Thick Basal Lamina Research Articles

ROLE OF KATHAKAKHADIRADI KASHAYA IN PREVENTIVE DIABETOLOGY; A CASE REPORT

Diabetes mellitus is an abnormal scenario of altered glucose tolerance due to inadequate insulin action. Small blood vessels are susceptible to the complications of diabetes mellitus, such as basal lamina thickening and endothelial cell proliferation called diabetic microangiopathy, frequently causing retinal and renal damage. Microangiopathy affecting the kidney is called diabetic nephropathy and is the second leading cause of death due to diabetes mellitus after myocardial infarction. Due to the persistent hyperglycemic status of microcapillaries in the kidney, some histological changes occur in the nephron level detected by electron microscopy only after a few years of onset of diabetes [1]. This is a case report of a known diabetic and hypertensive patient who complained of frothy urine and was diagnosed as type 2 diabetes-induced microalbuminuria and intervened with Kathaka Khadiradi Kashaya, a traditional antidiabetic formulation famous in South India. Two months of administration of the medicine reduced the urine microalbumin and albumin creatinine ratio to a significant normalcy, hence proved to be a choice for a clinical trial in the preventive aspect of diabetic nephropathy.

Effects of Meningic Extract on Testicular Torsion/Detorsion

Introduction: Testicular torsion leads to impaired blood supply to the testicles and ischaemia. The incidence is higher especially in newborns and adolescents. As in the treatment of many diseases, various plant combinations are used to prevent the damage caused by testicular torsion-detorsion. In this study, we aimed to investigate the protective effect of meningic (Pistacia terebinthus L.) extract on testicular torsion-detorsion (T/D) injury histopathologically.
 Materials and Methods: The 24 animals used in our experiment were randomly divided into 3 groups. Testicular tissues were stained with hematoxylin-eosin and PAS stains to evaluate histological changes. Modified Johnsen scoring was then performed.
 Results: In the sham group sections, seminiferous tubules were regular and germinal epithelium at various stages of spermatogenesis was seen in 4-5 rows on the basal lamina. In PAS stained sections of the sham group, the basement membrane was regular. In the sections of T/D group, destruction of seminiferous tubules, dilatation and congestion were observed in the vessels in the interstitial area. Thickening of the basal laminae of the seminiferous tubules was also detected in the sections of the T/D group stained with PAS. In the sections of meningic + T/D group, a partial recovery was observed in the seminiferous tubules. The basal lamina thickness in PAS-stained sections of meningic + T/D group was found to be close to normal. According to the results of modified Johnsen score analysis, there was a significant difference between the groups (p<0.05).
 Conclusion: In our study, it was concluded that meningih extract partially protects the testis and its functions.
 Keywords: Testicular injury, torsion, detorsion, meningic extract

The pecten oculi comparison of the different bird species

The pecten oculi is a unique and highly specialized structure found in the eyes of birds. It is a thin, folded, fan-like membrane that extends from the optic nerve region into the vitreous fluid of the eyeball. This membrane is primarily composed of pigment cells, an extensive network of blood vessels, and a thick basal lamina. The pecten's main function is to supply nutrients and oxygen to the retina, maintain the internal eye temperature, and support sharp vision, especially during activities such as migration and hunting. The number of folds in the pecten varies between diurnal and nocturnal birds, with diurnal birds generally having larger and more complex pecten structures. The pigmentation of the pecten, especially at the apical and peripheral regions, plays a role in protecting the blood vessels from UV radiation and oxygen radicals. Different bird species exhibit variations in the number, size, and thickness of pecten folds, reflecting their specific visual requirements. For example, species like the emu have a more primitive, pleated pecten, while birds like the seagull and quail have a greater number of folds and capillaries. Additionally, the presence of melanocytes and their interactions with capillaries and blood vessels contribute to the pecten's structural integrity. The study of the pecten oculi's histology reveals that it is essential for the bird's visual health and can be linked to their lifestyle and environmental conditions. The structure and size of the pecten are tailored to the bird's daily activities, with diurnal birds having larger and more complex pecten structures compared to their nocturnal counterparts. This specialized organ's morphology and vascularization are crucial for delivering nutrients to the retina and protecting it from UV radiation. In summary, the pecten oculi is a remarkable anatomical feature in the avian eye, and its structure and function are closely related to the lifestyle and visual requirements of different bird species. This study provides valuable insights into the diverse adaptations of the pecten across bird species and its significance in maintaining avian visual health.

