tubule cells. An equal number of patients had myeloid bodies with laminated areas and central nucleus in proximal tubule and blood vessel cells. Conclusions: Evidence of phagolysosomes and myeloid structures of unknown content and origin was found in renal tubules and vessels of patients from farming communities diagnosed with chronic interstitial nephritis(AU)

Morphology and infection process of Olivea neotectonae in teak leaves / Morfologia e processo de infecção de Olivea neotectonae em folhas de teca

Rust, caused by the biotrophic fungus Olivea neotectonae, stands out as an important disease in teak plantations in Brazil. Considering the current importance of this rust and the need to obtain additional basic information about its causal agent to better disease management, the present study aimed to analyze the morphology of O. neotectonae using light microscopy and demonstrate the infection process in teak leaves using scanning electron microscopy techniques (SEM). The morphological characteristics observed using light microscopy were similar to those described in the literature for this species. SEM revealed that the infection structures of the pathogen penetrated the teak leaf through the stomata complex. This study provides the first microscopic evidence of the infection process of Olivea neotectonae in leaves of teak plants.(AU)

A ferrugem, causada pelo fungo biotrófico Olivea neotectonae, destaca-se como uma doença importante nas plantações de teca no Brasil. Considerando a importância atual dessa ferrugem e a necessidade de obter informações básicas adicionais sobre seu agente causal para melhor manejo da doença, o presente estudo teve como objetivo analisar a morfologia da O. neotectonae utilizando microscopia de luz e demonstrar o processo de infecção em folhas de teca por meio de técnicas de microscopia eletrônica de varredura (MEV). As características morfológicas observadas em microscopia de luz foram semelhantes às descritas na literatura para essa espécie. A MEV revelou que as estruturas de infecção do patógeno penetraram na folha de teca através do complexo estomático. Este estudo fornece a primeira evidência microscópica do processo de infecção de Olivea neotectonae em folhas de plantas de teca.(AU)

Biosynthesized silver nanoparticles: are they effective antimicrobials?

BACKGROUND Silver nanoparticles (AgNPs) are increasingly being used in medical applications. Therefore, cost effective and green methods for generating AgNPs are required. OBJECTIVES This study aimed towards the biosynthesis, characterisation, and determination of antimicrobial activity of AgNPs produced using Pseudomonas aeruginosa ATCC 27853. METHODS Culture conditions (AgNO3 concentration, pH, and incubation temperature and time) were optimized to achieve maximum AgNP production. The characterisation of AgNPs and their stability were evaluated by UV-visible spectrophotometry and scanning electron microscopy. FINDINGS The characteristic UV-visible absorbance peak was observed in the 420-430 nm range. Most of the particles were spherical in shape within a size range of 33-300 nm. The biosynthesized AgNPs exhibited higher stability than that exhibited by chemically synthesized AgNPs in the presence of electrolytes. The biosynthesized AgNPs exhibited antimicrobial activity against Escherichia coli, P. aeruginosa, Salmonella typhimurium, Staphylococcus aureus, methicillin-resistant S. aureus, Acinetobacter baumannii, and Candida albicans. MAIN CONCLUSION As compared to the tested Gram-negative bacteria, Gram-positive bacteria required higher contact time to achieve 100% reduction of colony forming units when treated with biosynthesized AgNPs produced using P. aeruginosa.

Comparative study on morpho-anatomy of leaf, stem and root of Boerhaavia diffusa L. (Nyctaginaceae) and its adulterant plants

ABSTRACT Punarnava (Boerhaavia diffusa L.- Nyctaginaceae) is a promising drug to rejuvenate new cells in the body. It is well known in Ayurvedic medicine and locally called Tambadivasu. Superficially it is similar to other species of Boerhaavia and species of Trianthema and Sesuvium. Due to the minute morphological differences, the above plants are erroneously used in medicine as Punarnava, and at times on purpose as an adulterant. Therefore, it is necessary to highlight the anatomical features of Punarnava for proper identification of the medicinal plant species for local people and for scientific research. Due to the ambiguity in local names and similar apparent appearance, market samples of Punarnava are often adulterated with various species of Trianthema and Sesuvium. These adulterated samples contain neither the Punarnavine alkaloid, nor does it possess anisocytic stomata but possess paracytic stomata. Comparative study of stem anatomy showed two main characteristic differences. First, plenty of starch grains can be seen in both the ground parenchymatous tissues present in between successive cambia and xylem parenchyma of Punarnava which is not observed in species of Trianthema, and second, the phloem around the xylem of Punarnava root has semi-circular or eccentric patches, while that of Trianthema only has narrow strips. This study is focused on comparative SEM study of leaf morphologies and anatomy of leaf, stem, and root of Boerhaavia diffusa L., Trianthema portulacastrum L. and Sesuvium portulacastrum L.

