Disruption Of Basal Lamina Research Articles

Cutaneous Superficial Basal Cell Carcinomais a Basal Cell Carcinoma In Situ: Electron Microscopy of a Case Series of Basal Cell Carcinomas.

Basal cell carcinoma (BCC) is the most common skin cancer. Skin cancers may present either as a non-invasive tumor or an invasive malignancy. The terminology of carcinoma in situ is used when the tumor is either just limited to epidermis or not present as single cells or nests in the dermis. However, currently the terminology superficial BCC is inappropriately used instead of BCC in situ when the skin cancer is limited to epidermis. In this study we compare the pathologic changes of superficial, nodular, and infiltrative BCCs using electron microscopy to identify the ultrastructural characteristics and validate the previously proposed terminology. Three cases of BCC (superficial BCC, nodular BCC, and infiltrative BCC) diagnosed by dermatopathologists at our institute were selected for review. Paraffin block tissues from these cases were sent for electron microscopy studies which demonstrated disruption of basal lamina in both nodular and infiltrative type of BCC, while it remains intact in BCC superficial type after extensive examination. Therefore, similar to other in situ skin cancers, there is no invasion of the neoplasm in superficial BCC into the dermis. Hence, the older term superficial BCC should be appropriately replaced with the newer terminology BCC in situ.

Consequences of Microwave oven Exposed Diet on The Basal Lamina of Seminiferous Tubules of Mice

Usage of electronic gadgets like microwave oven is increasing day by day that heats the food by exposing it to electromagnetic radiations which has many hazardous effects on human health including fertility. Aim: To find the effects of microwave oven exposed diet on basal lamina of seminiferous tubules of mice alongwith protective effects of Mentha piperita and melatonin on the same tissue. Study Design: Randomized control trial. Methodology: Adult male mice (n=32) were divided into four groups. Control group (G1) received standard pellets prepared for mice. Second group (G2) was given mice pellets exposed to microwave oven. Third group (G3) received Mentha Piperita leaf extract along with mice pellets exposed to microwave oven and the fourth group (G4) received oral melatonin along with pellets exposed to microwave oven. Later their testicular tissue was removed for histological examination while basal lamina disruption was assessed by scoring. Data analyzed by SPSS 22.0v. Results: In group G2, there was slight disruption in the basal lamina in 75% of the cases while in experimental group G3, there was slight disruption of basal lamina only in 12.5% of the cases. However, in group G4, only 25% specimen had slight disruption of basal lamina Conclusion: It was concluded that microwave oven exposed diet produced severe disruption of basal lamina in group G2 that decreased in Mentha piperita and melatonin treated groups. However, Mentha piperita treated group produced better results than melatonin treated group. Keywords: Mice, Testis, Basal Lamina, Mentha piperita and Melatonin

A brief insight on the etiopathogenesis of oral lichen planus

Oral Lichen Planus (OLP) is a relatively common, systemic, immune based disease. It usually, involves the oral mucosa, but other sites like the skin, genital mucosa, and scalp are also affected. In this auto-cytotoxic muco-cutaneous disease, the T lymphocytes trigger apoptosis of epithelial cells leading to chronic inflammation. OLP is mediated by both antigen specific and nonspecific pathways. Antigen presentation through basal keratinocytes and antigen-specific keratinocyte destruction by CD8+ cytotoxic T-cells are within the specific pathway spectrum. Whilst the nonspecific methods involve mast cell degeneration; leading to granular release and matrix metalloproteinase (MMP) activation. The process ultimately results in clustering of T cells in the upper layer of lamina propria, basal lamina disruption, T-cell inflow within the epithelium, along with apoptotic death of keratinocytes. The normal oral mucosa may be an immune privileged site and breach of the immune barrier could result in OLP and possibly other autoimmune oral mucosal diseases owing to deficient antigen-specific TGF-β1-mediated immunosuppression. However, many among these immuo pathologic mechanisms are still shroud in mystery.

