Degeneration Of Epithelial Cells Research Articles

Protective Effect of L- Carnitine Against Formaldehyde-Induced Kidney, Liver and Testicular Damage in Rabbits, A Histopathological Study

Formaldehyde (dissolved in market milk) was given every day by gavage to male rabbits for 90 days and the histological changes in the liver, kidney and testicles were examined using paraffin method. This compound has caused hepatotoxicity which included portal inflammation and infiltration by mononuclear inflammatory cells of the hepatocytes. This compound has also caused nephrotoxicity which included tubular dilatation and hydropic degeneration of tubular epithelial cells. The effect on the testicles included atrophy of some seminiferous tubules with shedding of disintegrated epithelial cells into the lumina as well as decrease in the number of the sperms. When given to the formalin exposed rabbits, L-Carnitine caused recovery of the changes in the liver, kidneys and testicles i.e. it has been succeeded in playing a protective role against formalin hepatotoxicity, nephrotoxicity and testicular damage.

Disruption of retinal pigment epithelial cell properties under the exposure of cotinine

Cigarette smoking is a major risk factor for age-related macular degeneration (AMD), in which progressive retinal pigment epithelial (RPE) cell degeneration is a major pathological change. Nicotine is a major biologically active component in cigarette smoke. It is continuously catabolized into cotinine, which has longer half-life and higher concentration in tissue cells and fluids. Here we hypothesized that continuous exposure of cotinine has more potent effects on human RPE cell properties than nicotine. Human RPE cell line (ARPE-19) was treated continuously with 1–2 µM of nicotine and/or cotinine for 7 days. RPE cells treated with 2 μM cotinine and nicotine-cotinine mixture has lower MTT signals without significant changes in cell apoptosis or integrity. Moreover, RPE cell migration was retarded under cotinine treatments, but not nicotine. Both nicotine and cotinine treatments attenuated the phagocytotic activity of RPE cells. In addition, cotinine and nicotine-cotinine mixture suppressed VEGF and IL-8 expression and upregulated TIMP-2 expression. Expressions of autophagy genes were upregulated by the cotinine treatment, whereas expressions of epithelial-to-mesenchymal transition markers were downregulated. In conclusion, our study, for the first time, demonstrated that cotinine, rather than nicotine, affects the properties of RPE cells in vitro, which could explain the smoking-induced RPE pathology.

Ectopic germinal center and megalin defect in primary Sjogren syndrome with renal Fanconi syndrome

BackgroundThis study reports the clinical and pathological features of 12 cases of primary Sjogren syndrome (pSS) with renal involvement presenting with proximal tubular dysfunction in a single center, and investigates the possible correlation of ectopic germinal center formation and megalin/cubilin down-expression.MethodClinical and pathological records were reviewed. Immunohistochemistry was carried out to detect megalin, cubilin, CD21 and IL-17 expression.ResultsPatients presented with different degrees of proximal renal tubule lesion and decreased estimated glomerular filtration rate (eGFR). Renal biopsy revealed tubulointerstitial nephritis, with tubular epithelial cell degeneration, tubular atrophy, interstitial inflammation and focal fibrosis. Immunohistochemistry revealed decreased expression of megalin and cubilin, two important multiligand protein receptors on the brush border of proximal tubular epithelial cells. IL-17 secreted by Th17 subtype effector T cells was diffusely detected in the renal proximal tubule, with a negative correlation of IL-17 and megalin expression. In addition, ectopic germinal centers characterized by CD21+ follicular dendritic cells were present in the renal interstitium. In patients with a decreased eGFR, treatment with 4 weeks of glucocorticoid therapy resulted in an improved eGFR in 75% of patients.ConclusionWe report 12 cases of pSS characterized by Fanconi syndrome. The decreased megalin and cubilin expression may contribute to the proximal tubular reabsorption defect, possibly secondary to Th17 infiltration and formation of ectopic germinal centers.

Antiestrogenic Activity of Triptolide in Human Breast Cancer Cells MCF-7 and Immature Female Mouse.

