Anatomy and histology of the digestive tract and immunolocalization of Npy in the fish Cichlasoma dimerus (Cichliformes: Cichlidae) / Anatomía e histología del tracto digestivo e inmunolocalización del Npy (péptido orexigénico) en el pez Cichlasoma dimerus (Cichliformes: Cichlidae)

Abstract Introduction: The digestive tract of Neotropical cichlids has not been deeply studied, and it is a fundamental topic for understanding fish physiology, nutrition, trophic associations, and evolution. Objective: To describe anatomically and histologically the digestive tract of the Neotropical cichlid fish Cichlasoma dimerus and to immunolocalize the orexigenic peptide (Npy) along the intestine. Methods: We euthanized 14 adult individuals and fixed the organs in Bouin´s solution; we stained 7 μm thick paraffin sections for general description and with Alcian Blue (pH = 2.5, AB) and Periodic acid-Schiff (PAS) to identify acid or neutral glycoconjugates, respectively. Additionally, we performed immunohistochemistry for Npy in 3 adult individuals. We manually counted PAS- and AB-positive cells, and Npy-immunoreactive cells per fold. Results: There is a short oesophagus, a sac-like stomach, and a tubular intestine with two loops. The oesophagus has a stratified epithelium with a high density of PAS- and AB-positive goblet cells and striated muscle fibers in the tunica muscularis. The stomach mucosa is formed by simple columnar epithelium. The intestine has a simple columnar epithelium, with brush border and interspersed PAS- and AB-positive goblet cells, and Npy-immunoreactive cells. There is an ileorectal valve in the transition between the posterior intestine and the rectum. This last gut portion has goblet cells and a thicker tunica muscularis. Conclusions: C. dimerus shares features with other Neotropical cichlids, but the goblet cells and gastric glands distribution seems to be unique for the species. To our understanding, this is the first work to describe Npy-immunoreactive cells distribution in the intestine of a Neotropical cichlid fish.

Resumen Introducción: El tracto digestivo de los cíclidos neotropicales no ha sido profundamente estudiado y es un tema fundamental para entender la fisiología, nutrición, asociaciones tróficas y evolución de los peces. Objetivo: Describir anatómica e histológicamente el tracto digestivo del pez cíclido neotropical Cichlasoma dimerus e inmunolocalizar el péptido orexigénico (Npy) a lo largo del intestino. Métodos: Sacrificamos 14 individuos adultos y fijamos los órganos en solución de Bouin; teñimos secciones de parafina de 7 μm de espesor para una descripción general y con azul alcián (pH = 2.5, AB) y ácido periódico-Schiff (PAS) para identificar glicoconjugados ácidos o neutros, respectivamente. Además, en 3 individuos adultos se realizaron inmunohistoquímicas contra Npy. Contamos manualmente las células PAS y AB positivas, y las células inmunorreactivas a Npy por pliegue. Resultados: Hay un esófago corto, un estómago en forma de saco y un intestino con dos vueltas. El esófago tiene un epitelio estratificado con una alta densidad de células caliciformes PAS- y AB- positivas y fibras esqueléticas estriadas en las capas musculares. La mucosa del estómago está revestida por epitelio simple cilíndrico. El epitelio intestinal es simple cilíndrico con chapa estriada y células caliciformes PAS- y AB- positivas intercaladas, y células inmunorreactivas a Npy. Hay una válvula ileorrectal en la transición entre el intestino posterior y el recto. Esta última porción intestinal tiene células caliciformes y una túnica muscular más gruesa. Conclusiones: C. dimerus comparte características con otros cíclidos neotropicales, pero la distribución de las células caliciformes y las glándulas gástricas, serían rasgos propios de esta especie. A nuestro entender, este es el primer trabajo que describe la distribución de células inmunorreactivas a Npy en el intestino de un pez cíclido neotropical.

Congenital myopathies: The current status

Within the history of neuromuscular diseases (NMD), congenital myopathies (CM) represent a relatively new category introduced in the mid-nineteen hundreds upon advent and subsequent application of enzyme histochemistry and electron microscopy by establishing the three major CM, central core disease, nemaline myopathy, and centronuclear myopathy which later pluralized each when the molecular era began at the end of last century. Quickly, during the following 5 decades, many new CM entities were described, based on muscle biopsies and their CM-characteristic myopathology, the former a prerequisite to recognizing an individual CM, the latter of the nosological hallmark of the individual CM. When the molecular era ushered in immunohistochemistry the spectrum and nosography of CM altered in that some CM became allelic to other cohorts of NMD, e.g., congenital muscular dystrophies, other muscular dystrophies, distal myopathies based on different or identical mutations in the same gene. The nosological spectrum of a defective gene also enlarged by recognizing several entities with mutations in the same gene, and same or similar nosological conditions originated from mutations in different genes. Lately, however, CM were reported which lacked any individual myopathological hallmarks, but were clearly based on molecular defects, a fair number of them being newly identified ones. Few CM still remain without any molecular clarification. This nosographic development rendered the original definition of such new CM questionable and brought uncertainty to their classification and nomenclature.

Basic requirements to establish a neuromuscular laboratory

Histopathological analysis of muscle biopsy is a prerequisite in the evaluation of neuromuscular disorders, particularly inflammatory myopathies, metabolic myopathies, congenital myopathies, muscular dystrophies and differentiating myopathies and neurogenic disorders with overlapping clinically features. It not only provides useful information that helps in the diagnosis but also treatment and management. Fundamental skills and basic knowledge regarding handling, processing and analyzing a muscle biopsy are required in any specialized or a general pathology lab supporting neuromuscular clinical services. Care during transport of the muscle biopsy, sample receipt in the laboratory and grossing is very important. Standard operating procedure should be followed for the preanalytical steps (freezing and cryomicrotomy), routine and special staining (enzyme and non enzymatic) and immunohistochemistry. A well organized neuromuscular laboratory with good quality management system is necessary for the practice of myopathology. This article gives an overview of establishing such a laboratory.