Developmental and mechanical roles of Reichert's membrane in mouse embryos.

Embryonic development and growth in placental mammals proceeds in utero with the support of exchanges of gases, nutrients and waste products between maternal tissues and offspring. Murine embryos are surrounded by several extraembryonic membranes, parietal and visceral yolk sacs, and amnion in the uterus. Notably, the parietal yolk sac is the most outer membrane, consists of three layers, trophoblasts and parietal endoderm (PaE) cells, and is separated by a thick basal lamina termed Reichert's membrane (RM). RM is composed of extracellular matrix (ECM) initially formed as the basement membrane of the trophectoderm of pre-implanted embryos and followed by the heavy deposition of ECM mainly produced in PaE cells of post-implanted embryos. In addition to the physiological roles of RM, such as gas and nutrient exchange, it also plays a crucial role in cushioning and dispersing intrauterine pressures exerted on embryos for normal egg-cylinder morphogenesis. Mechanistically, such intrauterine pressures generated by uterine smooth muscle contractions appear to be involved in the elongation of the egg-cylinder shape, along with primary axis formation, as an important biomechanical element in utero. This review focuses on our current views of the roles of RM in properly buffering intrauterine mechanical forces for mouse egg-cylinder morphogenesis. This article is part of the theme issue 'Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom'.

Malaria in Pregnancy: From Placental Infection to Its Abnormal Development and Damage.

Malaria remains a global health burden with Plasmodium falciparum accounting for the highest mortality and morbidity. Malaria in pregnancy can lead to the development of placental malaria, where P. falciparum-infected erythrocytes adhere to placental receptors, triggering placental inflammation and subsequent damage, causing harm to both mother and her infant. Histopathological studies of P. falciparum-infected placentas revealed various placental abnormalities such as excessive perivillous fibrinoid deposits, breakdown of syncytiotrophoblast integrity, trophoblast basal lamina thickening, increased syncytial knotting, and accumulation of mononuclear immune cells within intervillous spaces. These events in turn, are likely to impair placental development and function, ultimately causing placental insufficiency, intrauterine growth restriction, preterm delivery and low birth weight. Hence, a better understanding of the mechanisms behind placental alterations and damage during placental malaria is needed for the design of effective interventions. In this review, using evidence from human studies and murine models, an integrated view on the potential mechanisms underlying placental pathologies in malaria in pregnancy is provided. The molecular, immunological and metabolic changes in infected placentas that reflect their responses to the parasitic infection and injury are discussed. Finally, potential models that can be used by researchers to improve our understanding on the pathogenesis of malaria in pregnancy and placental pathologies are presented.