Preclinical Research To investigate the antiestrogenic activity of triptolide in human breast cancer cell line MCF-7 and immature female C57BL/6 mouse. The effects of triptolide on cell proliferation, cell cycle, and the expression of estrogen receptor alpha (ERα) and progesterone receptor (PR) were examined in MCF-7 cells. In vivo antiestrogenic effects of triptolide were observed after cotreatment of mice with E2 and triptolide for 4 days. Triptolide dose- and time-dependently inhibited cell growth in untreated or E2 -treated MCF-7 cells, which was associated with increased S phase arrest. Furthermore, triptolide down regulated the expression of ERα and PR in cells. The expression of ERα and PR in combined group of triptolide with E2 was much higher than that of triptolide alone. Triptolide decreased the E2 -induced uterine weight in mice, while triptolide alone had no effect. Triptolide treatment (90 μg/kg) resulted in extensive degeneration and necrosis of uterine epithelial cells, whereas the same concentration of triptolide in combination with E2 caused morphologic changes in epithelial cells from simple columnar to ellipse, without destruction. Triptolide showed antiestrogenic activity in vitro and in vivo, and the down regulation of ERα and PR expression may be its underlying mechanisms. Drug Dev Res 78 : 164-169, 2017. © 2017 Wiley Periodicals, Inc.

Autophagy Regulates Proteasome Inhibitor-Induced Pigmentation in Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells.

The impairment of autophagic and proteasomal cleansing together with changes in pigmentation has been documented in retinal pigment epithelial (RPE) cell degeneration. However, the function and co-operation of these mechanisms in melanosome-containing RPE cells is still unclear. We show that inhibition of proteasomal degradation with MG-132 or autophagy with bafilomycin A1 increased the accumulation of premelanosomes and autophagic structures in human embryonic stem cell (hESC)-derived RPE cells. Consequently, upregulation of the autophagy marker p62 (also known as sequestosome-1, SQSTM1) was confirmed in Western blot and perinuclear staining. Interestingly, cells treated with the adenosine monophosphatedependent protein kinase activator, AICAR (5-Aminoimidazole-4-carboxamide ribonucleotide), decreased the proteasome inhibitor-induced accumulation of premelanosomes, increased the amount of autophagosomes and eradicated the protein expression of p62 and LC3 (microtubule-associated protein 1A/1B-light chain 3). These results revealed that autophagic machinery is functional in hESC-RPE cells and may regulate cellular pigmentation with proteasomes.

Antimicrobial agents, triclosan, chloroxylenol, methylisothiazolinone and borax, used in cleaning had genotoxic and histopathologic effects on rainbow trout

Triclosan (TRC), chloroxylenol (PCMX) and methylisothiazolinone (MIT) have been commonly used as an antimicrobial in soaps while borax (BRX) is used in household cleaning. After using these chemicals, they are washed down drains and getting into the aquatic ecosystem in which they may affect aquatic living organisms. In the present study, the chronic effects of TRC, PCMX, MIT and BRX on genotoxicity, gene expression and histopathology of rainbow trout (Oncorhynchus mykiss) were evaluated for 40 days under semi static condition. The comet assay results indicated that MIT, TRC and PCMX caused significant DNA damage to erythrocytes of the fish. Transcription of SOD, GPX1, GPX2, GSTA, HSP90BB, HSP90BA, CAT, and HSC70A genes were significantly regulated as a result of TRC, PCMX, MIT, and BRX exposure except PCMX exposed GSTA gene. Histological lesions were detected in gills, spleen liver, and trunk kidney of the fish. Lamellar fusion, hyperplasia and epithelial necrosis in gills, melanomacrophage centers and splenic necrosis in spleen, pyknotic nucleus, fat vacuoles, necrotic hepatocytes in liver, cloudy swelling in the tubules, renal tubule epithelial cells degeneration, glomerular capillaries dilation and glomerulus degeneration in kidney, were observed. Our study demonstrates the chronic toxic effect of TRC, PCMX, MIT, and BRX is high in rainbow trout. Therefore, we should be more careful when using these chemicals for cleaning in order to protect aquatic environment.

The effect of starvation and re-feeding on mitochondrial potential in the midgut of Neocaridina davidi (Crustacea, Malacostraca).