Micromorfología y ultraestructura de las anteras y los granos de polen en diez genotipos élite de Theobroma cacao (Malvaceae) / Micromorphology and ultrastructure of anthers and pollen grains in ten elite genotypes of Theobroma cacao (Malvaceae)

Resumen Introducción: A pesar de que T. cacao es una especie importante a nivel mundial por la producción de cacao, es poco lo que se conoce sobre la micromorfología y estructura de las anteras y los granos de polen. Objetivos: Describir y analizar la estructura y micromorfología de las anteras y los granos de polen de 10 genotipos élite de esta importante especie tropical. Métodos: Se tomaron más de 30 anteras de flores en antesis de los 10 genotipos élite de T. cacao del banco de germoplasma ex situ del Centro de Investigaciones Suiza-Agrosavia (Rionegro, Santander-Colombia). El material se procesó de acuerdo con los protocolos estándar para embeber y seccionar en parafina. Las secciones obtenidas (3 μm) se tiñeron con azul de Safranina-Alcian para discriminar estructuras con paredes primarias y secundarias y polifenoles totales. Además, se usó la técnica PAS-Amidoblack para diferenciar entre polisacáridos estructurales y de reserva, así como proteínas. Para la determinación de esporopolenina y polifenoles se usó la tinción azul de toluidina y finalmente para descripciones adicionales se aplicó la tinción azul alcián-PAS-hematoxilina. Las observaciones se realizaron mediante microscopio fotónico y microscopio de epifluorescencia. Para la observación con microscopía electrónica de barrido (MEB), las anteras con los granos de polen se fijaron y deshidrataron en 2.2 dimetoxipropano, luego se desecaron hasta un punto crítico y finalmente se recubrieron con oro. Resultados: Las anteras son bitecas y están sostenidas por un largo filamento formado por un estrato epidérmico, tejido parenquimatoso y un haz vascular. La dehiscencia ocurre longitudinalmente a través del estomio. La pared de la antera madura está formada por una capa epidérmica monoestratificada, una capa de células endoteliales con engrosamientos fibrilares lignificados y se pueden apreciar restos celulares del tapete y abundantes orbículas recubriendo la cavidad de los microesporangios. Los tejidos epidérmicos y parenquimatosos de las anteras almacenan polifenoles. Las orbículas son generalmente esféricas, psiladas y exhiben las mismas reacciones de tinción y fluorescencia que la exina de los granos de polen. Los granos de polen son mónades, isopolares, pequeños (16-19 µm) con amb circular, esferoidales, tricolpados con colpos medianos o cortos (5-10 µm) con membrana ornamentada, semitectatos, reticulados, heterobrochados, las paredes del retículo ornamentadas o no, con microgránulos de diferente tamaño o escabrados. Los análisis estadísticos mostraron que existen diferencias significativas en el tamaño de los granos de polen (P ˂ 0.05). Se observa que los granos de polen más pequeños son los del genotipo TCS 19 (16.890 µm) y se diferencian del resto de genotipos, y entre estos no se observan diferencias significativas. Solo dos genotipos (SCC 19 y SCA 6) presentaron polenkit y solo uno tiene paredes perforadas (SCA 6). Conclusiones: La estructura y micromorfología de las anteras de T. cacao son similares a las descritas para otras Malvaceae. Así mismo, los granos de polen mostraron variaciones de tamaño, ornamentación de las paredes y del lumen del retículo y presencia de polenkit. Sin embargo, no se observó relación entre las variaciones de los caracteres micromorfológicos analizados en los granos de polen y los modelos de compatibilidad polínica reportados para estos genotipos.

Abstract Introduction: Despite the fact that T. cacao is an important species worldwide for cocoa production, little is known about the micromorphology and structure of anthers and pollen grains. Objectives: To describe and analyze the structure and micromorphology of the anthers and pollen grains of 10 elite genotypes of this important tropical species. Methods: More than 30 anthers of flowers in anthesis were taken of the 10 elite genotypes of T. cacao from the ex situ germplasm bank of the Suiza-Agrosavia Research Center (Rionegro, Santander-Colombia). The anthers with the pollen grains were fixated and processed according to the standard protocols for embedding and sectioning in paraffin. Sections obtained (3 μm thick) were stained with Safranin-Alcian blue to discriminate structures with primary and secondary walls and total polyphenols. Additionally, the samples were also stained with the PAS-Amidoblack technique was used to differentiate between structural and reserve polysaccharides as well as proteins. Toluidine blue staining was used for the determination of sporopollenin and polyphenols and finally Alcian blue-PAS-Hematoxylin staining was applied for additional descriptions. Observations were made using photonic microscopy and epifluorescence microscopy. For observation with scanning electron microscopy (SEM) the anthers with the pollen grains were fixed and dehydrated in 2.2 Dimethoxypropane, then desiccated to critical point and finally coated with gold. Results: The anthers are dithecal and supported by a long filament made up of an epidermal stratum, parenchymal tissue, and a vascular bundle. The dehiscence occurs longitudinally through the stomium. The anther wall is made up of a monostratified epidermal layer, followed by a layer of endothecial cells with lignified fibrillar thickenings, cellular remnants of tapetum and abundant orbicules can be seen covering the cavity of the microsporangia. The epidermal and parenchymal tissues of the anthers are abundant in polyphenols. Orbicules are generally spherical, psilated, and these exhibit the same staining and fluorescence reactions as exine from pollen grains. The pollen grains are monades, isopolar, small (16-19 µm) with circular amb, spheroidal, tricolpate with medium or short colpi (5-10 µm) with sculptured membrane, semitectate, reticulated, heterobrochate, sculptured or non- sculptured walls, with microgranules of different size or scabrate. The statistical analyzes showed that there are significant differences in the size of the pollen grains (P ˂ 0.05). It is observed that the smallest pollen grains are those of the TCS 19 genotype (16.890 µm) and are different from the other genotypes, and among these there are no significant differences. Only two genotypes (SCC 19 and SCA 6) showed pollenkit and only one has perforated walls (SCA 6). Conclusions: The structure and micromorphology of the anthers of T. cacao are similar to those described for other Malvaceae. Likewise, the pollen grains showed variations in size, ornamentation of the sporoderm and the lumen of the reticulum and the presence of pollenkitt. However, no relationship was observed between the micromorphological characters analyzed in the pollen grains and the pollen compatibility models reported for these genotypes.