HISTOLOGICAL OF CHANGES IN THE AMNIOTIC MEMBRANE AND PLACENTAL VILLOUS BASAL LAMINA IN COMPLICATED PREGNANCIES

Aim: This study aimed to histological changes compare placental villous basal lamina and amniotic membrane changes in complicated pregnancy. Methods : Studies were performed on the human placentas, delivered between 24-39 weeks of gestation. Patients were separated equally into 4 groups ( Control, preeclampsia (PE), gestastional diabetes (GD), and HELLP syndrome groups ) Placental tissue samples were dissected and fixed in 10% neutral formalin buffer. Routine paraffin tissue protocol was followed. Some of sections were stained with Periodic Acid Schiff. Remaining sections were stained with integrin alpha-6 antibody.To define expression percentage, mean of the staining area/total staining area ratio were calculated. The statistical significance of the expression percentages was compared by One Way ANOVA and Tukey tests with SPSS Statistics V24 software. Results: In PAS-stained preeclamptic, HELLP and gestational diabetes groups placental villous basal lamina and vasculo-syncytial membranes were thicker than the control group. A significant difference was observed in all 3 groups compared to the control group the placental villous basal lamina thickness of the HELLP group was found to be significantly different from all three groups. In chorionic villi of HELLP group, dense integrin expression was found in placental villous basal lamina similar to that in GD and preeclampsia groups. The HELLP group was significantly different from all groups. Conclusion: In preeclampsia, gestational diabetes and HELLP placentas, placental viioz basal lamina and amniotic membrane significantly thickened and structural changes were observed.

Omalizumab may decrease the thickness of the reticular basement membrane and fibronectin deposit in the bronchial mucosa of severe allergic asthmatics

Introduction: Immunoglobulin E is an important modulator of the inflammatory reaction in allergic asthma. It also contributes to airway remodeling in the course of the disease. The authors evaluated airway structural changes in severe allergic asthma during the omalizumab therapy. Patients and methods: The study included 13 patients with severe allergic asthma treated with omalizumab for at least one year. In each patient clinical, laboratory, and spirometry parameters were evaluated before and after the treatment. In addition, bronchoscopy with bronchial mucosa biopsy and bronchoalveolar lavage was performed. The basal lamina thickness, inflammatory cell infiltration, fibronectin, as well as type I and III collagen accumulation were assessed in bronchial mucosa specimens, together with the assessment of bronchoalveolar lavage cellularity. Results: The omalizumab therapy led to a decrease in the basal lamina thickness (p = 0.002), and to a reduction in fibronectin (p = 0.02), but not collagen deposits in the bronchial mucosa. The decrease in fibronectin accumulation was associated with an improvement in asthma control and quality of life (p = 0.01, both), and a diminished dose of systemic corticosteroids (p = 0.001). It was also associated with a tendency towards reduction of the eosinophil count in the peripheral blood, bronchoalveolar lavage fluid, and bronchial mucosa specimens. Conclusion: Our study has shown that omalizumab, effective in the treatment of severe allergic asthma, may also decrease unfavorable structural airway changes in allergic asthmatics, at least with respect to the fibronectin deposit and an increased thickness of the basal lamina. However, more extensive observational studies are needed to verify the above hypothesis.

Basement membranes in the kidney of the dromedary camel (Camelus dromedarius): An immunohistochemical and ultrastructural study

Basement membranes consist of various proteins, the major ones being laminin and collagen type IV. Primary defects in these two proteins have been extensively associated with kidney pathologies. This study aimed to establish baseline information on the immunohistochemical distribution of laminin and collagen type IV, and to corelate these with the ultrastructure of basal laminae in the uriniferous tubules of the dromedary camel. Tissue samples were taken from the kidneys of eight adult female camels, and processed for immunohistochemical and ultrastructural investigations. Strong intensity of collagen type IV was observed within the basement membranes of Bowman’s capsule. The thickness of the basal lamina of the parietal layer of Bowman's capsule varied extensively depending on the region of the renal corpuscle; the thicker areas were always associated with cuboidal epithelial cells. The glomerular basement membrane revealed strong immunostaining of laminin, whereas the mesangial matrix was strongly immunoreactive to collagen type IV. Abundant amount of laminin was found in the basement membranes of proximal convoluted tubules, thin limbs of the loop of Henle, and collecting ducts. Dense immunostainings of laminin and collagen type IV were observed in the medullary regions of uriniferous tubule, in which numerous projections extended from the basal laminae into the subjacent connective tissue. Overall, the present study revealed marked variations in the distribution of the basement membrane markers laminin and collagen type IV in the uriniferous tubules of camel kidney. The results have also shown difference in the thickness of basal laminae. This variation in thickness, however, was unlikely to be influenced by the amount of laminin and collagen type IV.