Análise do papel dos exossomos derivados das linhagens celulares SCC-15 e HSC-3 no processo de angiogênese tumoral / Analysis of the role of exosomes derived from cell lines SCC-15 and HSC-3 in the process of tumor angiogenesis

Os exossomos são vesículas extracelulares originadas por brotamento interno da membrana de endossomos tardios que representam uma eficiente forma de comunicação intercelular. Devido às suas múltiplas funções biológicas, o foco de alguns estudos atuais tem se concentrado na análise do seu papel no desenvolvimento do câncer, progressão da doença, invasão, angiogênese e formação de metástases tumorais. Nesta perspectiva, o presente estudo objetivou caracterizar os exossomos secretados por duas linhagens celulares de carcinomas epidermoide oral (CEO) (SCC-15 e HSC-3) e avaliar seus efeitos sobre uma linhagem de células endoteliais (HUVEC), em relação à sua capacidade de formação de estruturas vasculares, taxas de migração, proliferação e índices de apoptose/necrose. Médias significativamente maiores de células com potencial invasivo (p<0,0001) e migratório (p<0,0001) foram observadas para a linhagem HSC-3. Ultraestruturalmente, verificou-se que as partículas derivadas da linhagem SCC-15 exibiram morfologia arredondada e diâmetro inferior a 150 nm. Nenhuma diferença estatisticamente significativa foi revelada entre as linhagens celulares estudadas, considerando a quantificação de nanovesículas (p=0,2252) e tamanho exossomal (p=0,1765). Por imunofluorescência indireta, identificou-se que 22,15% dos exossomos secretados pelas células SCC-15 e 18,37% dos exossomos derivados da linhagem HSC-3 expressaram o anticorpo anti-Anexina. No que se refere aos ensaios funcionais com as HUVECs, o tratamento com exossomos derivados da linhagem SCC-15 induziu um aumento significativo da capacidade de formação de estruturas vasculares (p<0,0001), potencial migratório (p=0,0016) e taxa de apoptose (p<0,0001), enquanto que uma redução da proliferação celular foi apontada (p=0,0030). Por outro lado, o tratamento com exossomos secretados pela linhagem HSC-3 promoveu uma redução significativa da formação tubular (p<0,0001), motilidade (p=0,0042) e proliferação celular (p=0,0010), ao passo que nenhuma diferença estatisticamente significativa foi observada no índice apoptótico (p=0,3004). Os resultados do presente estudo indicaram a participação dos exossomos derivados de linhagens de CEO no processo de angiogênese tumoral, onde as células SCC-15 exibiram forte resposta proangiogênica e a linhagem HSC-3 demonstrou efeito antiangiogênico (AU).

Exosome are extracellular microvesicles originated by inward budding of late endosomal membrane that represent an efficient form of intercellular communication. Because of its multiple biological functions, the focus of some recent studies has concentrated on the analysis of its role in cancer development, disease progression, invasion, angiogenesis and tumor metastasis formation. In this perspective, the present study aimed to characterize the secreted exosomes by two cell lines of oral squamous cell carcinomas (OSCC) (SCC-15 and HSC-3) and to evaluate its effects on a cell line of endothelial cells (HUVEC), in relation to their ability to form vascular structures, rates of migration, proliferation, and apoptosis / necrosis indices. Significantly higher means of cells with invasive (p<0.0001) and migratory potential (p = <0.0001) were observed for the HSC-3 cell line. Ultrastructurally, it was found that particles derived from the SCC-15 cell line exhibited a rounded morphology and diameter of less than 150 nm. No statistically significant difference was revealed between the studied cell lines, considering the nanovesicles quantization (p=0.2252) and exossomal size (p=0.1765). By indirect immunofluorescence, it was found that 22.15% of exosomes secreted by SCC-15 cells and 18.37% of exosomes derived from HSC-3 cells expressed anti-annexin antibody. With regard to the functional tests with HUVECs, treatment with exosomes derived from SCC-15 cell line induced a significant increase in their capacity of formation of vascular structures (p = <0.0001), migratory potential (p=0.0016) and rate of apoptosis (p<0.0001), while a decrease in cell proliferation was noted (p = 0.0030). On the other hand, the treatment with exosomes secreted by HSC-3 cell line produced a significant reduction in tubule formation (p<0.0001), motility (p = 0.0042) and cell proliferation (p=0.0010), whereas no statistically significant difference was observed in the apoptotic index (p=0.3004). The results of this study indicated the involvement of exosomes derived from OSCC cell lines in tumor angiogenesis process, in which the SCC-15 cells exhibited strong proangiogenic response and HSC-3 cell line showed antiangiogenic effect (AU).