Anatomical and histochemical descriptions of the male reproductive system of Diachasmimorpha longicaudata (Ashmed) (Hymenoptera: Braconidae)

ABSTRACT The parasitoid Diachasmimorpha longicaudata is an important control agent for several species of fruit flies. Research on the characteristics of the reproductive system and reproductive biology of this parasitoid can be valuable for studies in taxonomy and phylogeny of insects of the order Hymenoptera. In this study we analyzed the histology and histochemistry of the male reproductive system of D. longicaudata. In this species the male reproductive system consists of a pair of testes, two accessory glands, and an ejaculatory duct. Each testicle consists of only one follicle. The testicular follicles are filled with cysts in different stages of spermatogenesis. Histochemical analyses detected proteins and carbohydrates in the cytoplasm of secretory cells and in the lumen of accessory glands. The morphology of the male reproductive system of D. longicaudata differs in some respects from other species of Hymenoptera.

Eucalyptus cinerea: Microscopic Profile, Chemical Composition of Essential Oil and its Antioxidant, Microbiological and Cytotoxic Activities

Abstract Eucalyptus species possess anti-inflammatory, antifungal, antibacterial, and insecticidal properties. In this study, the chemical composition and biological activities of Eucalyptus cinerea essential oil (EO) and the leaf and stem anatomy were investigated. EO was extracted by Clevenger apparatus and the compounds were identified by GC/MS. The antioxidant activity was evaluated by DPPH, ABTS, and reducing phosphomolybdenum complex. Broth microdilution was used to determine antimicrobial activity. Cytotoxicity was verified against HeLa, HRT-18, and Calu-3 cells by MTT assay. The cytotoxic mechanism was studied by cell DNA content, cell cycle, and DNA fragmentation. The microscopic analyzes of the leaves and the stems were performed by light microscopy, field emission scanning electron microscopy, and energy-dispersive X-ray spectroscopy. The main constituent of the EO was 1,8-cineole (55.24%). The EO showed low antioxidant and antimicrobial activities. Calu-3 cells showed a significant reduction in viability with IC50 of 689.79 ± 29.34 μg/mL. EO at 1000 μg/mL decreased the DNA content in Jurkat cells. In general, EO increased cell percentage in sub-G0 and S phases with concomitant reduction of cell percentage in G0/G1 and G2/M phases and provided DNA fragmentation of 29.73%. Anatomical and micromorphological features of the leaves and stems can help in the species identification and its differentiation from other Eucalyptus species.

A histological and histochemical study on the gallbladder of the Alburnus tarichi (Güldenstädt, 1814) (Cyprinidae) / Estudio histológico e histoquímico de la vesícula biliar de Alburnus tarichi (Güldenstädt, 1814) (Cyprinidae)

In the current study, the histological structure of the gallbladder of Alburnus tarichi (Güldenstädt, 1814) was investigated. Hematoxylin and eosin were used to stain the histological sections for routine examinations, in addition to using periodic acid Schiff (PAS) for the neutral mucins, aldehyde fuchsin (AF) for the sulphated mucins, and Alcian blue (AB; pH: 2.5) for the acidic mucins. In addition, proliferating cell nuclear antigen (PCNA) immune-staining was performed for the detection of dividing cells among the epithelium. The gallbladder of A. tarichi was composed of mucosa, muscularis, and serosa or adventitia layers. The mucosa covering the wavy pleomorphic folds was made up of tall columnar epithelium and a lamina propria. The apical surface of the epithelial cells was lined by continuous short microvilli. On the epithelium, the luminal surface was remarkably stained with PAS, AF, and AB. Slight to moderate staining was observed on the epithelial cells in the apical zone with PAS. The cytoplasm of epithelial cells were stained in a slight manner with AF. No goblet cells were observed among the epithelium. According to the PCNA immune-staining, some epithelial cells were observed to proliferate. The lamina propria consisted of loose connective tissue that contained fibrocytes, collagen and elastic fibers, capillaries, and small blood vessels. The muscularis layer displayed muscle fibers that were circular, smooth, and surrounded by collagen fibers. The subserosal and serosal or adventitial layers had typical morphology to those of other fish and vertebrates.