Phosphodiesterase-5 Inhibition Alleviates Pulmonary Hypertension and Basal Lamina Thickening in Rats Challenged by Chronic Hypoxia.

Background: Hypoxia represents both an outcome of cardiopulmonary diseases and a trigger for severe pulmonary complications as pulmonary hypertension. Because nitric oxide (NO) is a critical mediator in the development of pulmonary hypertension, the modulators of its downstream function may become target of pharmacological interventions aimed at alleviating the impact of this condition. Here, we investigate the effects of an early administration of phosphodiesterase-5 inhibitor in rats where pulmonary artery hypertension was induced by chronic exposure to hypoxia.Methods: Rats were divided into three groups: normoxic control, hypoxic with no treatments (2 weeks breathing an atmosphere containing 10% oxygen), and hypoxic treated with sildenafil (1.4 mg/Kg per day in 0.3 mL i.p.). After sacrifice, hearts and lungs were removed and harvested for analyses.Results: Sildenafil reduced hypoxia-induced right ventricle hypertrophy without effects in lung hypertrophy, and blunted the increase in right ventricle pressure without effects on left ventricle pressure. Furthermore, the NO-producing systems (i.e., the phosphorylation of the endothelial isoforms of NO synthase that was measured in both myocardial and lung tissues), and the blood NO stores (i.e., the plasma level of nitrates and nitrites) were up-regulated by sildenafil. We did not find significant effects of sildenafil on weight and hemoglobin concentration. Morphological analysis in lung biopsies revealed that 2-week hypoxia increased the frequency of small pulmonary vessels leaving large vessels unaffected. Finally, ultrastructural analysis showed that sildenafil down-regulated the hypoxia-induced increase in the thickness of the pulmonary basal lamina.Conclusions: In this model of pulmonary hypertension, sildenafil contrasts the negative effects of hypoxia on pulmonary vascular and right ventricle remodeling. This action does not only encompass the canonical vasomodulatory effect, but involves several biochemical pathways. Although the human pathological model is certainly more complex than that described here (for example, the inflammatory issue), the potential role of phosphodiesterase-5 for long-term treatment, and perhaps prevention, of pulmonary hypertension is worthy of investigation.

Müller glial cells of the primate foveola: An electron microscopical study

Previous studies on the ultrastructure of the primate foveola suggested the presence of an inverted cone-like structure which is formed by 25–35 specialized Müller cells overlying the area of high photoreceptor density. We investigated the ultrastructure of the Müller cells in the foveola of a human and macaque retina. Sections through the posterior poles of an eye of a 40 years-old human donor and an eye of an adult cynomolgus monkey (Macaca fascicularis) were investigated with transmission electron microscopy. The foveola consisted of an inner layer (thickness, 5.5–12 μm) which mainly contained somata (including nuclei) and inner processes of Müller cells; this layer overlaid the central Henle fibers and outer nuclear layer. The inner layer contained numerous watery cysts and thin lamelliform and tubular Müller cell processes which spread along the inner limiting membrane (ILM). The cytoplasm of the outer Müller cell processes became increasingly dispersed and electron-lucent in the course towards the outer limiting membrane. The ILM of the foveola was formed by a very thin basal lamina (thickness, <40 nm) while the basal lamina of the parafovea was thick (0.9–1 μm). The data show that there are various conspicuous features of foveolar Müller cells. The numerous thin Müller cell processes below the ILM may smooth the inner surface of the foveola (to minimize image distortion resulting from varying light refraction angles at an uneven retinal surface), create additional barriers to the vitreous cavity (compensating the thinness of the ILM), and provide mechanical stability to the tissue. The decreasing density of the outer process cytoplasm may support the optical function of the foveola.

