Histomorphological, Histochemical and Ultrastructural Studies on the Healthy Liver of Yemen Veiled Chameleon (Chamaeleo calyptratus) in Southern Saudi Arabia / Estudios Histomorfológicos, Histoquímicos y Ultraestructurales en el Hígado Sano del Camaleón Velado de Yemen (Chamaeleo calyptratus) en el Sur de Arabia Saudita

SUMMARY: The livers of reptiles are being studied as a model for the link between the environment and hepatic tissue. There have been few investigations on the histology of reptile livers, and very few or no studies have examined the histology of liver of veiled chameleon (Chamaeleo calyptratus). This paper describes the histomorphological, histochemical and ultrastructural characterization of the liver of veiled chameleons in southern Saudi Arabia. Seven Chamaeleo calyptratus were captured in the summer season in Abha City, Aseer region, southern Saudi Arabia. Chamaeleon liver samples were processed for histomorphology, histochemistry and ultrastructure analyses. Morphologically liver of Chamaeleo calyptratus was observed as a large dark brown organ with lighter speckles, which represent melanin deposits. It located at the ventral part of abdominal cavity forward of the stomach. Its dimensions approximately were 3.7 x 2 cm. The liver was a bilobed organ divided into two lobes, right and left lobes. The right one was bigger than the others. The gallbladder was well developed and had an elongated shape, situated between the two lobes and contained the bile for the digestion. Microscopically, the liver was found to be covered by a thick layer of connective tissue, which formed the hepatic capsule. Hepatic parenchyma probably appeared in cross sections as hepatic glandular-like alveoli "acini" or follicular structures with various diameters, each acinus contains approximately four to six hepatocytes, surrounded by sinusoidal capillaries filled with abundant melanomacrophages, which are absent in birds and mammals. Melanomacrophages are common in the hepatic parenchyma's perisinusoidal areas, particularly near portal spaces. Hepatocytes are polyhedral or pyramidal with and mostly contained large, rounded nuclei mostly peripherally located, with prominent dark oval nucleoli. Some of nuclei are eccentric or central position. The cytoplasm appeared spongy or vacuolated and more eosinophilic when stained by hematoxylin-eosin and strongly reactive to PAS staining technique, indicating abundant glycogen content. The reticular fibers that surround hepatocytes, blood arteries, and sinusoids supported the hepatic parenchyma. The blood sinusoids are seen interspersed among hepatocytes of varying sizes. The sinusoidal lumen was bordered by flattened endothelial cells and includes elliptical nucleated erythrocytes and liver macrophages as phagocytes, which are also known as Kupffer cells. Branches of the portal vein, hepatic artery, small bile duct, and lymph vessels were detected in the hepatic portal area "tract" or triad which made up of connective. Hematopoietic tissue was observed in subcapsular region and portal triads. Ultrastructurally, the hepatocyte appeared polyhedric containing a single large rounded basal or eccentric vesicular nucleus with prominent nucleolus. Extensive network of rough endoplasmic reticulum (RER) often arranged in an array parallel to the nuclear membrane with many mitochondria, and Golgi apparatus were described. The cytoplasm contained glycogen granules, vesicles or vacuoles scattered throughout the cytoplasm especially at the apical region were reported. The bile canaliculi and the hepatic "Kupffer" cells were also discussed. This is the first study on the histological characterization of the healthy liver of Yemen veiled chameleon in southern Saudi Arabia. The findings reported here should be used as a reference to compare with the pathological abnormalities of the liver in this animal.

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A morphological, histological and histochemical study of the sexual segment of the kidney of the male Chamaeleo calyptratus (veiled chameleon) / Un estudio morfológico, histológico e histoquímico del segmento sexual del riñón del macho Chamaeleo calyptratus (camaleón velado)