En este estudio, se investigó la estructura histológica de la vesícula biliar de Alburnus tarichi (Güldenstädt, 1814). Las secciones histológicas se tiñeron con Hematoxilina-Eosina para los exámenes de rutina, además de usar el ácido periódico de Schiff (PAS) para las mucinas neutras, aldehído fucsina (FA) para las mucinas sulfatadas y azul alcián (AB; pH: 2,5) para las mucinas ácidas. Además, se realizó una tinción inmune de antígeno nuclear de células proliferativas (PCNA) para la detección de células en división entre el epitelio. La vesícula biliar de A. tarichi estaba compuesta de capas, mucosa, muscular y serosa o adventicia. La mucosa que cubría los pliegues pleomórficos ondulados estaba formada por un epitelio columnar alto y una lámina propia. Se observó una superficie apical de las células epiteliales revestida por microvellosidades cortas y continuas. En el epitelio se observó una tinción importante de la superficie luminal teñida con PAS, FA y AB. Se observó una tinción leve a moderada en las células epiteliales en la zona apical con PAS. El citoplasma de las células epiteliales se tiñó ligeramente con FA. No se observaron células caliciformes entre el epitelio. Según la tinción de PCNA, se observó que proliferaban algunas células epiteliales. La lámina propia consistía en tejido conectivo laxo que contenía fibrocitos, colágeno y fibras elásticas, capilares y pequeños vasos sanguíneos. La capa muscular mostraba fibras musculares circulares, lisas y rodeadas de fibras de colágeno. Las capas subserosas y serosas o adventicias tenían una morfología típica a la de otros peces y vertebrados.

Sarcoma fibromixoide intracraneal primario de bajo grado: reporte de caso y revisión de la literatura / Low-grade fibro-myxoid sarcoma of intracranial origin: A case report and literature review

Resumen El sarcoma fibromixoide de bajo grado (LGFMS) es un tumor de tejidos blandos de origen mesenquimal. Los sarcomas son un grupo heterogéneo, que representa el 1% de todas las neoplasias. Los sarcomas primarios del sistema nervioso central (SNC) son raros y representan solo el 0,7% del total de sarcomas, con una incidencia estimada de 3 por cada 10 millones de personas por año. En este artículo, se describe el caso de una mujer de 59 años que presentó un sarcoma fibromixoide intracraneal de bajo grado, localizado en la región parietal derecha. Se discute el curso clínico, estudios de imágenes, características histopatológicas y tratamiento de este diagnóstico infrecuente y, por lo mismo, muy poco reportado. El diagnóstico definitivo se obtiene, ciertamente, mediante estudios histo-patológicos.

Summary Low-grade fibromyxoid sarcoma (LGFMS) is a soft tissue tumor of mesenchymal origin. Sarcomas are a heterogeneous group, representing 1% of all neoplasm diagnoses. Primary sarcomas of the central nervous system (CNS) are rare, and represent only 0.7% of all sarcomas, with an estimated incidence of 3 per 10 million people per year. The case of a 59-year-old woman who developed a low-grade intracranial fibromyxoid sarcoma in the right parietal region, is described. The clinical course, imaging studies, histopathological features, and treatment approach of this unusual diagnosis, are discussed. Low-grade intracranial fibromyxoid sarcoma is a rare and probably under- reported condition. The definitive diagnosis is usually made through histo-pathological studies.

Desmoplastic small round cell tumor of the ovary: A case report with a new modality of treatment and review of the literature

Abstract Objective Desmoplastic small round cell tumor (DSRCT) is a rare intraabdominal neoplasm that grows along serosal surfaces and is primarily found in young men. To Keywords date, only 16 cases of ovarian DSRCT have been previously reported in women in the English literature, and no large population-based studies on this topic exist. Case Report We report the case of a 19-year-old virgo with unremarkable past medical history, initially presented with abdominal fullness. After being treated with the optimal treatment modality (primary and secondary surgical debulking, unique chemotherapy, protocol and adjuvant radiotherapy), the patient has remained without tumor disease for 40 months. Conclusion Although the best therapy for patients with DSRCT has yet to be determined, combining complete surgical resection, adjuvant chemotherapy, and radiotherapy is required to prolong survival and to achieve proper quality of life.

Role of Ki 67 Labelling Index as an Adjunct to Histopathological Diagnosis for Grading of CNS Tumours

The Central Nervous System tumours are unique as they arise from specialized tissue. CNS tumours constitute a wide range of neoplasm that differs in their location, age distribution, extent of invasiveness, morphological features and tendency for progression. We wanted to evaluate the traditional morphological data with knowledge on the prognostication marker Ki 67 antibody in evaluating tumour grade and prognosis of CNS neoplasm.METHODSThis is a cross section study carried out between March 2015 and September 2016 in histopathology department of Dhiraj Hospital on 50 cases of CNS tumours. Immunolabelling of all biopsies was done by horse radish peroxidase technique using rabbit monoclonal antibody to Ki 67 (clone SP 6) (Thermo Scientific, USA). Ki 67 immune positive labelling index was obtained for each tumour and correlated with mitotic labelling index obtained by conventional morphological grading as per WHO 2007 classification.RESULTSIn our study of CNS tumours, all age groups were studied. The mean Ki 67 labelling index (LI) values +/-SD for WHO Grade I tumours were- meningiomas (10) 3.85 (+/1.97) %, schwannoma (3) -3.0 (+/-2.97) %, pituitary adenoma (1) 0.6, craniopharyngioma (1) -1.1% and ependymoma (6) 2.62 (+/-0.60) %. WHO Grade II tumours- atypical angiomatous meningioma (1) -2%, atypical mucinous meningioma (2) -6.15 (+/-1.06) %, gemistocytic astrocytoma (1) -12; pleomorphic xanthoastrocytoma (2) -4.3 (+/-0.99) %, astrocytoma grade ii (2) -3.3 (+/-0.71) %, oligodendroglioma grade ii (4) - 3.9 (+/-0.88) %. WHO grade III tumours- anaplastic astrocytomas (5) -6.82 (+/-2.17) %, anaplastic oligoastrocytoma grade iii (1) -10.8 %. who grade iv-glioblastomas (7) -18.44 (+/-3.97) %; medulloblastomas (1) 20%, metastatic tumour (3) -36 (+/- 22.16) %. In our study, the mean Ki 67 LI (± SD) values for grade II, III and IV glioma is as follows: 4.76 (+/-2.83) %, 7.48 (+/-2.53) % and 18.44 (3.97) % respectively.CONCLUSIONSThis study shows that Ki 67 LI serves as an essential clinical prognostic proliferation marker of particular importance in cases with lower grade histology of Grade II & Grade III astrocytomas, Grade II & Grade III oligodendrogliomas. Ki 67 LI is important in determining benign, atypical and malignant meningiomas, non-invasive and invasive pituitary adenomas.