The midgut in the freshwater shrimp Neocaridina davidi (previously named N. heteropoda) (Crustacea, Malacostraca) is composed of a tube-shaped intestine and a large hepatopancreas that is formed by numerous blind-ended tubules. The precise structure and ultrastructure of these regions were presented in our previous papers, while here we focused on the ultrastructural changes that occurred in the midgut epithelial cells (D-cells in the intestine, B- and F- cells in the hepatopancreas) after long-term starvation and re-feeding. We used transmission electron microscopy, light and confocal microscopes and flow cytometry to describe all of the changes that occurred due to the stressor with special emphasis on mitochondrial alterations. A quantitative assessment of cells with depolarized mitochondria helped us to establish whether there is a relationship between starvation, re-feeding and the inactivation/activation of mitochondria. The results of our studies showed that in the freshwater shrimp N. davidi that were analyzed, long-term starvation activates the degeneration of epithelial cells at the ultrastructural level and causes an increase of cells with depolarized (non-active) mitochondria. The process of re-feeding leads to the gradual regeneration of the cytoplasm of the midgut epithelial cells; however, these changes were observed at the ultrastructural level. Additionally, re-feeding causes the regeneration of mitochondrial ultrastructure. Therefore, we can state that the increase in the number of cells with polarized mitochondria occurs slowly and does not depend on ultrastructural alterations.

Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids.

Neovascular age-related macular degeneration (nvAMD) is characterized by choroidal blood vessels growing into the subretinal space, leading to retinal pigment epithelial (RPE) cell degeneration and vision loss. Vessel growth results from an imbalance of pro-angiogenic (e.g., vascular endothelial growth factor [VEGF]) and anti-angiogenic factors (e.g., pigment epithelium-derived factor [PEDF]). Current treatment using intravitreal injections of anti-VEGF antibodies improves vision in about 30% of patients but may be accompanied by side effects and non-compliance. To avoid the difficulties posed by frequent intravitreal injections, we have proposed the transplantation of pigment epithelial cells modified to overexpress human PEDF. Stable transgene integration and expression is ensured by the hyperactive Sleeping Beauty transposon system delivered by pFAR4 miniplasmids, which have a backbone free of antibiotic resistance markers. We demonstrated efficient expression of the PEDF gene and an optimized PEDF cDNA sequence in as few as 5 × 103 primary cells. At 3 weeks post-transfection, PEDF secretion was significantly elevated and long-term follow-up indicated a more stable secretion by cells transfected with the optimized PEDF transgene. Analysis of transgene insertion sites in human RPE cells showed an almost random genomic distribution. The results represent an important contribution toward a clinical trial aiming at a non-viral gene therapy of nvAMD.

Cytomorphometric analysis and morphological assessment of oral exfoliated cells in type 2 diabetes mellitus and healthy individuals: A comparative study.

Context:Oral exfoliative cytology is a simple, nonaggressive technique that is well accepted by patients. Therefore, it is an attractive option, which aids in the diagnosis and observation of epithelial atypias associated with oral mucosal diseases.Aims:The aim of this study was to evaluate and compare the quantitative and qualitative alterations in exfoliative smears from type 2 diabetics and healthy individuals.Patients and Methods:The study includes 30 type 2 diabetics and 30 healthy persons of both sexes. PAP and hematoxylin and eosin (H and E) stained smears were prepared from buccal mucosa (BM), tongue (T), floor of the mouth (FOM), and palate (P). Under a light microscope, 50 clearly defined unfolded epithelial cells were quantitatively evaluated for cellular area (CA), nuclear area (NA), and cellular-to-nuclear area ratio (CA:NA) and assessed for morphological features.Statistical Analysis:Collected data was manually entered into the Statistical Package for the Social Sciences version 13.5 for analysis. Student's t-test was used at 95% confidence interval.Results:Quantitative assessment of the overall mean CA was less, mean NA was more, and mean CA:NA was less in diabetics than that in healthy persons at all the four sites. Diabetic oral cells showed qualiative cytoplasmic and nuclear alterations: cytoplasmic vacuoles, karyorrhexis, karyolysis, pyknosis, peri-nuclear halo, binucleation, nuclear vacuoles, inflammation, and microbial colonies.Conclusion:Oral cytology from type 2 diabetics is associated with detectable cytomorphological changes with alteration in size of the cell and nucleus, which is site specific, indicating epithelial cell degeneration in cytoplasm and nucleus.