Aplicação da avaliação ultraestrutural de espermatozoides na rotina da andrologia / Application of sperm ultra structural evaluation in andrology routine

A microscopia eletrônica de espermatozoides é uma ferramenta complementar da análise seminal que pode contribuir na interpretação clínica da astenozoospermia grave e da teratozoospermia e na investigação de infertilidade idiopática. Reportamos um caso de paciente com varicocele, submetido à varicocelectomia, com análise seminal ultraestrutural por microscopia eletrônica.

Electron microscopy of sperm is a complementary tool to semen analysis that can contribute to the clinical interpretation of severe astenozoospermia, teratozoospermia and idiopathic infertility investigation. We report a patient with varicocele, submitted to varicocelectomy,with seminal ultrastructural analysis by electron microscopy.

Diarreia aguda / Acute diarrhea

Diarreia aguda é a passagem de quantidade acima do normal de fezes amolecidas associada ao aumento do número de evacuações. No diagnóstico diferencial das diarreias agudas devem ser enfocados as infecções, as alergias alimentares, a intoxicação alimentar, o uso de medicações e a apresentação inicial de diarreia crônica. Dentre estas possíveis etiologias, especialmente em nosso meio, as causas infecciosas devem sempre vir à mente e constituir uma das primeiras opções na investigação diagnóstica. As infecções intestinais associadas a quadros diarreicos são a segunda causa de mortes de origem infecciosa em todo o mundo, com prevalência estimada de 3 a 5 bilhões de casos/ano. Os autores atualizam as novidades e peculiaridades a respeito do diagnóstico e dos tratamentos — geral e/ou específico — dos diferentes agentes associados à diarreia aguda infecciosa.

Acute diarrhea is the passage of above normal quantities of soft faeces also associated with increased bowel movements. Differential diagnosis of acute diarrhea should be focused on infections, food allergies, food poisoning, use of medications and the initial presentation of chronic diarrhea. Among these possible etiologies, given the environment we live in, infectious causes should always be taken into account and be one of the first options in diagnostic investigation. Intestinal infections associated with diarrheal frames are the second leading cause of infectious deaths worldwide, with an estimated to 3-5 billion cases/per year. In this review, the authors intend to review the new features and aspects concerning diagnosis and treatment — general and/or specific — of the different agents associated with acute infectious diarrhea.

Morphological study of bone cranial in athymic mice / Estudio morfológico de hueso craneal en ratones atímicos

The aim of our research was to create an osteogenic unit in the skulls of athymic mice; however, the first challenge we faced was to find sufficient and adequate data that would allow us to determine the morphological, immunohistochemical and microtopographical characteristics that could be used as normality standards in athymic mice skulls and, hence, a reference in the event of achieving the formation of de novo bone using the osteogenic unit we proposed. Knowing the normal bone morphology in the skull of athymic mice was a necessary precondition to develop subsequently an osteogenic unit possessing the Osteogenesis, Osteoinduction and Osteoconductivity that could be compared versus those in the normal bone during its formations and remodeling processes. Therefore, we conducted a pilot study to determine bone morphological characteristics in the skull of athymic mice by means of specific histological staining: hematoxylin-eosin and Von Kossa, for osteoid tissue and mineralized bone, and Masson Tri-chrome for ossified areas. We also use immunohistochemistry to detect bone formation markers: alkaline phosphatase resulting from osteoblastic activity stimulation, type 1 collagen a bonematrix structural protein; Osteopontine, a protein specifically synthesized by osteoblasts that favors cell proliferation and remodeling in bone defects; Osteocalcine, a peptide hormone produced by osteoblasts during bone formation; and, Runx 2, a transcription factor expressed by stem cells which stimulates bone differentiation. Likewise, we used electron microscopy on the newly formed tissue to determine the presence of organic deposits, such as calcium, phosphate and magnesium in bone tissue.