Métodos histoquímicos e inmunohistoquímicos para la estimación del Intervalo postmortem en tejidos humanos: una revisión / Histochemical and immunohistochemical methods for the postmortem interval estimation in human tissues: a review

El intervalo postmortem (IPM) es un importante desafío a resolver en patología forense, y consiste en poder determinar el tiempo transcurrido desde la muerte hasta el momento de la autopsia. Dada la poca confiablidad de algunos métodos por la gran influencia de factores externos, la Histoquímica (HQ) y la Inmunohistoquímica (IHQ), entre otros, han recibido considerable atención por sus niveles de objetividad en la investigación forense. Se presenta una revisión con búsqueda sistemática de estudios experimentales que apliquen métodos HQs e IHQs para la estimación del IPM sobre material cadavérico humano. Se identificaron 1053 artículos de los cuales 12 cumplieron con los criterios, a los que se agregaron 4 mediante una búsqueda manual (n=16 artículos). Alemania fue el país con más publicaciones destacando con 8 artículos. Las técnicas HQs de AgNORs, TRAP y PAS fueron utilizadas en 5 estudios (glándulas sudoríparas, piel, hígado, médula ósea y mucosa labial), mientras que las IHQs fueron empleadas con diferentes grupos antigénicos en 12 estudios (páncreas, cerebro, pulmón, tiroides, hígado, glándulas pituitarias, glándulas sudoríparas y mucosa gingival). Las estimaciones del IPM fueron posibles con márgenes entre 2-3 h. hasta los 20 días dependiendo de la técnica. El análisis de tejidos provenientes de cavidad oral asegura una vía no invasiva, de fácil acceso y bajo resguardo natural a la influencia de factores ambientales. Si bien no existe un método único que permita de manera confiable estas estimaciones, la introducción de nuevas técnicas permitiría evitar la producción de errores.

The postmortem interval (IPM) is an important challenge to be solved in forensic pathology, and it consists in determine the time elapsed since death until the autopsy. Given the low reliability of some methods due to the great influence of external factors, Histochemistry (HQ) and Immunohistochemistry (IHQ), among others, have received considerable attention for their levels of objectivity in forensic investigation. A scoping review of experimental studies that apply HQs and IHQs methods to estimate the MPI on human cadaveric material is presented. We identified 1053 articles, of which 12 met the criteria; we added 4 articles through a manual search (n = 16 articles). Germany was the most productive country, with 8 articles. HQ techniques of AgNORs, TRAP and PAS were used in 5 studies (on sweat glands, skin, liver, bone marrow and labial mucosa), while IHQs techniques were used with different antigenic groups in 12 studies (on pancreas, brain, lung, thyroid, liver, pituitary glands, sweat glands and gingival mucosa). IPM estimates were possible with margins between 2-3 hours up to 20 days depending on the technique. The analysis of oral tissues ensures a non-invasive route, easily accessible and under natural protection to the influence of environmental factors. Although there is no single method that reliably allows these estimates, the introduction of new techniques would prevent the production of errors.

Hepatotoxic Effect of Port Harcourt Eliozu Landfill Leachate in Wistar Rats

Studies have reported deleterious effect of landfill leachate and their surrounding water sources around the world. None has been reported on the Port Harcourt (Eliozu) landfill. This therefore necessitates this study that evaluated the hepatotoxiceffects of Port Harcourt (Eliozu) landfill in wistar rats. 25 wistar rats were grouped into five of five rats each. Group 1 served as the control and received 1ml of commercial non-carbonated bottled water; Group 2 served as negative control and received 1ml of water obtained from borehole about 1km from the landfill; while Groups 3,4 and 5 received 1ml of 10% of leachate concentration, 1ml of 50% of leachate concentration and 1ml of 100% of leachate concentration respectivelyonce daily for90 days. After the treatment period, the rats were sacrificed and 5ml of blood was collected via cardiac puncture. The blood samples were prepared and used for the determination of serum concentrations of liver enzymes. The rats were also dissected and the liver harvested for histochemical analysis using the periodic acidSchiff stain. Result showedthe serum concentrations liver enzymes significantly increased in the rats exposed to the leachate and water from a nearby borehole. Also histochemistry examination showed a marked distortion of the hepatic cells and cirrhotic changein the test groups compared to control. This study therefore concludes that exposure to Port Harcourt Eliozu landfill leachate (PELL) may cause significant hepatotoxicity in rats

Histoarchitecture and Histochemical Studies on the Lacrimal Gland of Pig (Sus scrofa)

The fresh tissues were collected from five pigs of either sex and processed for histomorphological studies. The gland was located on dorso-lateral aspect of the eye in the orbital cavity. The gland was multilobar tubulo-acinar with predominance of acini. It was covered by a thick connective tissue capsule and connective tissue septa, which penetrate into the glandular parenchyma and divided the gland into lobes and lobules. The acini were of mixed type but predominantly sero-mucous in nature. The secretory units were followed by a duct system. The excretory duct system was made up of intercalated, intralobular, interlobular and large excretory ducts. The strong presence of glycogen in the acini was demonstrated with the help PAS (McManus’) stain. The ducts showed very weak reaction for the presence of glycogen but goblet cells in the stratified ducts showed strong positive reaction for PAS. The acini showed strong reaction for acidic mucosubstances with PAS-AB. The goblet cells in the stratified ducts also showed strong positive activity towards the PAS-AB. The intense activity towards the Alcian blue showed presence of sialomucins and hyaluronic acids in glandular parenchyma of the lacrimal gland