Experimental poisoning by Niedenzuella stannea in cattle and corresponding detection of monofluoroacetate

ABSTRACT: In Brazil sodium monofluoroacetate (MFA) containing plants are represented by 16 species that belong to three principle genera: Palicourea ( Rubiaceae ), Amorimia ( Malpighiaceae ), and Tanaecium ( Bignoniaceae ). These plants can cause acute cardiac failure often referred to as sudden death syndrome. The objective of this paper was to determine if N. stannea contains MFA and to report the clinical signs and pathology of cattle experimentally poisoned by this plant. MFA was detected in all N. stannea plant parts except mature leaves. Niedenzuella stannea was dosed to cattle at single doses of 15-30g kg-1 bw. Clinical signs and pathology observed were similar to those reported by other MFA-containing plants. Animals showed marked tachycardia, labored breathing, muscle tremors, loss of balance, ataxia, falling, recumbence, pedaling movements, and opisthotonous leading to death in few minutes. Gross lesions included engorgement of the large veins, severe lung edema, and petechiae , ecchymosis and suffusions in the pericardium, epicardium and pleura. Histologically, hydropic vacuolar degeneration and necrosis of epithelial cells of the kidney were observed. Cattle that received daily doses of 4g kg-1 body weight for 24 days showed no clinical signs. In summary, this is the first report that N. stannea contains monofluoroacetate and causes sudden death in cattle.

Pathology of Mycoplasma synoviae and other concurrent infections affected oviduct in commercial layer chicken

Pathology of oviduct affected with Mycoplasma synoviae and other concurrent infections was studied. Eggshell apex abnormalities (EAA) outbreak was reported in ten commercial layer farms in Namakkal district of Tamil Nadu which was suspected for M. synoviae infection. Oviduct samples were collected to study the pathology of oviduct and for the molecular confirmation of M. synoviae infection and other possible oviduct infections viz. Mycoplasma gallisepticum, E. coli and Infectious bronchitis virus by specific PCR. Out of ten farm samples screened eight were positive for M. synoviae, nine were positive for E. coli, all ten were positive for M. gallisepticum and all were negative for infectious bronchitis virus. On post-mortem examination, EAA affected birds exhibited no gross lesions except egg bound and congested oviduct in few cases. Microscopically, oviduct revealed degeneration, necrosis and desquamation of lining epithelial cells and atrophy of tubular glands. Serofibrinous exudates in the lumen and marked infiltration of macrophages and heterophils were also noticed. These findings showed that Eggshell apex abnormality producing oviducts had combined infection rather than individual etiology.

Ameliorative effects of pine bark extract on cisplatin-induced acute kidney injury in rats

Objective: This study investigated the dose–response effects of pine bark extract (PBE, pycnogenol®) on oxidative stress-mediated apoptotic changes induced by cisplatin (Csp) in rats.Materials and methods: The ameliorating potential of PBE was evaluated after orally administering PBE at doses of 10 or 20 mg/kg for 10 days. Acute kidney injury was induced by a single intraperitoneal injection of Csp at 7 mg/kg on test day 5.Results: Csp treatment caused acute kidney injury manifested by elevated levels of serum blood urea nitrogen (BUN) and creatinine (CRE) with corresponding histopathological changes, including degeneration of tubular epithelial cells, hyaline casts in the tubular lumen, and inflammatory cell infiltration (interstitial nephritis). Csp also induced significant apoptotic changes in renal tubular cells. In addition, Csp treatment induced high levels of oxidative stress, as evidenced by an increased level of malondialdehyde, depletion of the reduced glutathione (GSH) content, and decreased activities of glutathione S-transferase, superoxide dismutase, and catalase in kidney tissues. On the contrary, PBE treatment lowered BUN and CRE levels and effectively attenuated histopathological alterations and apoptotic changes induced by Csp. Additionally, treatment with PBE suppressed lipid peroxidation, prevented depletion of GSH, and enhanced activities of the antioxidant enzymes in kidney tissue.Conclusions: These results indicate that PBE has a cytoprotective effect against oxidative stress-mediated apoptotic changes caused by Csp in the rat kidney, which may be attributed to both increase of antioxidant enzyme activities and inhibition of lipid peroxidation.

PTEN Reduced UVB-Mediated Apoptosis in Retinal Pigment Epithelium Cells.