EFFECTS OF CANNABIS SATIVA EXTRACT AND ITS COMBINATION WITH VITAMIN-C ADMINISTRATION ON HISTOMORPHOLOGICAL CHANGES IN THE TESTICULAR TISSUE OF MALE MICE

OBJECTIVES: To observe morphological changes in testicular tissue of adult male BALB/c mice after administration of Cannabis sativa extract and to investigate the protective role of vitamin-C against cannabis-induced histopathological changes. METHODS: Eighteen BALB/c mice (31-40g) were divided into 3 groups. Group-I was control group, Group-II received oral cannabis extract (14mg/kg) and Group-III received cannabis extract (14mg/kg) orally along with vitamin-C (100 mg/kg) for 45 days. At the end of experiment, variations in body weight, testicular weight and histological changes in testicular tissues of mice were observed. RESULTS: Average increase in weight was 7.000±0.77gm, 4.833±0.87gm & 5.833±0.79gm in group-I, group-II and group-III respectively. Weight reduction in right and left testis was 42.30±0.465mg and 42.45±0.424mg in group-I, 38.85±0.523 mg and 39.35±0.520 mg in group-II, and 42.38±0.465 mg and 42.23±0.424 mg in group-III animals respectively (p<0.01). Mean seminiferours epithelium thickness was 4.367±0.302 µm in group-II, 5.683±0.104 µm and 6.417±0.116 µm in Group-I and group-III respectively (p<0.001). Average germ cells count (cells/tubule) in group-II was reduced (256.25±6.58) as compared to group-I (373.50±4.063) and group-III (366.25±3.442). Histological changes observed after the treatment were height of germinal epithelium, basal lamina disruption, seminiferous tubules shrinkage, scanty cytoplasm with shrunken nuclei and germinal epithelium detachment from basal lamina in group-II animals as compared to group-I and group-III. CONCLUSION: The cannabis administration significantly declines the weight, size of testes, germ cells count and the height of seminiferours epithelium in animals. Vitamin-C had a protective role on cannabis induced toxic changes in the testes of the mice.

An Anthelmintic Assessment of Balanites aegyptiaca Fruits on Some Multiple Drug Resistant Gastrointestinal Helminthes Affecting Sheep

The goal of this study is to evaluate the effect of Balanites aegyptiaca methanolic extract fruits as an alternative sustainable source of anthelmintic drugs, on adult gastrointestinal multiple drug resistant helminthes and clarify how B. aegyptiaca fruits directly affect helminthes. To fulfill this study, three hundred and fifty sheep reared in a private farm, in Giza Governorate, had been subjected to clinical investigation. Of them, fifty five severely affected sheep had been isolated relying on their clinical history of multiple anthelmintic resistances, to be slaughtered. The post mortem examination revealed that the animals had a mixed infection with Haemonchus contortus, Moniezia expansa and Fasciola gigantica. Light and scanning electron microscopic studies were undertaken to evaluate changes in body wall of these adult worms following 24 h exposure to methanolic extract of B. aegyptiaca fruits utilizing concentrations of 10, 50, 100, 200 and 400 µg/ml. The main target organ that was highly affected was the body wall, whose damage was observed by light and scanning electron microscopic studies. These alterations occurred in particular sequences in response to Balanites extract concentration, initiated with swelling that was even disrupted, resulting in sloughing off cuticle or tegument, and finally exposure and disruption of basal lamina. This damage would induce disturbance for many of the physiological processes associated with the body wall, especially nutrition and immunoprotection. This plant may offer an opportunity for a new natural anthelmintic and an alternative source for the control of such infectious disease in sheep.

Modes of action of squamocin in the anal papillae of Aedes aegypti larvae

The morphological and the primary transcript levels of the genes for aquaporin AaAQP4 and the membrane transporter AaV-H+-ATPase in the anal papillae of A. aegypti larvae exposed to squamocin were evaluated. Squamocin caused great vacuolization in the anal papillae wall, basal lamina disruption with loss of canalicular spaces, and disorganization of the cuticular layers. There was also decrease of AaAQP4 transcript levels and inhibition of AaV-H+-ATPase in the anal papillae of larvae. Morphological together with decrease in the AaAQP4 and AaV-H+-ATPase primary transcript levels suggest that squamocin may affect osmoregulation and ion-regulation of this insect followed by death.