Metabolic Syndrome, Neurotoxic 1-Deoxysphingolipids and Nervous Tissue Inflammation in Chronic Idiopathic Axonal Polyneuropathy (CIAP).

AimChronic idiopathic axonal polyneuropathy (CIAP) is a slowly progressive, predominantly sensory, axonal polyneuropathy, with no aetiology being identified despite extensive investigations. We studied the potential role of the metabolic syndrome, neurotoxic 1-deoxysphingolipids (1-deoxySLs), microangiopathy and inflammation in sural nerve biopsies.MethodsWe included 30 CIAP-patients, 28 with diabetic distal symmetrical polyneuropathy (DSPN) and 31 healthy controls. We assessed standardised scales, tested for the metabolic syndrome, measured 1-deoxySLs in plasma, performed electroneurography and studied 17 sural nerve biopsies (10 CIAP; 7 DSPN).ResultsOne third of the CIAP-patients had a metabolic syndrome, significantly less frequent than DSPN-patients (89%). Although the metabolic syndrome was not significantly more prevalent in CIAP compared to healthy controls, hypercholesterolemia did occur significantly more frequent. 1-deoxySLs were significantly and equally elevated in both patient groups compared to healthy controls. Mean basal lamina thickness of small endoneurial vessels and the number of CD68- or CD8-positive cells in biopsies of CIAP- and DSPN-patients did not differ significantly. However, the number of leucocyte-common-antigen positive cells was significantly increased in CIAP.ConclusionsA non-significant trend towards a higher occurrence of the metabolic syndrome in CIAP-patients compared to healthy controls was found. 1-deoxySLs were significantly increased in plasma of CIAP-patients. Microangiopathy and an inflammatory component were present in CIAP-biopsies.

Vitiligo – The story from within: A transmission electron microscopic study before and after narrow-band ultraviolet B

ABSTRACTMelanocyte loss is the main feature of vitiligo, but evidence refers to pathological multiplayers. Transmission electron microscopy was utilized to further explore vitiligo before and after narrow-band ultraviolet B (NB-UVB) therapy. Skin biopsies were retrieved from lesional and perilesional skin and compared to normal control skin. Sections were examined for melanocytes and keratinocytes and the number of melanosomes and thickness of basal lamina were measured. In lesional skin, keratinocytes revealed two types of degeneration with a significant increase in the mean thickness of basal lamina and decrease in the number of melanosomes. After treatment, lesional and perilesional skin showed variable ultrastructural features.

Surface histology, topography, and ultrastructure of the tegument of adult Orthocoelium parvipapillatum (Stiles & Goldberger, 1910).

Adult Orthocoelium parvipapillatum are common parasites that reside in the rumen and reticulum of ruminants, i.e., cattle, sheep, goats, and buffaloes. The fluke is conical-shaped and slightly concave ventrally and convex dorsally, and measures bout 2.4-3.9mm in length and 1.0-2.3mm in width across the mid-section. The tegument of the adult worm is examined using light microscopy (LM) and scanning (SEM) and transmission electron microscopy (TEM). Under LM, the tegument appears as a thick homogeneous layer containing folds alternated with grooves without spines. SEM revealed that the tegumental surface is highly corrugated with ridges and furrows and appears spineless. Two types of sensory papillae are observed, i.e., type 1 is bulbous in shape with nipple-like tips and type 2 has a similar shape with short cilia. In TEM, the tegument has a typical syncytial organization and is divided into four layers. The first layer of the tegument contains ridges and furrows covered by a trilaminate membrane coated externally with the glycocalyx. The second layer is a strait area of cytoplasm that includes numerous ovoid electron-lucent (TG1) and disc-shaped electron-dense (TG2) tegumental granules and lysosomes. The third layer is the widest middle area which contains several evenly distributed mitochondria, TG1 and TG2. The fourth layer rests on a thick basal lamina and contains numerous infoldings of the basal plasma membrane with closely associated mitochondria. Both granules are produced and transported to the tegument by one type of tegumental cells lying in rows below the muscular layers.