Age-related macular degeneration (AMD) is a leading cause of blindness and progressive loss of central vision in the elderly population. The important factor of AMD pathogenesis is the degeneration of retinal pigment epithelial (RPE) cells by oxidative stress. Inactivation of PTEN can disrupt intercellular adhesion in the RPE cells, but the mechanism of oxidative stress is less known. Here we presented evidence that UVB-mediated oxidative stress induced apoptosis in ARPE-19 cells. Downregulation of the expression of PTEN in UVB-irradiative RPE cells triggered DNA damage and increased the level of UVB-induced apoptosis by activating p53-dependent pathway. However, overexpression of PTEN increased cell survival by suppressing p-H2A in response to DNA damage and apoptosis. When using Pifithrin-α (one of p53 inhibitors), the level of p53-dependent apoptosis was significantly lower than untreated, which suggested that p53 was possibly involved in PTEN-dependent apoptosis. Thus, it elucidated the molecular mechanisms of UVB-induced damage in RPE cells and may offer an alternative therapeutic target in dry AMD.

Toxic proteins from Crotontiglium L. exert a proinflammatory effect by inducing release of proinflammatory cytokines and activating the p38-MAPK signaling pathway.

The aim of the present study was to determine the toxic targets of proteins from Croton tiglium L. and to investigate the potential mechanism of their toxicity. The toxic targets were determined by oral medication and intraperitoneal injection. The median lethal dose of oral medication in mice was calculated using Bliss software (2,752.8–3,407.5 mg/kg), and that of intraperitoneal injection was 195.8–272.69 mg/kg. The results of histopathological examination demonstrated that the kidney was primarily impaired by intraperitoneal injection, with slight degeneration of renal tubular epithelial cells. As to oral medication, the digestive tract was primarily injured, which manifested as congestion, bleeding, serious edema and other symptoms. Oral administration of the proteins caused gastrointestinal edema by increasing the intestinal permeability. Severe edema was associated with the inflammatory response, therefore the association between the toxicity of the proteins and inflammation was investigated. The proinflammatory effects of the crude proteins on the release of inflammatory mediator prostaglandin E2 (PGE2) were evaluated through intraperitoneal injection and the production of proinflammatory cytokines in RAW264.7 macrophages. Maximum PGE2 was released in the mice in vivo following intraperitoneal injection with 400 mg crude protein/kg body weight. Proinflammatory cytokines in macrophages, including tumor necrosis factor-α and interleukin-1β, were produced in dose- and time-dependent manners in vitro. furthermore, the expressions of cell signaling molecules were detected by western blotting. The inflammatory response induced by crude protein in macrophages was associated with the mitogen-activated protein kinase (MAPK) signaling pathway mainly including p38-MAPK, extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinase 1/2/3 and the activated p38-MAPK signaling pathway. However, extracellular signal-regulated kinase 1/2 and c-Jun N-terminal kinases 1–3 exhibited no significant response.

Protective effects of an HTRA1 insertion–deletion variant against age-related macular degeneration in the Chinese populations

Age-related macular degeneration (AMD) is a leading cause of visual impairment and irreversible blindness in most developed countries, affecting about 50 million elderly people worldwide. Retinal pigment epithelial (RPE) cell degeneration is the pathophysiological cause of AMD, leading to geographic atrophy and choroidal neovascularization. We and others have previously identified several polymorphisms on chromosome 10q26 (HTRA1 rs11200638 as well as LOC387715 rs10490924 and c.372_815del443ins54) associated with AMD. In this study, we confirmed the association of our previously identified HTRA1 insertion–deletion (indel) variant (c.34delCinsTCCT) in 195 exudative AMD patients and 390 controls from the Hong Kong Chinese cohort with additional 168 patients and 210 controls from the Chengdu Chinese cohort and followed by studying its biological functions in RPE cells. Genetic analysis verified the higher prevalence of c.34delCinsTCCT allele in control subjects (8.0%) than in AMD patients (1.9%; P=7.87 × 10−5, odds ratio=0.229). This protective effect was validated as the haplotype of the c.34delCinsTCCT allele existed independent of the risk haplotype (P=1.17 × 10−5). In vitro studies showed that recombinant HTRA1 c.34delCinsTCCT variant protein was more localized in the endoplasmic reticulum of RPE cells compared with the wild-type protein, and its secretion was delayed. Moreover, ARPE-19 cells expressing HTRA1 c.34delCinsTCCT variant had higher cell viability, lower cell apoptosis and were less responsive to anoikis, supporting its protective role. We revealed a protective AMD-associated HTRA1 variant in Chinese populations and the biological role of HTRA1 in RPE cell degeneration, indicating its involvement in AMD pathogenesis.