Copper Nanoparticles Differentially Target Testis of the Catfish, Clarias batrachus: In vivo and In vitro Study

Present study examined the impact of copper (Cu) as nanoparticle (Cu-NPs) and soluble form (CuSO4) in adult male catfish Clarias batrachus. Following treatment, upregulation of transcripts of several testis-related genes and increased androgen levels were evident. Though profound promotional effect was seen at gene and androgen levels, structural analysis revealed completely different scenario. Histological observations of testis showed disruption of basal lamina, distinct spermatogonium and enlarged spermatocytes in the treated groups. Quite alarmingly, transmission electron microscopic analysis showed damage of testicular lumen with irregular basal lamina and distinctive spermatocytes. In addition, Sertoli cells and erythrocytes exhibited damage after Cu treatments. Cytotoxic effects of Cu in catfish primary testis and TM3 Leydig cells showed cytotoxicity which is correlated well with morphological and flow cytometric analysis. Taken together, the results demonstrated that low dose of Cu/Cu-NPs can be detrimental to catfish testicular recrudescence vis-a-vis reproduction and the toxic effect manifest drastically in in vitro cultures.

In vitro tegumental alterations on adult Fasciola gigantica caused by mefloquine.

Emergence of drug-resistant Fasciola strains has drawn the attention of many authors to alternative drugs. The purpose of this study is to explore the in vitro effect of the antimalarial mefloquine against adult Fasciola gigantica. Light and scanning electron microscopic observations could be used to determine the target of the drug following 6 and 12h of incubation in medium containing mefloquine at three different concentrations 10, 20 and 30μg/mL, as morphological changes could be observed. These changes occurred in definite sequences in response to mefloquine, and were consisted of swelling, vacuolization that was later disrupted, leading to desquamation of the tegument, resulting in exposure and disruption of basal lamina and the dislodging of spines. It is concluded that mefloquine presented itself as a drug that might become important in trematode chemotherapy, with the tegument being an important drug target.

Mercuric chloride induced toxicity responses in the olfactory epithelium of Labeo rohita (HAMILTON): a light and electron microscopy study

Bioaccumulation of mercury and histomorphological changes in the olfactory epithelium of Labeo rohita were investigated after exposing the fish to two sublethal concentrations of HgCl₂ (66 and 132 μg/L) for 15 and 30 days. Mercury deposition increased in the tissue significantly (p < 0.05) with dose- and duration-dependent manner. Severe damage to the olfactory epithelium was evident. When fish exposed to 66 μg/L for 15 days, the histology of olfactory epithelium exhibited that mucous cell proliferation was upregulated and cell size was significantly increased from the control. Similar trends were found in 30 days exposure in both treated groups. Histology showed that mercury induced degeneration of columnar sensory cells, supporting cells and ciliated non-sensory cells and induced basal cell proliferation. Basal cell hyperplasia led to form intraepithelial proliferative lesion, thickening of epithelium, basal lamina disruption and cyst formation. Scanning electron microscopy revealed that mercury exposure at 66 μg/L caused clumping and loss of cilia, erosion in microridges on the supporting cells and proliferation of mucous cell opening. Complete degeneration of ciliated cells and cyst formation was observed in the fish when exposed to 132 μg/L HgCl₂. This result suggests that prolonged exposure to mercury might cause irreversible damage to the olfactory epithelium and impair the olfactory function of fish.

Epithelial-Mesenchymal Transition as a Possible Mechanism of Semicircular Canal Morphogenesis in Chick Inner Ear

Semicircular canals are sensory organs for balance, consisting of fluid-filled tubules that are arranged perpendicularly to each other in inner ear. The precise mechanism of the morphogenesis of this unique organ is still under investigation. Semicircular canals arise from the flattened pouch of epithelium. The centers of two apposing wall of the pouch approach each other and form a fusion plate. The clearing of the fusion plate makes a hole and leaves the remaining tissue as semicircular canals. Three mechanisms have been proposed for this clearing: programmed cell death, epithelial-mesenchymal transition, and retraction of the cells in the fusion plate to surrounding semicircular canals. Previous studies have revealed programmed cell death in the fusion plate, although other two hypotheses were not disproved. Here we examined the contribution of epithelial-mesenchymal transition and epithelial retraction to the morphogenesis of semicircular canals. We analyzed immunohistochemically the structural change in the epithelium of the developing fusion plate using molecular markers, basal lamina component laminin, cytoskeletal F-actin, and cellular junctional marker beta-catenin. Our observation revealed that fusion plate epithelium lost its apico-basal polarity and intermingled with facing fusion plate cells, associated with the disruption of basal lamina. Moreover, there were several cells with mesenchymal appearance adjacent to the torn basal lamina. We also found the merging of apposing basal laminae at the border between forming canal and breaking fusion plate. These observations suggest that the epithelial-mesenchymal transition, rather than the epithelial retraction, may be responsible for clearing fusion plate cells.