Anatomy and Ultrastructure of Male Reproductive System in Two LeafhoppersCicadella viridisandKolla paulula(Hemiptera: Auchenorrhyncha: Cicadellidae)

The morphology and ultrastructure of the male reproductive system were investigated in two species of sharpshooter leafhoppers, Cicadella viridis (L.) and Kolla paulula (Walker), using light microscopy and transmission electron microscopy. The gross morphology and ultrastructure of the male reproductive system in the two species is similar to those of other Cicadellidae and related Auchenorrhyncha. The two species differ in the number and shape of testicular follicles, the shape of the seminal vesicle, and the size and color of the accessory gland. The testicular follicle consists of the epithelium lying on a thick basal lamina surrounded by a connective sheath, and a wide lumen filled with germ cells. Large numbers of vesicles and granular materials are observed in the epithelial cell. The two seminal vesicles close-set under a common tunica are composed of the basal lamina, the thick muscular-connective sheath, a mono-layered epithelium, and the vesicular lumen. The vesicles and granules are extensive in the cytoplasm of the flattened epithelial cell, whose apical surface usually has numerous microvilli. The accessory gland is made up of thin muscular-connective sheath, the basal lamina, the epithelium, and the tubular lumen. The epithelial cell with secretory granules has abundant rough endoplasmic reticulum and small vesicles, and its cytoplasm abounds with arrays of three types of granules. Spermiogenesis and sperm ultrastructure of K. paulula was also observed, whose sperm can be distinguished from those of other Cicadellidae by the accessory bodies and mitochondrial derivatives with gap region.

Normal Anatomy, Histology, and Spontaneous Pathology of the Nasal Cavity of the Cynomolgus Monkey (Macaca fascicularis).

The evaluation of inhalation studies in monkeys is often hampered by the scarcity of published information on the relevant nasal anatomy and pathology. We examined nasal cavities of 114 control cynomolgus monkeys from 11 inhalation studies evaluated 2008 to 2013, in order to characterize and document the anatomic features and spontaneous pathology. Compared to other laboratory animals, the cynomolgus monkey has a relatively simple nose with 2 unbranched, dorsoventrally stacked turbinates, large maxillary sinuses, and a nasal septum that continues into the nasopharynx. The vomeronasal organ is absent, but nasopalatine ducts are present. Microscopically, the nasal epithelium is thicker than that in rodents, and the respiratory (RE) and transitional epithelium (TE) rest on a thick basal lamina. Generally, squamous epithelia and TE line the vestibule, RE, the main chamber and nasopharynx, olfactory epithelium, a small caudodorsal region, while TE is observed intermittently along the passages. Relatively high incidences of spontaneous pathology findings, some resembling induced lesions, were observed and included inflammation, luminal exudate, scabs, squamous and respiratory metaplasia or hyperplasia, mucous cell hyperplasia/metaplasia, and olfactory degeneration. Regions of epithelial transition were the most affected. This information is considered helpful in the histopathology evaluation and interpretation of inhalation studies in monkeys.

Bronchial involvement in advanced stage lymphangioleiomyomatosis: histopathologic and molecular analyses