Extrarenal angiomyolipoma in uterine cervix: rare presentation in unusual site

Angiomyolipoma is a benign neoplasm composed of variable admixture of blood vessels, smooth muscle cells and adipose tissue. Cervical angiomyolipoma are extremely rare and to the best of our knowledge only five cases of angiomyolipoma in cervix have been reported in the literature till date. Authors are presenting a case of angiomyolipoma arising from the uterine cervix. 43 years old female presented with mass descending per vagina for 6 months. This case had no association with tuberous sclerosis. Microscopic examination showed an ill-defined polypoidal, non-encapsulated lesion covered by keratinized stratified epithelium. The lesion is made up of three components, predominantly by fascicles of spindle shaped cells, varying sized blood vessels and multiple foci of mature adipocytes with no evidence of atypia or increased mitotic activity. Smooth muscle component showed strong immunoreactivity to SMA and absence of elastic fibres in the blood vessels were confirmed by histochemistry. Non-vascular smooth muscle cells were negative for HMB-45 in contrast to renal and other extra-renal angiomyolipoma in which HMB-45 immunoreactivity in seen in these cells. To conclude, the differential diagnosis of lower abdominal mass and dysfunctional uterine bleeding should include the angiomyolipoma, even though the uterine cervix is an extremely rare location where they occur.

Anatomía general, histología y morfometría del sistema digestivo del pez Pterophyllum scalare (Perciformes: Cichlidae)

Introducción: El pez escalar (Pterophyllum scalare) es una de las principales especies de peces ornamentales comercializados de Sur América. Objetivo: Describir histológica y morfométricamente el tracto digestivo del pez tropical P. scalare. Métodos: Se utilizaron 19 individuos adultos, y se calculó el coeficiente intestinal. Se realizó una descripción macroscópica e histológica del esófago, estómago e intestino (región anterior parte a, anterior parte b, media y posterior). Las muestras fueron procesadas en parafina, se obtuvieron cortes de 5 µm, que fueron teñidos con H&E y P.A.S. Se realizó conteo de células P.A.S +. Para la morfometría se digitalizaron los cortes y se midió el área y perímetro del lumen, y el área tisular para cada región. Resultados: P. scalare presenta un esófago corto, un estómago en forma de saco ciego poco muscular junto a un intestino con una longitud similar a su tamaño, coeficiente intestinal (1.66 +/- 0.52). Se identificaron cuatro capas constitutivas: mucosa, submucosa, muscular y en algunas regiones serosa. El esófago se destacó por ser la región con mayor número de células caliciformes P.A.S. + y mayor área tisular. En cuanto al área y perímetro luminar, la región anterior parte a del intestino fue la de mayor valor. Conclusiones: A partir del coeficiente intestinal (1.66 +/- 0.52), y las características histológicas y morfométricas de los órganos evaluados, se puede decir que P. scalare presenta un hábito alimenticio omnívoro con preferencia de presas blandas.

Introduction: Ornamental Angel fish (Pterophyllum scalare) is one of the main commercialized species from South America, with an important number of national and international dealers. Objective: To make a histological and morphometric description of P. scalare digestive tract. Methods: Nineteen adult individuals were used. Intestinal coefficient values were obtained, and a macroscopic and microscopic description of esophagus, stomach, and intestine (anterior, middle and posterior) was carried out. Samples were processed for paraffin embedding, and 5 µm sections were stained with H&E and P.A.S. Positive P.A.S. cells were counted. Lumen area and tissue area were measured in representative digitalized sections from each region. Results: P. scalare exhibited a short esophagus, followed by a blind bag shaped, scarcely muscular stomach, and a short intestine (intestinal coefficient 1.66 +/- 0.52). Four constitutive tissue layers were identified: mucosa, submucosa, muscular and in some areas also a serosa layer. Esophagus exhibit abundant globet P.A.S. + cells and shows the larger tisular area was observed. Luminal area and perimeter are larger in anterior intestinal region. Conclusions: The intestinal coefficient value (1.66 +/- 0.52), and the histological and morphometric characteristics of the evaluated organs, suggest that P. scalare is omnivorous with soft prey preferences.

Microscopic histochemical comparison during mountain-agarwood formation / 中国中药杂志

Mountain-agarwood plays an important role in ethnic medicine in China for its pharmaceutical value. Modern pharmacological researches demonstrated that mountain-agarwood was effective for its anti-myocardial ischemia, antibacterial, anti-inflammatory, antitumor and analgesic effects. Mountain-agarwood derives from the peeled roots, stems or twigs of Syringa pinnatifolia which belongs to Syringa genus. It often depends on the purple substance and fragrance to estimate the formation of mountain-agarwood. However, the mechanism of mountain-agarwood formation has not been reported. To observe the microcosmic change in the process during the formation of mountain-agarwood, this study described the microscopic and histochemical characteristics of mountain-agarwood formation through histochemical staining. Our results showed that a significant difference of the distribution of tyloses existed during mountain-agarwood formation. It was observed that inchoate mountain-agarwood had more starch granules and viable cells than mountain-agarwood formed with high level or low level. The amount of polysaccharide and degree of lignification were increased during the mountain-agarwood formation. The results indicated that the mountain-agarwood, which meets the quality requirements for pharmaceutical use, contained the following characteristics: a large amount of purple tyloses in heartwood; yellow-brown tyloses distributing in heartwood and sapwood which were less in the latter; lignification with high level; a few viable cells; lots of polysaccharide and few starch granules in xylem rays cell. This study is aimed to reveal the change of histochemical characteristics during mountain-agarwood formation, and lay the foundation for exploring the mechanism of mountain-agarwood formation.