SUMMARY: This study aimed to describe the morphology, histology, and histochemistry of the kidneys and the sexual segment of the kidney (SSK) in Chamaeleo calyptratus (Yemen chameleons or veiled chameleons) collected during the active reproductive season. To achieve this objective, a total of 7 mature male Chamaeleo calyptratus were captured from their natural habitats from different areas in the Aseer region of Saudi Arabia and transferred to the reptilian laboratory where their kidneys were dissected out. Next, a morphological examination was conducted on the kidneys and the SSK was processed for light microscopic examinations with the primary focus placed on the morphology and histology of the SSK. The results of the study showed that Chamaeleo calyptratus possess two, reddish-brown, attenuated kidneys divided into lobes and lobules (lobulated). The histological study showed that the chameleons had no distinct limits between the cortex and medulla of the kidneys. In general, similar with other reptiles, this lizard demonstrated that the kidneys have few nephrons, do not show any nephron loop (loop of Henle), and display few glomeruli. The nephron is generally composed of standard components: renal corpuscle, glomerulus surrounded by a double-walled, the glomerular capsule (Bowman's capsule) and renal tubules (six renal tubular segments differentiated as the ciliated neck segment [between the glomerulus and the proximal segment], proximal segment, ciliated intermediate segment instead of nephron loop that connects proximal and distal, distal segment, terminal segment or sexual segment, and all segments emptying into the collecting ducts. The epithelial tissues lining the lumen of these segments vary from simple cuboidal cells to columnar and tall columnar cells in the SSK. Mature male Chamaeleo calyptratus possess hypertrophied parts of the distal renal segments, which are called the SSK. Histologically, the active SSK consists of simple tall columnar epithelial cells with rounded basal nuclei with clear centralized nucleoli and numerous apical coarse secretory granules which are released from the cells by an apocrine process. Histochemically, the secretory granules stain positively for proteins with mercuric bromophenol blue and react intensely to periodic acid-Schiff (PAS) for neutral carbohydrates. This is the first report about the histology of the SSK of the Chamaeleo calyptratus, and this study adds to the knowledge of the reproductive biology, biodiversity, and reproductive strategies of chameleons. Further studies are required to investigate the ultrastructure of the kidneys and SSK.

RESUMEN: Este estudio tuvo como objetivo describir la morfología, histología e histoquímica de los riñones y el segmento sexual en el riñón (SSK) del Chamaeleo calyptratus (camaleones de Yemen o camaleones con velo) recolectados durante la temporada reproductiva. Un total de 7 machos maduros de Chamaeleo calyptratus fueron capturados en su hábitat natural de diferentes áreas en la región de Aseer de Arabia Saudita y transferidos al laboratorio de reptiles donde fueron disecados los riñones. Posteriormente, se llevó a cabo un examen morfológico de los riñones y se procesó la SSK para exámenes con microscopio óptico con el enfoque principal puesto en la morfología y la histología. Los resultados del estudio mostraron que Chamaeleo calyptratus posee dos riñones atenuados de color marrón rojizo divididos en lóbulos (lobulados). En el estudio histológico se observó que los riñones de los camaleones no tenían límites definidos entre la corteza y la médula. En general, y similar a otros reptiles, este lagarto demostró que los riñones tienen pocas nefronas, no muestran ningún asa de Henle y presentan pocos glomérulos. La nefrona está compuesta por componentes estándar: corpúsculo renal (glomérulo rodeado por una pared de doble pared). Cápsula de Bowman y túbulos renales (seis segmentos tubulares renales diferenciados como el segmento del cuello ciliado [entre el glomérulo y el segmento proximal], segmento proximal, segmento intermedio ciliado en lugar del asa de Henle que conecta proximal y distal, segmento distal, segmento terminal o segmento sexual y todos los segmentos que desembocan en los conductos colectores Los tejidos epiteliales que recubren la luz de estos segmentos varían desde células cuboidales simples hasta células columnares y columnares altas en la SSK. Los machos maduros de Chamaeleo calyptratus poseen partes hipertrofiadas de los segmentos renales distales, denominados SSK. Histológicamente, la SSK activa consiste en células epiteliales cilíndricas altas simples con núcleos basales redondeados, nucléolos centralizados claros y gránulos secretores gruesos apicales que se liberan de las células por un proceso apocrino. Histoquímicamente, los gránulos secretores se tiñen positivamente para las proteínas con azul de bromofenol mercúrico y reaccionan intensamente al ácido periódico-Schiff (PAS) para obtener carbohidratos neutros. Este es el primer informe sobre la histología de la SSK de Chamaeleo calyptratus, y se suma al conocimiento de la biología reproductiva, la biodiversidad y las estrategias reproductivas de los camaleones. Se requieren más estudios para investigar la ultraestructura de los riñones y la SSK.