Pathological conditions of avian coccidiosis in the small scale commercial broiler farms in Dinajpur district

The study was designed to investigate the pathological conditions of avian coccidiosis in the small scale commercial broiler farms at different region in Dinajpur district during July, 2012 to December, 2012. A thorough clinical and necropsy examination was done and the characteristics clinical signs and gross lesions were recorded. Different organs mainly caecum and other parts of intestine were collected, preserved and processed for histopathological examination. Intestinal content was also examined for detection of oocyst. Total 234 diseased and dead birds (from 50 farms) were examined out of which 20 (8.54%) birds were found to be positive for coccidiosis. The clinical signs of the affected birds were bloody diarrhea, anemia, reduction of feed and water intake, drooping wings. At necropsy, enlargement and discoloration of caecum with numerous hemorrhagic spots, blood mixed and reddish to brown intestinal contents in the intestinal lumen, hemorrhage on the intestinal wall and mucosa were found. Histopathological examination revels distortion of normal architecture of intestine and desquamation of lining epithelia, formation of tissue debris on the intestinal mucosa and necrotic cells infiltration in the lamina propria and submucosa, degeneration of epithelial cells, glands and intestinal villi. So, outbreaks of coccidiosis in the commercial poultry flocks in Dinajpur district is lower due to farmers are intensely aware of coccidiosis now and they usually use coccidiostats routinely.Res. Agric., Livest. Fish.3(3): 425-431, December 2016

Histological Changes of Liver, Kidney and Brain in Uninephrectomized Male Rats Exposed to Fluoxetine

Kidney is the organ of humans and animals body which responsible for purification of blood from waste substances, the process of removing kidney or part of kidney called nephrectomy. The depression disease is mostly occurred in unhealthy person; it is commonly treated by fluoxetine. The purpose of this investigation to determine the histological effects of fluoxetine on liver, kidney, cerebrum and cerebellum of uninephrectomized male rats; rats were divided randomly into two groups, uninephrectomized control group and uninephrectomized fluoxetine treated group. After one month administration of fluoxetine, histological alteration observed in liver such as a little hydropic degeneration of the hepatocytes and inflammatory leucocyte infiltration. Hemorrhagic area and shrinking of glomerulus were appeared in the cortex region of kidney, degeneration of kidney tubule epithelial cells and dilation in some kidney tubules were detected in medulla region as a result of exposing uninephrectomized rats to fluoxetine. Histological slides of cerebrum and cerebellum uninephrectomized rats which exposed to fluoxetine showed dead pyramidal and purkinje cells respectively. In conclusion, light microscopic examinations showed that fluoxetine had a little effect on liver, kidney, cerebrum and cerebellum.

HISTOPATHOLOGICAL STUDIES OF SELECTED ORGANS OF OREOCHROMIS MOSSAMBICUS EXPOSED TO BIOPESTICIDE ACHOOK – SHORT TERM AND LONG TERM TOXICITY

The present study is aimed at evaluating the pathological changes in various tissues of Oreochromis mossambicus exposed to the neem based pesticide achook. The LC50 value obtained after 96 hrs was 0.105 ppm and 1/5th of LC50 value i.e., 0.021 ppm was taken as sublethal concentration. The fishes were exposed to this sublethal concentration for a period of 42-days. Fishes were dissected at the end of 7th day and 42nd day of exposure and the gill, liver and intestine were collected from exposed fishes and control group. Tissues were processed and sectioned at 4µm and stained with Haematoxylin-Eosin. The histological changes observed were mild on 7th day but became severe by 42nd day. The pathological changes in gills of exposed fishes include shrunken and fused gill lamellae, inflammatory cells in the primary and secondary gill lamellae. Exposed liver showed lesions consisting of vacuolar degeneration and disruption of hepatocytes. The changes found in intestine were vacuolar degeneration and disruption of epithelial cells of villi, ruptured and shapeless villi. The histology of tissues from control fishes were normal.

Acute renal injury induced by valacyclovir hydrochloride: A case report.