Morfoanatomía e histoquímica de Hypericum juniperinum (Hypericaceae)

Hypericum es un género que pertenece a la familia Hypericaceae, conformado por 469 especies de hierbas, árboles y arbustos. Dos secciones en particular, Brathys y Trygynobrathys, son parte integral de la flora de Páramo. Hypericum tiene una amplia variedad de metabolitos que están asociados con múltiples actividades farmacológicas, y específicamente en H. juniperinum se han reportado propiedades antidepresivas relacionadas con la presencia de flavonoides y compuestos fenólicos. El objetivo de esta investigación fue relacionar las estructuras anatómicas con los metabolitos detectados por métodos histoquímicos presentes en los órganos vegetativos y reproductivos de Hypericum juniperinum recolectados en el Páramo colombiano. Los órganos aéreos (tallos, hojas y flores) de Hypericum juniperinum se recolectaron en la vereda Arbolocos del municipio de Cuitiva, Boyacá (código voucher No.589611, COL). Para el análisis anatómico, los órganos de H. juniperinum se procesaron con técnicas de fijación, incorporación en parafina, corte en microtomo y tinción con azul de astra y fucsina básica. Para el análisis histoquímico se realizaron cortes a mano alzada con tinciones específicas para lípidos, compuestos fenólicos, alcaloides y pectinas. La anatomía de los órganos de H. juniperinum debe entenderse en el contexto de las condiciones de vida del Páramo. Las hojas son altamente xeromorfas ya que son cartáceas, arrosetadas, convolutas y verticales, con parénquima en empalizada biestratificado, paredes engrosadas, cutícula gruesa y cera epicuticular en ambas epidermis foliares, estomas en el lado abaxial con cámara epidérmica. Sin embargo, tiene algunas características mesomórficas, como las vainas parenquimáticas de los haces vasculares y el parénquima esponjoso suelto. Hypericum juniperinum presenta glándulas translúcidas en hojas, cáliz y gineceo, y carece de las glándulas negras típicas de la sección Hypericum a la que pertenece la planta medicinal Hypericum perforatum. El análisis histoquímico mostró lípidos en los canales de la hoja, parénquima radial del tallo y en algunas estructuras florales. Los compuestos fenólicos se detectaron a lo largo de la hoja, en el floema del tallo y en las anteras. Hypericum juniperinum carece de alcaloides y presenta pectinas en el parénquima de la vaina que rodea el haz vascular de la hoja. El presente trabajo contribuye a relacionar los metabolitos con su distribución en los tejidos de esta especie nativa. Además, la anatomía mostró que H. juniperinum tiene características de una planta de Páramo. La descripción que se proporciona aquí podría utilizarse como una monografía preliminar de plantas del género Hypericum con propiedades terapéuticas prometedoras.

Hypericum is a genus that belongs to the Hypericaceae family, with has 469 species of herbs, trees and shrubs. Two sections in particular, Brathys and Trygynobrathys are an integral part of the Paramo flora. Hypericum genus has a wide variety of metabolites that are associated with multiple pharmacological activities, and specifically in H. juniperinum antidepressant properties related to the presence of flavonoids and phenolic compounds have been reported. The objective of this research is to relate the anatomical structures with the metabolites detected by histochemical methods present in the vegetative and reproductive organs of Hypericum juniperinum collected in the Colombian Paramo. The aerial organs (stems, leaves and flowers) of Hypericum juniperinum were collected in the village Arbolocos of the municipality of Cuitiva, Boyacá (voucher code No. 589611, COL). For the anatomical analysis, the organs of H. juniperinum were processed with fixation techniques, incorporation in paraffin, cutting in microtome and staining with Astra-blue and basic fuchsin; for the histochemical analysis freehand cuts were made with specific stains for lipids, phenol compounds, alkaloids and pectins. The anatomy of the organs of H. juniperinum should be understood in the context of the living conditions of the Paramo. Leaves are highly xeromorphic as they are chartaceous, rosette-shaped, convolute and upright, with parenchyma in bistratified palisade, thickened walls, thick cuticle and epicuticular wax in both foliar epidermis, stomata on the abaxial side and with epidermal chamber. However, it has some mesomorphic characteristics such as parenchymatic bundle sheath and loose spongy parenchyma. Hypericum juniperinum has translucent glands in leaves, calyx and gynoecium, and lacks black glands typical of the Hypericum section belonging to the medicinal plant Hypericum perforatum. Histochemical analysis showed lipids in leaf canals, radial parenchyma of the stem and in some floral structures. Phenolic compounds were detected along the leaf, in the phloem of the stem and in the anthers. Hypericum juniperinum lacks alkaloids and presents pectins in the parenchyma of the sheath surrounding the vascular bundle of the leaf. The present work contributes by relating the metabolites with their distribution in the tissues of this native species. In addition, the anatomy of H. juniperinum has characteristics of the Paramo plants. Description given here could be used as a preliminary monograph of plants from the Hypericum genus with promising therapeutic properties.

Investigation of the effect of GnRH (deslorelin) on the histochemical structure of salivary glands in rats / Investigación del efecto de la GnRH (deslorelina) sobre la estructura histoquímica de las glándulas salivales en ratas

This study was planned to determine the histochemical alterations of the submandibular gland by implantation of long-term GnRH (deslorelin 4.7 mg). Eighteen Wistar albino rats were used in the study. Alcian blue (AB; pH: 2.5), periodic acid-Schiff (PAS) staining was performed to determine the microscopic structure and histochemical structure of the GI submandibular gland. The Avidin-Biotin Complex (ABC) method was used to determine the immunohistochemical reactivity of lectin. After GnRH implantation, the organs were examined and atrophies were observed in organs. In the group in which the implants were removed, it was determined that there was no atrophy; organ structures and microscopic examination were similar to the control group. At the end of the study, submandibular gland was fixed in 10 % buffered formaldehyde. In three groups, PAS and AB histochemical staining revealed similar reactions. Immunohistochemically, lectin activity was found to react positively.