The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats / O óleo de alho, Allium sativum L. (Amaryllidaceae), como potencial protetor contra Anisakis spp. Infecção tipo II (L3) (Nematoda) em ratos Wistar

Abstract The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Resumo O consumo de peixe inadequadamente tratado termicamente representa um risco para a saúde pública, com a possibilidade da propagação de larvas de Anisakis. O presente estudo demonstrou as alterações fisiológicas e histopatológicas acompanhadas de inoculação oral de extractos brutos de Anisakis tipo II (L3) frescos e termicamente tratados em ratos. Os vermes foram isolados de um peixe marinho, examinados e identificados por microscopia de luz e eletrônica de varredura. O estudo foi conduzido em 6 grupos de ratos: controle (I), óleo de alho (GO) inoculado (II), L3 fresco inoculado (III), L3 tratado termicamente inoculado (IV), L3 fresco + GO inoculado (V), e um grupo L3 + GO tratado termicamente inoculado (VI). Observou-se que ratos inoculados com L3 fresco e tratados termicamente mostraram funções hepáticas e renais anormais, associadas à destruição da sua arquitetura normal. GO produziu um efeito protector em grupos de ratos inoculados com extractos L3 + GO através da melhoria das funções do fígado e dos rins, o que foi confirmado pela estrutura normal marcada da sua histologia. A cozedura de peixes infectados com L3 induziu alterações mais graves do que os peixes não cozidos. Recomenda-se a administração de alho antes e depois do consumo de peixe, para evitar o efeito perigoso dos anisakids, mesmo que sejam cozidos.

Hepatotoxicity-induced by the therapeutic dose of acetaminophen and the ameliorative effect of oral co-administration of selenium/Tribulus terrestris extract in rats / Hepatotoxicidad inducida por la dosis terapéutica de acetaminofén y el efecto de mejora de la administración conjunta de extracto de selenio Tribulus terrestris en ratas

SUMMARY: Over dose or long-term clinical use of therapeutic doses of acetaminophen (APAP) causes hepatotoxicity. Various strategies attempted to ameliorate APAP-hepatotoxicity have been found to be unsuitable for clinical practice. This study was aimed to illustrate the histopathological changes induced by therapeutic dose of APAP and investigate the hepatoprotective role of oral co-administration of selenium/ Tribulus terrestris (TT) extract concurrently against hepatotoxicity induced by APAP in rats. Fifty-four healthy male albino Wistar rats were randomized into nine groups (G1-G9) of six rats each, and administered with APAP and TT orally for 30 days as follows: Control (2ml normal saline), APAP (470 mg/kg), APAP (470 mg/kg) + selenium (2 mg/kg), APAP (470 mg/kg) + TT (98 mg/kg), APAP (470 mg/kg) + selenium (2mg/kg) + TT (98 mg/kg), APAP (470 mg/kg) + silymarin (200 mg/kg), selenium (2 mg/ kg), TT (98 mg/kg) and silymarin (200 mg/kg) groups. The results demonstrated that exposure of rats to therapeutic dose of APAP for 30 days caused significant histopathological changes parallel to elevated blood chemistry parameters. Co-administration of selenium/TT extract showed significantly reduced histopathological lesions and, restored or decreased levels of the examined blood chemistry parameters. Liver histology in selenium/TT extract showed normal hepatic architecture with mild changes and silymarin treated rats showed no histopathological changes. Histochemically PAS staining, showed that APAP-induced hepatotoxicity was characterized by hepatocytes glycogen depletion. Selenium/TT co-supplementation plays a potential role in preventing APAP-induced glycogen depletion by increasing detoxification and scavenging the reactive metabolites. Selenium/TT extract oral co-administration possesses a significant hepatoprotective property and mitigates APAP-induced hepatotoxicity by enhancing its antioxidant role and improving tissue integrity. Selenium/TT supplementation could represent an effective treatment against APAP-induced hepatotoxicity. Further studies are needed to elucidate the exact mechanism underlying the protective role of TT extract.