Cellular basis of verruciform xanthoma: immunohistochemical and ultrastructural characterization

Verruciform xanthoma (VX) holds two basic pathogenic interests: (1) Why and how do macrophage foam cells accumulate exclusively in the sub-basal papillae? and (2) What underlies the disease chronicity? Moreover, an unsolved question is which came first - epithelial hyperplasia or foam cell collection? We analyzed 36 oral mucosal lesions to dissect a series of linked cellular changes in VX using immunohistochemical and ultrastructural techniques. Macrophage scavenger receptor-1 (MSR-1), monocyte chemoattractant protein-1 (MCP-1), CCR2, and oxidized low-density lipoprotein (ox-LDL) were all expressed by foam cells. VX epithelium showed reactivity for MCP-1, HLA-DR and IL8 in varying degrees, and showed a nearly 40% reduction in Langerhans cell density. In sub-epithelial inflammatory infiltrates, CD8+ T cells preponderated (>70%), but only a minority were positive for granzyme B (<1%). Keratinocyte/basal lamina complex exhibited disruption of basal lamina, squamatization and cytolysis of basal cells, fragmentation of desmosomes, and intraepithelial migration of macrophages. In severely inflamed papillae, necrotic foam cells were scavenged by adjacent macrophages. Under synergistic regulation of T cells, MCP-1/CCR2-mediated macrophage recruitment in the sub-basal papillae and the lysosomal engulfment of epithelial lipids by MSR-1-bearing macrophages may be central in VX formation. Once developed, ox-LDL-induced foam cell necrosis and macrophage-dependent debris disposal may cyclically perpetuate VX.

Gelatinase A (MMP-2) Is Necessary and Sufficient for Renal Tubular Cell Epithelial-Mesenchymal Transformation

Progressive renal interstitial fibrosis and tubular atrophy represent the final injury pathway for all commonly encountered forms of renal disease that lead to end-stage renal failure. It has been recently recognized that myofibroblastic cells are the major contributors to the deposition of interstitial collagens. While there are several potential cellular sources of myofibroblasts, attention has focused on the transformation of the organized tubular epithelium to the myofibroblastic phenotype, a process potently driven both in vitro and in vivo by transforming growth factor-beta1 (TGF-beta1). Integrity of the underlying basal lamina provides cellular signals that maintain the epithelial phenotype, and disruption by discrete proteases could potentially initiate the transformation process. We demonstrate that TGF-beta1 coordinately stimulates the synthesis of a specific matrix metalloproteinase, gelatinase A (MMP-2), and its activator protease, MT1-MMP (MMP-14), and that active gelatinase A is absolutely required for epithelial-mesenchymal transformation induced by TGF-beta1. In addition, purified active gelatinase A alone is sufficient to induce epithelial-mesenchymal transformation in the absence of exogenous TGF-beta1. Gelatinase A may also mediate epithelial-mesenchymal transformation in a paracrine manner through the proteolytic generation of active TGF-beta1 peptide. MT1-MMP and gelatinase A were co-localized to sites of active epithelial-mesenchymal transformation and basal lamina disruption in the rat remnant kidney model of progressive renal fibrosis. These studies indicate that a discrete matrix metalloproteinase, gelatinase A, is capable of inducing the complex genetic rearrangements that characterize renal tubular epithelial-mesenchymal transformation.