Propusimos la realización de una unidad osteogénica a desarrollar en cráneo de ratones atímicos, Sin embargo, nos enfrentamos al reto de encontrar datos que nos determinaran cuales eran las características morfológicas, inmunohistoquímicas y micro-topográficas del cráneo de estos ratones atímicos, que nos sirvieran como referencia de normalidad y tener un punto de comparación, en caso de que pudiéramos lograr la formación de hueso de novo, a partir de la unidad osteogénica que propusimos. El objetivo, de conocer la morfología del hueso normal de cráneo de ratones atímicos, fue desarrollar posteriormente una unidad osteogénica que reuniera las características de Osteogénesis, Osteoinducción y Osteoconducción, y, compararlas contra las que tiene dicho hueso normal durante su proceso de formación y remodelación. Así, realizamos un estudio piloto donde establecimos características morfológicas de hueso del cráneo de ratones atímicos, a través de tinciones histológicas específicas, con hematoxilina-eosina y von Kossa para buscar tejido osteoide y hueso mineralizado y Tricrómico de Massón para observar zonas osificadas. Además, analizamos el tejido óseo a través de inmunohistoquímica, con la finalidad de buscar marcadores de formación ósea como fosfatasa alcalina que es resultado del estímulo de la actividad osteoblástica; colágena 1, la cual es una proteína estructural de la matriz ósea; osteopontina, proteína sintetizada específicamente por osteoblastos que favorece la proliferación celular y la remodelación en defectos óseos; osteocalcina hormona peptídica producida por los osteoblastos durante la formación ósea y Runx 2 Factor de transcripción expresado por las células progenitoras que estimula la diferenciación ósea. Además, sometimos el tejido óseo a microscopía electrónica para determinar la presencia de depósitos de compuestos como calcio, fósforo y magnesio.

Estudio morfométrico del nervio óptico de tiburoncito (Ariopsis seemanni) / Morphological study of tete sea catfish (Ariopsis seemanni) optic nerve

En peces, el nervio óptico es el encargado de transportar la información integrada por las células ganglionares de la retina hacia el tectum óptico, para que se generen imágenes acerca del entorno. El objetivo de este trabajo es describir morfométricamente el nervio óptico del tiburoncito (Ariopsis seemanni), para lo cual se utilizó la Microscopía óptica de Alta Resolución (MOAR), realizando cortes a 1 micra de espesor. El nervio óptico de A. seemanni presenta fibras mielínicas de diverso calibre, acompañadas de oligodendrocitos y astrocitos. El nervio está cubierto por las meninges, que presentan vasos sanguíneos y adipocitos. El nervio tiene un área total de 179604 ± 30163 µm2, diámetro de 478 ± 42 µm y un número total de fibras mielínicas de 22848 ± 4350, de las cuales la mayoría tiene un tamaño pequeño, que puede estar relacionado con una velocidad de conducción baja.

The optic nerve carries out the information integrated by retinal ganglion cells towards the optic tectum, so that surrounding environment images are generated. The main goal of this paper is to describe morphometricaly the Tete sea catfish (Ariopsis seemanni) optic nerve, using high resolution optical microscopy (HROM), on 1 micron thick sections. A. seemanni optic nerve does present myelinated nerve fibers, accompanied by oligodendrocytes and astrocytes. The nerve is covered by the meninges, with blood vessels and adipocites. The nerve has a transversal area of 179,604 ± 30163 mm2, a diameter of 478 ± 42 mm and 22848 ± 4350 myelinated fibers, most are small in size, which may be related to a low conduction velocity.