Lymphangioleiomyomatosis (LAM), a rare progressive disease that almost exclusively affects women, is characterized by pulmonary cysts and neoplastic proliferation of smooth muscle-like cells (LAM cells). Airflow obstruction is a physiologic consequence that is commonly observed in LAM and has been attributed to narrowing of peripheral airways. However, histopathologic examinations of the entire airway have been precluded by the limited availability of such specimens. Here, we used explanted lung tissues from 30 LAM patients for a thorough histologic analysis with a special emphasis on the bronchi. We found bronchial involvement by LAM cells and lymphatics in all patients examined. Furthermore, a moderate to severe degree of chronic inflammation (73%), goblet cell hyperplasia (97%), squamous cell metaplasia (83%) of the epithelium, and thickening of basal lamina (93%) were identified in the bronchi. Because LAM cells are transformed by the functional loss of the TSC genes leading to a hyperactivated mammalian target of rapamycin complex 1 (mTORC1) signaling pathway, we confirmed the expression of phospho-p70S6K, phospho-S6, phospho-4E-BP1, and vascular endothelial growth factor (VEGF)-D in LAM cells from all of the patients examined. In contrast, no protein expression of hypoxia-inducible factor 1α, a downstream molecule indicative of mTORC1 activation and leading to VEGF production, was detected in any patient. Our study indicates that late-stage LAM patients commonly have bronchi involved by the proliferation of both LAM cells and lymphatics and that chronic inflammation complicated their disease. Furthermore, the up-regulation of hypoxia-inducible factor 1α, a common event in mTORC1-driven tumor cells, does not occur in LAM cells and plays no role in VEGF-D expression in LAM cells.

AGE-modified basement membrane cooperates with Endo180 to promote epithelial cell invasiveness and decrease prostate cancer survival.

Biomechanical strain imposed by age-related thickening of the basal lamina and augmented tissue stiffness in the prostate gland coincides with increased cancer risk. Here we hypothesized that the structural alterations in the basal lamina associated with age can induce mechanotransduction pathways in prostate epithelial cells (PECs) to promote invasiveness and cancer progression. To demonstrate this, we developed a 3D model of PEC acini in which thickening and stiffening of basal lamina matrix was induced by advanced glycation end-product (AGE)-dependent non-enzymatic crosslinking of its major components, collagen IV and laminin. We used this model to demonstrate that antibody targeted blockade of CTLD2, the second of eight C-type lectin-like domains in Endo180 (CD280, CLEC13E, KIAA0709, MRC2, TEM9, uPARAP) that can recognize glycosylated collagens, reversed actinomyosin-based contractility [myosin-light chain-2 (MLC2) phosphorylation], loss of cell polarity, loss of cell-cell junctions, luminal infiltration and basal invasion induced by AGE-modified basal lamina matrix in PEC acini. Our in vitro results were concordant with luminal occlusion of acini in the prostate glands of adult Endo180(Δ) (Ex2-6/) (Δ) (Ex2-6) mice, with constitutively exposed CTLD2 and decreased survival of men with early (non-invasive) prostate cancer with high epithelial Endo180 expression and levels of AGE. These findings indicate that AGE-dependent modification of the basal lamina induces invasive behaviour in non-transformed PECs via a molecular mechanism linked to cancer progression. This study provides a rationale for targeting CTLD2 in Endo180 in prostate cancer and other pathologies in which increased basal lamina thickness and tissue stiffness are driving factors. © 2014 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

Surface topography and ultrastructural architecture of the tegument of adult Carmyerius spatiosus Brandes, 1898

Adult Carmyerius spatiosus or stomach fluke has an elongate, cylindrical-shaped, straight to slightly curved body, with conical anterior end and truncated posterior end. The worm measures about 8.7–11.2mm in body length and 2.3–3.0mm in body width across the mid-section. When observed by SEM, the tegumental surface in all part of the body appears highly corrugated with ridges and furrows, and having no spines. The ventral surface has more complex corrugation than those of the dorsal surface. Both anterior and posterior suckers have thick edges covered with transverse folds and appear spineless. The genital pore is located at the anterior part of the body. There are two types of sensory papillae on the surface: type 1 is bulbous in shape with nipple-like tips; type 2 has a similar shape with short cilia on the tip. The dorsal surface exhibits similar surface features, but papillae appear less numerous and are smaller. When observed by TEM, the tegument is divided into four layers. The first layer includes the ridges and furrows which are covered by a trilaminate membrane underlined by a dense lamina and coated externally with the glycocalyx. The second layer of the tegument is a narrow region of cytoplasm that contains high concentrations of ovoid electron lucent tegumental granules (TG1), and disc-shaped electron dense tegumental granules (TG2) as well as lysosomes. TG1 close to the surface invariably exocytose their content into bottoms of the ridges, while some TG2 are fused and have their membrane joined up with the surface membrane. The third layer is the widest middle area of the tegument which contains numerous and evenly distributed mitochondria. Both TG1 and TG2 granules are present but in much fewer number than in the first and second layers. The fourth layer is the innermost zone that rests on and couples with a thick basal lamina. The cytoplasm in this layer is loosely packed and contains numerous infoldings of the basal plasma membrane with closely associated mitochondria. It also contains fairly large numbers of TG1 and TG2 granules which are produced and transported to the tegument by one type of tegumental cells lying in rows underneath the muscular layers.