Temporary Disruption of the Retinal Basal Lamina and Its Effect on Retinal Histogenesis

An experimental paradigm was devised to remove the retinal basal lamina for defined periods of development: the basal lamina was dissolved by injecting collagenase into the vitreous of embryonic chick eyes, and its regeneration was induced by a chase with mouse laminin-1 and α2-macroglobulin. The laminin-1 was essential in reconstituting a new basal lamina and could not be replaced by laminin-2 or collagen IV, whereas the macroglobulin served as a collagenase inhibitor that did not directly contribute to basal lamina regeneration. The regeneration occurred within 6 h after the laminin-1 chase by forming a morphologically complete basal lamina that included all known basal lamina proteins from chick embryos, such as laminin-1, nidogen-1, collagens IV and XVIII, perlecan, and agrin. The temporary absence of the basal lamina had dramatic effects on retinal histogenesis, such as an irreversible retraction of the endfeet of the neuroepithelial cells from the vitreal surface of the retina, the formation of a disorganized ganglion cell layer with an increase in ganglion cells by 30%, and the appearance of multiple retinal ectopias. Finally, basal lamina regeneration was associated with aberrant axons failing to correctly enter the optic nerve. The present data demonstrate that a transient disruption of the basal lamina leads to dramatic and probably irreversible aberrations in the histogenesis in the developing central nervous system.

Disruption of the retinal basal lamina during early embryonic development leads to a retraction of vitreal end feet, an increased number of ganglion cells, and aberrant axonal outgrowth

Bacterial collagenase was injected into the vitreous of the eye of chick and quail embryos. Immunocytochemical and ultrastructural studies revealed that the collagenase dissolved the retinal basal lamina of the injected eye. The basal lamina disruption was first detectable 1 hour after enzyme injection and was complete within 3 hours. With further development, the retinal basal lamina was not reestablished; newly developing neuroepithelium in the peripheral retina, however, generated an intact basal lamina. Western blot analysis showed that Clostridial collagenase degraded various collagens but spared noncollagenous proteins. Basal lamina disruption of embryonic day 3 to 6 retinae led to the retraction of the end feet of the neuroepithelial cells, caused an increase in the number of Islet-1+ cells (most likely ganglion cells), an increase in the thickness of the optic fiber layer, and aberrant growth of optic axons on their way toward the optic disc. None of these changes were observed when retinal basal laminae were disrupted at later stages of development. The present data demonstrate that the retinal basal lamina, by anchoring the neuroepithelial cells to the pial surface of the retina, has an important function in the development of the normal cytoarchitecture of this structure. It is proposed that the altered extracellular environment in the vitreal part of the retina, resulting in the retraction of the neuroepithelial end feet, is responsible for the increased number of Islet-1+ cells and the aberrant axonal navigation.

Phosgene effects on F-actin organization and concentration in cells cultured from sheep and rat lung

Pulmonary edema and immunosuppression of the lung are primary causes of debilitation and death from phosgene gas exposure. The pathophysiology that gives rise to these conditions shares a common clinical pathway. However, the target cells and lesions that disrupt normal barrier function and immune response of the lung are complex and poorly understood. Using confocal laser microscopy and FITC-conjugated phalloidin, we have studied the effects of phosgene on F-actin in endothelial cells from sheep pulmonary arteries and epithelial cells from rat tracheal explants. Image analyses from attached culture systems indicate that F-actin was a sensitive target molecule in both species. Exposures ranging from 0.15 to 1.0 x LCt50 for sheep in vivo (3300 ppm.min) produced immediate, dose-dependent decreases in average F-actin content of cultured endothelial cells. Dense peripheral bands and stress fibers were diminished and partially disrupted but were not destroyed by these doses. Changes in ultrastructure and the permeability barrier of endothelial tissues included separation of basal lamina and development of paracellular leakage paths. Phosgene also decreased the F-actin in airway epithelial cells and potentiated phenotypic transformations that gave rise to progeny with dendritic processes. Differences in endothelial and airway epithelial response indicate that the cytoskeletal effects of phosgene were cell-type specific. Disruption of basal lamina, depletion of F-actin, and development of endothelial leakage paths may contribute to decreased barrier function and increased permeability of vascular tissues. Phosgene-induced transformations that involved F-actin reorganization and appearance of dendritic cells among airway epithelial may affect other functions of the lung.

Bacterial collagenase disrupts extracellular matrix and opens blood-brain barrier in rat

Bacterial collagenase causes hemorrhagic necrosis of brain. We studied the enzyme's effect on blood-brain barrier (BBB) permeability and extracellular matrix (ECM) structure by radiolabeled tracers and electron microscopy. Adult rats had intracerebral injection of bacterial collagenase. Brain uptake from blood of [14C]sucrose was measured in 24 rats 0.5 h to 14 days after injection. 12 rats had ultrastructural studies 1 h after collagenase injection. Brain uptake of [14C]sucrose is maximally increased at 0.5 h, remaining significantly increased for 7 days. Ultrastructurally, some vessels had widening of basal lamina while others had severe disruption of basal lamina with stretching of endothelial cells. We conclude that bacterial collagenase disrupts ECM and opens BBB.