Histological study on the role of ginger against cisplatin-induced testicular toxicity in albino rats

Chemotherapy with cisplatin has adverse effects on spermatogenesis. Therefore, this work aimed at investigating the protective role of ginger against cisplatin-induced testicular toxicity in male albino rats. Twenty-four adult albino rats were used in this study. They were divided into three groups. The first group served as the control group; the second group was injected with cisplatin [12 mg/kg once]; and the third group was injected with cisplatin [12 mg/kg once] and then given ginger [310 mg/kg orally] for 26 days. Testicular specimens were processed for light microscopic examination using H and E. Other specimens were processed for electron microscopic examination. Cisplatin had damaging effects on the seminiferous tubules. Some areas of the tubules showed complete depletion of germ cells. Other areas showed some spermatogonia or primary spermatocytes. Sertoli cells showed a variable degree of degenerative changes in the form of destruction of cellular processes and cell junction. Interruption of the nuclear envelope of spermatids and loss of intercellular bridges were noticed. Treating with ginger resulted in normal Sertoli cells and cell junctions. The germ cells lining the tubules were more or less normal except for some intercellular vacuolations. The use of ginger has some protective effects on the testicular structure; hence, a larger number of experiments with higher doses of ginger or longer administration period could be beneficial for patients taking chemotherapeutic drugs

Light and electron microscopic study on the effect of malathion on the adrenal gland of adult male albino rat and the possible protective role of vitamin [C]

Malathion is one of the organophosphorus insecticides widely used in agricultural and household applications to control pests. Actually, the studies of the effect of malathion on the adrenal%land are still limited, Vitamin C is the major water soluble antioxidant and free radical scavenger within the body. The aim of the work is to study the effect of chronic exposure to malathion on the adrenal gland. We also aim to study the possible protective effect of vitamin C in attenuating the nossible malathion induced changes. A total number of 30 adult male albino rats aged three months was used in the present study. Rats were randomly divided into three groups: l The first group [GI]: 10 rats were used as control 2- The second group [GH]: 10 animals were used as malathion treated group. 3- The third group [GUI]: 10 rats were used as malathion/vitamin C treated group. Commercial malathion was used, dissolved in distilled water and given orally by intragastric tube in a dose of 100 mg/kg/dayfor 2 months. Vitamin C was given orally by intragastric tube in a dose of 20 mg/WOgm/day for 2 months. After two months, the animals of the three groups were anaesthetized with ether inhalation and the suprarenal glands were dissected out and processed for light and electron microscopic examination. In malathion treated rats, there was cellulae disturbance in the arrangement of the adrenal gland. The cells of the cortex and medulla, showed irregular nuclei and apparent increase in the cytoplasmic vacuolation. Cortical and medullary blood capillaries were dilated and engorged with blood. Immunohistochemical staining for Caspase-3, showed many caspase-3 positive cells in the cortex and medulla. Ultrastructurally, degenerative changes were observed in the cortical and medullary cells in the form of cytoplasmic vacuolation, mitochondria! degeneration and increased lipid droplets. These changes were partially resolved by coadministration of vitamin C. Malathion had a harmful effect on adrenal gland so, it leads to impairment of its function in producing various hormones. This effect could be partially resolved, by concomitant administration of vitamin [C]. So, it is advisable to give vitamin [C] to those exposed to malathion

Modificaciones del endotelio corneal en cirugía de catarata simultánea con vitrectomía / Changes of the corneal endothelium in cataract surgery combined with vitrectomy

Métodos: se realizó un estudio observacional, descriptivo y prospectivo. El universo lo constituyó 15 pacientes (15 ojos) sometidos a cirugías de catarata y vítreo de forma simultánea. El estudio se efectúo en el Instituto Cubano de Oftalmología Ramón Pando Ferrer de enero a noviembre de 2010...

Methods: a prospective, descriptive and observational research was carried out in Ramón Pando Ferrer Cuban Institute of Ophthalmology from January to November 2010. The sample was made up of 15 eyes from 15 patients, who had undergone cataract surgery and vitrectomy simultaneously...