Acyclovir has been a frequently used antiviral agent in the clinical treatment of leukemia, acute encephalitis, malignant tumor and herpes simplex. The adverse effects of this drug have been widely described in clinical practice. In the present study, a case of a 35-year-old female patient diagnosed with herpes simplex, who developed acute renal injury following treatment with valacyclovir hydrochloride, is described. Kidney biopsy, light microscopy and laboratory examination were performed, and all findings revealed the signs of evident vacuolar degeneration of capillary endothelial and renal tubular epithelial cells, erythrocyte aggregation in partial renal tubule and microvilli exfoliation from epithelial cells. Renal interstitial edema was clearly identified. The clinical evidence observed from this female patient indicated that renal functions should be closely monitored during valacyclovir hydrochloride administration. A variety of effective measures, such as hydration, alkalizing urine, promoting the discharge of medication and the use of antagonists are recommended following the administration of antiviral agents.

In Vitro Angiogenesis: Co-Culture Model of the Retina in Microfluidic Devices

Background Age-related macular disease (AMD) is a common eye condition and the most severe form of a disease that is the leading cause of vision loss among people age 50 and above. Simple tasks such as driving or reading become impossible without a functional central vision, which is impaired as the result of damages to the macula. Wet form AMD is responsible for 90% of the blindness out of all AMD patients; it is marked by the development of subretinal choroidal neovascularization and degeneration of retinal pigmental epithelial (RPE) cells. Wet macular degeneration occurs when abnormal blood vessels disrupt the retina and leak blood or fluid that blurs the central vision. Recently, MEMS technology which integrates multidisciplinary field of studies offers a new direction in medical investigation and diagnosis. Here we utilized microfabrication technology to reconstruct ocular fundus tissues (OF) in vitro and to simulate neovascularization in the hope of building a disease model for the retina. Method Human retinal pigment epithelial (ARPE-19) and human umbilical vein endothelial cells (HUVECs), representing the retinal cells and the choroid, were co-cultured in the microfluidic devices. Two different approaches are developed in fabricating microfluidic devices resulting in 2D and 3D models. 2D model:the microfluidic device was made up of 2 layers, on each a channel (width 500 µm; height 150 µm) was embedded. A porous membrane (thickness 6.5 µm, hole diameter 10 µm) was sandwiched between the layers. Microchannels and membranes were made of poly(dimethylsiloxane) (PDMS). Retinal pigment epithelial (RPE) cells were introduced into the upper channel of the device and umbilical vein endothelial cells (HUVECs) were loaded into the lower channel. 3D model: the wire with a 0.2 mm diameter was pretreated before embedded in collagen (6.5 mg/ml). After the collagen had polymerized, the needle was removed resulting in an empty channel where HUVECs were to be seeded. RPE cells were seeded on the top of the collagen gel to initiate co-culture experiments. Data analysis: RPE cells and HUVEC were dyed respectively with Cell TrackerTM red and green and cell growth was recorded at certain time intervals. RPE cells were under hypoxia and hypoglycemia 24hr prior the start of the co-culture experiment. Before the aforementioned co-culture model, RPE were cultured alone on the upper channel and the VEGF concentration was collected and measured by ELIZA after replacing DME medium High Glucose (20 mM) with DME medium Low Glucose (3 mM). HUVECs were loaded into the lower channel and cultured alone. The upper channel was perfused with 50 ng/ml of VEGF-A165and the number of HUVECs migrating to the upper channel was analyzed using the same monitoring system. Characteristics of cells in the 3D model were characterized following the same methodology. Results Validation of the devices: The devices were validated prior culturing cells inside the devices. Confocal images of the cross-sections of channel length and diameter were taken after perfusion of fluorescent microbeads. ARPE characterization: Results show that RPE cells responded to the lowered glucose microenvironment by increasing VEGF secretion (58.8 % compared to the control). In addition, RPE cells exhibited a polar secretion of VEGF which is consistent to the cell physiology. HUVEC characterization: The increased number of HUVEC migrating through the porous membrane in response to VEGF gradient signifies neovascularization. Comparing to the control, there was a 31.3 % increase of cell migration. Conclusions and Future work The physiology of RPE cells and HUVECs is reproduced in the microfluidic devices. The preliminary results suggest that the alteration of the microenvironment induces responses of RPE cells and the significance of VEGF in neovascularization. In the near future, we anticipate studying responses of the co-culture model to hypoxia to characterize the AMD pathology.