Age-dependent disruption of basal lamina and extracellular matrix formation in l-proline analog treated otic explants

l-azetidine-2-carboxylic acid, LACA, a naturally occurring vegetable imino acid, can be incorporated into mammal proteins instead of proline. This incorporation has an especially inhibitive effect on collagen secretion. Exposure of embryonic mouse inner ear explants to LACA causes dysmorphogenesis and retarded differentiation, reduces the number of collagen fibrils in the perilymphatic spaces and capsules, and gives rise to a dose-dependent derangement of the basal lamina. In control specimens, both in vivo and in vitro, the inner ear epithelia had a dense contiguous basal lamina overlying a well-developed network of collagen fibrils. When the inner ears were exposed to LACA at a concentration of 150 μg per ml of medium, there was a loss of the collagen network and gaps appeared in the basal lamina. At exposure to 300 μg LACA/ml, scarcely any collagen fibrils were present and the basal lamina was disrupted in many areas, especially beneath the sensory epithelia.

Ultrastructural distribution of sulfated glycosaminoglycans in epithelial-mesenchymal interface of developing rat tooth germs.

We investigated the ultrastructural distribution of sulfated glycosaminoglycans in the epithelial-mesenchymal interface of tooth germs by use of the high-iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining and enzymatic digestion method. At an early stage in odontoblast differentiation, HID-TCH-SP stain deposits were sparsely distributed in the basement membrane and in the intercellular spaces. Subsequently, as formation of the initial predentin matrix began, HID-TCH-SP stain deposits were densely distributed in the interfibrillar spaces and the basement membrane. Testicular hyaluronidase digested most of those in the progenitor pre-dentin, whereas those in the region of basal lamina resisted enzymatic digestion. Testicular hyaluronidase-resistant HID-TCH-SP stain deposits were susceptible to heparitinase, indicating that the sulfated glycosaminoglycan in the basal lamina is heparan sulfate. Furthermore, the heparan sulfate tended to be regularly arranged at the sites of internal and external lamina densa. However, as progenitor pre-dentin matrix formation proceeded, the numbers of stain deposits temporarily increased and their distribution pattern became irregular, finally tending to disappear with the disruption of basal lamina.

Stromal changes in early invasive and non-invasive breast carcinoma: an ultrastructural study.

Six examples of histologically diagnosed, non-invasive breast carcinomas were studied by electron microscopy to elucidate the ultrastructural features for an accurate diagnosis of in situ carcinoma. The results obtained revealed two patterns of basal lamina/stromal cells relationship. One pattern showed intact basal lamina with associated periductal stromal cells consisting entirely of fibroblasts, the other pattern showed disruption of basal lamina by gaps and malignant cell protrusions with associated stromal cells consisting of both fibroblasts and myofibroblasts. As myofibroblasts are not a component of normal breast stroma but are known to be a prominent feature in the stroma of infiltrating breast carcinoma, the present observations suggest that myofibroblastic proliferation around in situ carcinoma represents an early sign of carcinomatous infiltration. Hence the definitive diagnosis of non-invasive carcinoma of the breast requires an intact basal lamina and a complete absence of a myofibroblastic reaction.

Elvax 40P implants: Sustained, local release of bioactive molecules influencing mammary ductal development

Elvax 40P (EVX), an ethylene vinyl-acetate copolymer, has been well characterized as an implant material that causes no inflammatory response and is capable of the sustained, local release of a wide variety of undenatured macromolecules in vivo. To investigate the usefulness of this material in developmental studies we examined the effect of EVX implants containing either deoxycorticosterone acetate (DCA) or testicular hyaluronidase on alveolar differentiation and ductal growth in the mouse mammary gland. DCA implants produced localized alveolar differentiation on ducts, while implants containing Thase caused basal lamina disruption at the duct's growing tip, resulting in epithelial dysplasias. We conclude that EVX implants allow assessment of the primary (nonsystemic) effects of biologically active molecules on developing tissue and should therefore have a variety of interesting experimental uses.