RESUMEN: La dosis excesiva o el uso clínico a largo plazo de dosis terapéuticas de acetaminofeno (APAP) causa hepatotoxicidad. Se ha descubierto que varias estrategias que intentaron mejorar la hepatotoxicidad por APAP no son adecuadas para la práctica clínica. Este estudio tuvo como objetivo ilustrar los cambios histopatológicos inducidos por la dosis terapéutica de APAP e investigar el papel hepatoprotector de la administración conjunta de extracto de selenio / Tribulus terrestris (TT) simultá- neamente contra la hepatotoxicidad inducida por APAP en ratas. Cincuenta y cuatro ratas Wistar albino machos sanas se aleatorizaron en nueve grupos (G1 - G9) de seis ratas cada una, y se administraron con APAP y TT por vía oral durante 30 días de la siguiente manera: Control (2 ml de solución salina normal), APAP (470 mg / kg), APAP (470 mg / kg) + selenio (2 mg / kg), APAP (470 mg / kg) + TT (98 mg / kg), APAP (470 mg / kg) + selenio (2 mg / kg) + TT (98 mg / kg), APAP (470 mg / kg) + silimarina (200 mg / kg), selenio (2 mg / kg), TT (98 mg / kg) y silimarina (200 mg / kg). Los resultados demostraron que la exposición de las ratas a la dosis terapéutica de APAP durante 30 días causó cambios histopatológicos significativos paralelos a parámetros elevados de química sanguínea. La administración conjunta de extracto de selenio / TT mostró lesiones histopatológicas significativamente reducidas y niveles restaurados o disminuidos de los parámetros de química sanguínea. La histología hepática en el extracto de selenio / TT mostró una arquitectura hepática normal con cambios leves y las ratas tratadas con silimarina no mostraron cambios histopatológicos. La tinción histoquímica de PAS mostró que la hepatotoxicidad inducida por APAP se caracterizó por la pérdida de glucógeno de los hepatocitos. La suplementación con selenio / TT juega un papel potencial en la prevención de la pérdida de glucógeno inducido por APAP al aumentar la desintoxicación y eliminar los metabolitos reactivos. La administración conjunta de extracto de selenio / TT posee una propiedad hepatoprotectora significativa y mitiga la hepatotoxicidad inducida por APAP al mejorar su papel antioxidante y la integridad del tejido. La suplementación con selenio / TT podría representar un tratamiento efectivo contra la hepatotoxicidad inducida por APAP. Se necesitan más estudios para dilucidar el mecanismo exacto que subyace a la función protectora del extracto TT.

Protective effect of gum arabic/insulin against histological changes in testes of diabetic rats / Efecto protector de la goma arábiga / insulina contra los cambios histológicos en los testículos de ratas diabéticas

Diabetes mellitus is a serious disease with a high incidence of occurrence in our community. Gum Arabic (GA) is a branched-chain polysaccharide which has strong antioxidant properties, and has been used to reduce the experimental toxicity. Yet, the effects of GA on testicular tissue in type I diabetic rats have not been enough investigated. This study was designed to investigate histological changes in testes of male Wistar rats and investigate the protective potential of GA against diabetes- induced testicular toxicity in rats. Fifty adult male Wistar rats were assigned into five groups (n = 10 of each): Group 1 (non-diabetic rats) served as control, Group 2 served as diabetic group injected with Alloxan, Group 3 diabetic group plus insulin, Group 4 diabetic group given 15 % GA in drinking water and Group 5 diabetic group plus insulin and GA for 4 weeks. Compared to control group, histopathological examinations of testicular tissue from the diabetic rats group, showed degeneration, necrosis and atrophy of seminiferous with presence of giant cells. Necrosis and hemorrhage in the renal tissue. On the other hand, treatment with GA ameliorated all the previous histological changes. Overall, oral administration of GA alone or with insulin daily for 4 weeks successfully ameliorated the testicular histological changes. These data demonstrated that GA significantly improved diabetes complication in rat testis. This study suggested that GA might have a protective effect against oxidative stress-induced impaired testicular functions in diabetic rats. The possible mechanisms of this action might be ascribed to their antioxidant and anti-inflammatory properties.