Ischemia/Reperfusion-induced neovascularization in the cerebral cortex of the ovine fetus.

Information on the effects of injury on neovascularization in the immature brain is limited. We investigated the effects of ischemia on cerebral cortex neovascularization after the exposure of fetuses to 30 minutes of cerebral ischemia followed by 48 hours of reperfusion (I/R-48), 30 minutes of cerebral ischemia followed by 72 hours of reperfusion (I/R-72), or sham control treatment (Non-I/R). Immunohistochemical and morphometric analyses of cerebral cortex sections included immunostaining for glial fibrillary acidic protein and collagen type IV (a molecular component of the vascular basal lamina) to determine the glial vascular network in fetal brains and Ki67 as a proliferation marker. Cerebral cortices from I/R-48 and I/R-72 fetuses exhibited general responses to ischemia, including reactive astrocyte morphology, which was not observed in Non-I/R fetuses. Cell bodies of reactive proliferating astrocytes, along with large end-feet, surrounded the walls of cerebral cortex microvessels in addition to the thick collagen type IV-enriched basal lamina. Morphometric analysis of the Non-I/R group with the I/R-48 and I/R-72 groups revealed increased collagen type IV density in I/R-72 cerebral cortex microvessels (p < 0.01), which also frequently displayed a sprouting appearance characterized by growing tip cells and activated pericytes. Increases in cerebral cortex basic fibroblast growth factor were associated with neovascularization. We conclude that increased neovascularization in fetal cerebral cortices occurs within 72 hours of ischemia.

Structure and Barrier Properties of Human Embryonic Stem Cell–Derived Retinal Pigment Epithelial Cells Are Affected by Extracellular Matrix Protein Coating

Extracellular matrix (ECM) interactions play a vital role in cell morphology, migration, proliferation, and differentiation of cells. We investigated the role of ECM proteins on the structure and function of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells during their differentiation and maturation from hESCs into RPE cells in adherent differentiation cultures on several human ECM proteins found in native human Bruch's membrane, namely, collagen I, collagen IV, laminin, fibronectin, and vitronectin, as well as on commercial substrates of xeno-free CELLstart™ and Matrigel™. Cell pigmentation, expression of RPE-specific proteins, fine structure, as well as the production of basal lamina by hESC-RPE on different protein coatings were evaluated after 140 days of differentiation. The integrity of hESC-RPE epithelium and barrier properties on different coatings were investigated by measuring transepithelial resistance. All coatings supported the differentiation of hESC-RPE cells as demonstrated by early onset of cell pigmentation and further maturation to RPE monolayers after enrichment. Mature RPE phenotype was verified by RPE-specific gene and protein expression, correct epithelial polarization, and phagocytic activity. Significant differences were found in the degree of RPE cell pigmentation and tightness of epithelial barrier between different coatings. Further, the thickness of self-assembled basal lamina and secretion of the key ECM proteins found in the basement membrane of the native RPE varied between hESC-RPE cultured on compared protein coatings. In conclusion, this study shows that the cell culture substrate has a major effect on the structure and basal lamina production during the differentiation and maturation of hESC-RPE potentially influencing the success of cell integrations and survival after cell transplantation.