Este estudio se planificó para determinar las alteraciones histoquímicas de la glándula submandibular mediante la implantación de GnRH a largo plazo (deslorelina 4,7 mg). Dieciocho ratas Wistar albinas se utilizaron en el estudio. Para determinar la estructura microscópica e histoquímica de la glándula submandibular, se realizó una tinción con azul alcián (AA; pH: 2.5) y ácido peryódico de Schiff (PAS). El método Avidin-Biotin Complex (ABC) se utilizó para determinar la reactividad inmunohistoquímica de la lectina. Después de la implantación de GnRH, se examinaron los órganos y se observó atrofia en ellos. En el grupo en el que se retiraron los implantes, no se observó atrofia. Las estructuras orgánicas y el examen microscópico fueron similares al grupo control. Al final del estudio, la glándula submandibular se fijó en formaldehído tamponado al 10 %. En tres grupos, la tinción histoquímica de PAS y AA reveló reacciones simila4res. Inmunohisto-químicamente, se encontró que la actividad de la lectina reaccionó positivamente.

Clinical-epidemiological, anatomic-pathological, histochemical and immunohistochemical characterization of renal cystadenocarcinoma-nodular dermatofibrosis syndrome in 11 German Shepherd dogs / Caracterização clínico-epidemiológica, anátomo-patológica, histoquímica e imuno-histoquímica da síndrome cistadenocarcinoma-dermatofibrose nodular em 11 cães Pastor Alemão

Eleven cases of renal cystadenoma/cystadenocarcinoma-nodular dermatofibrosis syndrome (RCND) are described in German Shepherd dogs diagnosed from January 1994 to January 2018 at the Veterinary Pathology Laboratory of the "Universidade Federal de Santa Maria" (LPV-UFSM). The study sample was composed of eight male and three female dogs at a ratio of 2.67:1. Age ranged from six to 12 years (mean=8.7 years). The main clinical signs reported in descending order of frequency were multiple cutaneous nodules (nodular dermatofibrosis), dyspnea, anorexia, weight loss, recurrent hematuria, vomiting, and polydipsia. Results demonstrated that it is not always easy to clinically recognize this syndrome, but its peculiar anatomical-pathological characteristics allow safe diagnosis. Histologically, it was possible to detect all phases (cysts, papillary intratubular hyperplasia, and cystadenomas or cystadenocarcinomas) of a possible pathological continuum of the renal lesions. Uterine leiomyomas were observed in only one of the cases. Through histochemical techniques, it was possible to identify the presence of type I collagen in both cutaneous and renal lesions and consider its possible involvement in the pathogenesis of renal cystadenocarcinoma. Immunohistochemistry (IHC) showed partially satisfactory results in the staining of epithelial cells of renal cysts and neoplasms for pan-cytokeratin.(AU)

São descritos 11 casos da síndrome cistadenoma/cistadenocarcinoma-dermatofibrose nodular (CR-DN) em cães Pastor Alemão, diagnosticados entre janeiro de 1994 e janeiro de 2018 no Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM). Os cães afetados foram oito machos e três fêmeas, estabelecendo-se uma relação de 2,67:1. A idade variou de seis a 12 anos, sendo a média de idade de 8,7 anos. Os principais sinais clínicos relatados foram, em ordem decrescente de frequência, múltiplos nódulos cutâneos (dermatofibrose nodular), dispneia, anorexia, emagrecimento, hematúria recorrente, vômito e polidipsia. Este estudo permitiu estabelecer que o reconhecimento clínico da síndrome nem sempre é fácil, porém suas características anátomo-patológicas peculiares permitem um diagnóstico com segurança. Histologicamente, foi possível detectar todas as fases (cistos, hiperplasia intratubular papilífera, cistadenomas ou cistadenocarcinomas) de um possível continuum patológico das lesões renais. Leiomiomas uterinos foram observados somente em um caso. Através das técnicas histoquímicas foi possível estabelecer que o colágeno tipo I está presente em ambas as lesões, cutâneas e renais, e cogitar seu possível envolvimento na patogênese dos cistadenocarcinomas renais. A técnica de IHQ mostrou resultados parcialmente satisfatórios na imunomarcação das células epiteliais dos cistos e dos neoplasmas renais para pancitoceratina.(AU)

Gastrointestinal Stromal Tumor (GIST)-Clinical Profile & Management of Patients Admitted To a Tertiary Care Hospital in Odisha

Gastrointestinal stromal tumor (GIST) is the commonest mesenchymal tumor of gastrointestinal tract. Gastrointestinal bleeding, obstruction, pain and abdominal lump are the common clinical manifestations. Local or segmental resection provides satisfactory results. Aim: Our aim was to report our experience of gastrointestinal stromal tumors (GISTs) during the last 2 years. Methods: Between January 2017 and June 2019, we performed surgery for 12 cases of GIST. Metastases, recurrence and survival data were collected in relation to age, history, clinical presentation, location, size, resection margins and cellular features. Results: Resection was completed in 11 cases. In one case definitive surgery was abandoned due to local invasion and metastasis. Three patients with high risk GIST were treated with imatinib mesylate. Conclusion: Non-radical surgery in the form of local or segmental resection is the standard surgical approach for GIST management. Pathological and biological features of the neoplasm represent the most important factors predicting the prognosis.