La diabetes mellitus es una enfermedad grave con una alta incidencia en nuestra comunidad. La goma arábiga (GA) es un polisacárido con propiedades antioxidantes importantes, y se ha utilizado para reducir la toxicidad experimental. Sin embargo, los efectos de GA sobre el tejido testicular en ratas diabéticas tipo I no se ha investigado lo suficiente. El estudio fue diseñado para pesquisar los cambios histológicos en los testículos de ratas Wistar macho e investigar el potencial protector de GA contra la toxicidad testicular inducida por la diabetes en ratas. Fueron asignadas cincuenta ratas Wistar macho adultas en cinco grupos (n = 10 de cada una): el grupo 1 (ratas no diabéticas) sirvió como control, el grupo 2 sirvió como grupo diabético inyectado con Alloxan, grupo diabético del grupo 3 más insulina. El grupo 4 diabético recibió 15 % de GA en agua potable, y el grupo diabético 5 más insulina y GA durante 4 semanas. Al comparar con el grupo control, los exámenes histopatológicos del tejido testicular del grupo de ratas diabéticas mostraron degeneración, necrosis y atrofia de los túbulos seminíferos con presencia de células gigantes, necrosis y hemorragia en el tejido renal. Por otra parte, el tratamiento con GA mejoró todos los cambios histológicos previos. En general, la administración oral de GA solamente, o con insulina diariamente durante 4 semanas mejoró los cambios histológicos testiculares. Estos datos demostraron que GA mejoró significativamente los efectos de la diabetes en testículos de rata. Este estudio sugiere que GA podría tener un efecto protector contra las funciones testiculares deterioradas, inducidas por el estrés oxidativo en ratas diabéticas. Los posibles mecanismos de esta acción podrían atribuirse a sus propiedades antioxidantes y antiinflamatorias.

Renal histological alterations induced by acute exposure of titanium dioxide nanoparticles / Alteraciones histológicas renales inducidas por la exposición aguda de nanopartículas de dióxido de titanio

SUMMARY: Titanium dioxide nanoparticles (TiO2 NPs) are widely used in many commercial products, nanomedicine, agriculture, personal care products, different industries and pharmaceutical preparations with potential risk in human health and the environment. The current work was conducted to investigate the renal damage that might be induced by the acute toxicity TiO2 NPs. A total of 40 healthy male adult Wistar albino rats (Rattus norvegicus) were exposed to TiO2 NPs (126, 252, 378 mg/kg bw) for 24 and 48 h. Fresh portions of the kidneys from each rat were processed for histological and histochemical alterations. In comparison with respective control rats, exposure to TiO2 NPs has marked the following glomerular, tubular and interstitial alterations including the followings: glomerular congestion, Bowman's capsule swelling and dilatation, inflamed glomeruli, renal tubules cloudy swelling, karyorrhexis, karyolysis, infiltration of inflammatory cells, congestion, necrosis, hydropic degeneration, dilatation and congestion of blood vessels, hyaline droplets and hyaline casts precipitation, interstitial edema and fibrosis. From the findings of the current work one may conclude that TiO2 NPs are capable of inducing kidney damage with more insulation in the cortex and the proximal convoluted tubules than the medulla and the distal ones respectively. In addition, it might be concluded that renal damage induced by these nanomaterials is dose and duration of exposure dependent. Further hematological, biochemical, immunohistochemical, and ultra-structural studies are recommended.

RESUMEN: Las nanopartículas de dióxido de titanio (TiO2 NP) se usan ampliamente en muchos productos comerciales, nanomedicina, agricultura, productos para el cuidado personal, diferentes industrias y preparaciones farmacéuticas con riesgo potencial para la salud humana y el medio ambiente. El trabajo actual se realizó para investigar el daño renal que podría ser inducido por la toxicidad aguda NP de TiO2. Un total de 40 ratas Wistar albinas adultas sanas (Rattus norvegicus) fueron expuestas a TiO2 NP (126, 252, 378 mg / kg de peso corporal) durante 24 y 48 h. Las muestras de los riñones de las ratas se procesaron para estudios histológicos e histoquímicos. En comparación con las ratas control, la exposición de las ratas a TiO2 NP presentaron las siguientes alteraciones glomerulares, tubulares e intersticiales: congestión glomerular, dilatación de la cápsula de Bowman, inflamación glomerular, túbulos renales aumentados, cariorrexis, cariólisis, infiltración de células inflamatorias, congestión, necrosis, degeneración hidrópica, dilatación y congestión de vasos sanguíneos, gotas y precipitaciones hialina, edema intersticial y fibrosis. A partir de los hallazgos del trabajo actual, se puede concluir que las NP de TiO 2 son capaces de inducir daño renal con más aislamiento en la corteza y en los túbulos contorneados proximales que en la médula y los túbulos contorneados distales, respectivamente. Además, se podría concluir que el daño renal inducido por estos nanomateriales depende de la dosis y la duración de la exposición. Se recomiendan estudios adicionales hematológicos, bioquímicos, inmunohistoquímicos y ultraestructurales.