Diarreia aguda / Acute diarrhea

Diarreia aguda é a passagem de quantidade acima do normal de fezes amolecidas associada ao aumento do número de evacuações. No diagnóstico diferencial das diarreias agudas devem ser enfocados as infecções, as alergias alimentares, a intoxicação alimentar, o uso de medicações e a apresentação inicial de diarreia crônica. Dentre estas possíveis etiologias, especialmente em nosso meio, as causas infecciosas devem sempre vir à mente e constituir uma das primeiras opções na investigação diagnóstica. As infecções intestinais associadas a quadros diarreicos são a segunda causa de mortes de origem infecciosa em todo o mundo, com prevalência estimada de 3 a 5 bilhões de casos/ano. Os autores atualizam as novidades e peculiaridades a respeito do diagnóstico e dos tratamentos - geral e/ou específico - dos diferentes agentes associados à diarreia aguda infecciosa

Acute diarrhea is the passage of above normal quantities of soft faeces also associated with increased bowel movements. Differential diagnosis of acute diarrhea should be focused on infections, food allergies, food poisoning, use of medications and the initial presentation of chronic diarrhea. Among these possible etiologies, given the environment we live in, infectious causes should always be taken into account and be one of the first options in dignostic investigation. Intestinal infections associated with diarrheal frames are the second leading cause of infectious deaths worldwide, with an estimated to 3-5 billion cases/per year. In this review, the authors inted to review the new features and aspects concerning diagnosis and treatment - general and/or specific - of the different agents associated with acute infectious diarrhea

Age-related changes in the myenteric plexus of the gastric fundus in albino rats: histological and immunohistochemical study

Aging is believed to affect the structure and function of the enteric nervous system in the gastrointestinal tract. This work was designed to study the histological changes that might occur in the myenteric plexus of rat gastric fundus during aging. Thirty male albino rats were used in this study and divided equally into three groups: group A [which included 4-month-old rats], group B [which included 18-monthold rats], and group C [which included 24-month-old rats]. All the animals were anesthetized, and their stomachs were dissected out and processed for light and electron microscope examination. Also, immunohistochemical staining was carried out for the detection of protein gene product [PGP 9.5] and Bcl-2 proteins. The area% of myenteric ganglia was also determined. The results were analyzed statistically. With age, in group B [early senile], there was obvious neuronal loss and few gaps in the myenteric plexus, with a significant decrease in the number of PGP-positive neurons and moderate immunoreactions for the Bcl-2 protein. Glial cells had cytoplasmic vacuoles and nerve cells had peripheral condensation of heterochromatin in their nuclei. The myenteric plexus of the rats of group C [late senile] showed nerve cell bodies with apoptotic nuclei and cytoplasmic vacuolations. Many large cavities and eosinophillic cellular infiltration were observed within the neuropil. There was a marked apparent decrease in PGP 9.5-positive neurons and minimal immunoreactions for the Bcl-2 protein. Also, the area of myenteric ganglia increased with advancing age. In the current study, it was found that the myenteric plexus of the gastric fundus showed histological and immunohistochemical changes during aging, which may potentially be an explanation for the gastrointestinal tract disturbance in elderly individuals

Pregnancy-associated structural changes of the fallopian tube of rabbits with special reference to interstitial Cajal-like cells

The histological structure of the fallopian tube shows remarkable changes during pregnancy. Interstitial Cajal cell is known to play a key role in pacemaking motility in the gastrointestinal tract and the urinary tract. Its role in the motility of the fallopian tube is not fully explained. To describe the histological changes in the fallopian tube during early and late pregnancy, to shed light on the morphology of interstitial Cajal-like cells [ICLCs], and to localize their regional distribution in the fallopian tube in nonpregnant and pregnant rabbits. Thirty adult female rabbits were divided into three groups: group I [nonpregnant], group II [early pregnancy - day 4], and group III [late pregnancy - day 27]. The ampulla of the fallopian tube was excised and processed for histological, immunohistochemical, transmission, and scanning electron microscopy. In the nonpregnant group, it was found that the lining epithelium of the fallopian tube was composed of both ciliated and secretory cells. The ciliated cells were the major epithelial cell types; CD34-positive ICLCs were found maximally in the superficial zone of lamina propria, followed by musculosa, the deep portion of lamina propria, and least in serosa. The early pregnancy group showed a significant reduction in ciliated cells and an increase in secretory cells. In addition, there was a significant increase in the height of secretory cells and development of secretory organelles. There was a reduction in the number of CD34-positive ICLC in the superficial lamina propria compared with nonpregnant animals. The late pregnancy group showed an insignificant increase in ciliated cells and a decrease in secretory cells compared with group II. In addition, there was a decrease in the height of secretory cells and a reduction in their secretory organelles, but still more than that in group I. The structure of the fallopian tube showed histological changes during pregnancy. These changes represented a functional conditioning to the pregnancy state. ICLCs in the fallopian tube were prone to the same changes in their count and distribution

possible toxic effect of different doses of Nigella sativa oil on the histological structure of the liver and renal cortex of adult male albino rats

Nigella sativa seeds are commonly known as black seed or black cumin. It has been used for thousands of years as a spice and food preservative and also as a protective and curative remedy for numerous disorders. Our research aimed to study the possible toxic effect of different doses of N. sativa oil on the liver and kidney. Twenty-one adult male albino rats were used and divided equally into three groups. The first group was the control group. The second and third groups received the oil in two gradually increasing doses of 15 and 25 ml/kg, respectively, for 1 month. The animals were then sacrificed and samples of liver and kidney were taken and prepared for histological examination. In the kidney of group II there was epithelial shedding and necrosis of some cells of the proximal and distal convoluted tubules, but there was no effect on renal glomeruli. In contrast, in group III there was glomerular injury in the form of degeneration of the tuft of capillaries, ill-defined basement membrane, and destruction of endothelial cells, in addition to tubular necrosis. In group II there was minimal effect on the liver in the form of perivascular cellular infiltration; in group III was seen a markedly vacuolated foamy cytoplasm of hepatocytes, with dilated sinusoids and perivascular cellular infiltration. In conclusion, large doses of N. sativa oil have toxic effects on the histological structure of the kidney and to a lesser degree on the liver. Therefore, Nigella oil should be used in proper doses, and further studies on the effect of large doses of oil are recommended

Effect of green tea on aged rat skeletal muscle: a light and electron microscopic study

Age-related sarcopenia represents a powerful risk factor for the loss of independence and physical disability in the elderly. Green tea contains a series of polyphenols called catechins and has been applied for disease prevention and treatment. To determine the effects of the intake of green tea on age-related changes in skeletal muscle. Thirty female albino rats were divided into three equal groups: group I [control adults] included 6-month-old rats, group II [aged]included 18-month-old rats, and group III [green tea treated] included 12-month-old rats that received green tea at a dose of 300mg/kg body weight in 1.5ml distilled water orally daily by a gastric tube for 6 months. Quadriceps muscle was dissected bilaterally and prepared for light and electron microscopical examination. Morphometric and statistical studies of the mean cross-sectional area of myofibers were carried out in the three studied groups. With increasing age, there were focal degeneration and atrophy, and significant decrease in the mean cross-sectional area of myofibers. The connective tissue framework showed increased collagen fiber deposition and deficient reticular fibers. Ultrathin sections showed areas of myofibrillar loss and mitochondria with destroyed cristae. Dense shrunken nuclei of satellite cells were observed. After treatment with green tea, normal-appearing myofibers, except for centrally located nuclei, were observed. The mean cross-sectional area of myofibers increased significantly compared with the aged group. Collagen and reticular fibers were relatively similar to those of the control. Proliferation of satellite cells was observed with restoration of normal myofiber ultrastructure. Good regenerative outcome was observed in aged skeletal muscle after the intake of green tea

Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A

Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi. In the following investigation, a novel chitinase with antifungal activity was characterized from a native Serratia marcescens B4A. Partially purified enzyme had an apparent molecular mass of 54 kDa. It indicated an optimum activity in pH 5 at 45ºC. Enzyme was stable in 55ºC for 20 min and at a pH range of 3-9 for 90 min at 25ºC. When the temperature was raised to 60ºC, it might affect the structure of enzymes lead to reduction of chitinase activity. Moreover, the Km and Vmax values for chitin were 8.3 mg/ml and 2.4 mmol/min, respectively. Additionally, the effect of some cations and chemical compounds were found to stimulate the chitinase activity. In addition, Iodoacetamide and Idoacetic acid did not inhibit enzyme activity, indicating that cysteine residues are not part of the catalytic site of chitinase. Finally, chitinase activity was further monitored by scanning electronic microscopy data in which progressive changes in chitin porosity appeared upon treatment with chitinase. This enzyme exhibited antifungal activity against Rhizoctonia solani, Bipolaris sp, Alternaria raphani, Alternaria brassicicola, revealing a potential application for the industry with potentially exploitable significance. Fungal chitin shows some special features, in particular with respect to chemical structure. Difference in chitinolytic ability must result from the subsite structure in the enzyme binding cleft. This implies that why the enzyme didn't have significant antifungal activity against other Fungi.