BACKGROUND AND OBJECTIVES: In regenerative medicine, mesenchymal stem cells derived from adipose tissues (Ad-MSCs) are a very attractive target to treat many diseases. In relation to nephrology, the aim of the current study is to investigate the effects of Ad-MSCs for the amelioration of acute kidney injury and to explore the mechanism of renal parenchymal changes in response to allogeneic transplantation of Ad-MSCs. METHODS AND RESULTS: The nephrotoxicity was induced by cisplatin (CP) in balb/c mice according to RIFLE Class and AKIN Stage 3. PCR, qRT-PCR and fluorescent labeled cells infusion, histopathology, immunohistochemistry, functional analyses were used for genes and proteins expressions data acquisition respectively. We demonstrated that single intravenous infusion of 2.5×107/kg mAd-MSCs in mice pre-injected with CP recruited to the kidney, restored the renal structure, and function, which resulted in progressive survival of mice. The renal tissue morphology was recovered in terms of diminished necrosis or epithelial cells damage, protein casts formation, infiltration of inflammatory cells, tubular dilatation, and restoration of brush border protein; Megalin and decreased Kim-1 expressions in mAd-MSCs transplanted mice. Significant reduction in serum creatinine with slashed urea and urinary protein levels were observed. Anti-BrdU staining displayed enhanced tubular cells proliferation. Predominantly, downgrade expressions of TNF-α and TGF-β1 were observed post seven days in mAd-MSCs transplanted mice. CONCLUSIONS: Ad-MSCs exerts pro-proliferative, anti-inflammatory, and anti-fibrotic effects. Ad-MSCs transplantation without any chemical or genetic manipulation can provide the evidence of therapeutic strategy for the origin of regeneration and overall an improved survival of the system in functionally deprived failed kidneys.
Acute Kidney Injury ; Animals ; Cisplatin ; Creatinine ; Dilatation ; Epithelial Cells ; Immunohistochemistry ; Infusions, Intravenous ; Kidney ; Low Density Lipoprotein Receptor-Related Protein-2 ; Mesenchymal Stromal Cells ; Mice ; Microvilli ; Necrosis ; Nephrology ; Polymerase Chain Reaction ; Regeneration ; Regenerative Medicine ; Transplantation, Homologous ; Urea

The vomeronasal organ has an important role in mammal's social and sexual behaviours. In addition, it mediates defensive behavior through detection of protein pheromone homologues. In this work, a detailed morphological description of the postnatal development of the non-sensory epithelium (NSE) lining the vomeronasal duct (VND) of the female cat is provided using various histological techniques. The study focused on newborn, 2 weeks, 4 weeks, and 8 weeks of postnatal ages using four animals for each age. We report here for the first time that three types of NSE line the rostral segment of the VND; nonkeratinized stratified squamous epithelium, stratified cuboidal epithelium, and ciliated pseudo stratified columnar ciliated epithelium with goblet cells and that the VND undergoes 90° a change in its its axis from the vertical position caudally to the horizontal position rostral. The NSE which lines the lateral side of the VND middle segment is consists of cliated pseudostratified columnar epithelium without goblet cells. In addition to basal cells, the NSE contains ciliated and three types of nonciliated columnar epithelial cells (dark, light, and unstained). Mitotic figures were observed only in the basal cells layer during the first 2 weeks of postnatal development. Intraepithelial invading inflammatory cells were uncommon. Scanning electron microscopy revealed unevenly distributed long cilia intermingled with microvillar processes and intervening short microvillar processes. These projecting cilia and microvilli revealed a gradual increase in their height during development toward maturity.
Animals ; Cats* ; Cilia ; Epithelial Cells ; Epithelium* ; Female ; Goblet Cells ; Histological Techniques ; Humans ; Infant, Newborn ; Microscopy, Electron, Scanning ; Microvilli ; Vomeronasal Organ*

Four thoroughbred horses showing lameness, ataxia, circling, depression, recumbency, and seizures, were examined. The horses had gross, pale- to dark-red manifestations and foci in the central nervous system (CNS). Multifocal to coalescing eosinophilic necrotizing encephalomyelitis was observed histologically in the CNS along with intact or degenerated nematodes. Nematodes had polymyarian-coelomyarian musculature, a smooth thin cuticle, and intestines lined by multinucleated cells with microvilli. These traits suggested the nematodes belonged to the family Protostrongylidae, which includes Parelaphostrongylus tenuis. It was concluded that the horses were infected by nematodes, presumably Parelaphostrongylus tenuis, resulting in eosinophilic necrotizing encephalomyelitis.
Ataxia ; Central Nervous System ; Depression ; Encephalomyelitis* ; Eosinophils* ; Horses* ; Humans ; Intestines ; Microvilli ; Parasites* ; Seizures

Microvillus inclusion disease (MVID) is an autosomal recessive disorder caused by biallelic mutations in the MYO5B or STX3 gene. Refractory diarrhea and malabsorption are the main clinical manifestations. The aim of this study was to investigate the clinical features and MYO5B gene mutations of an infant with MVID. A 21-day-old female infant was referred to the hospital with the complaint of diarrhea for 20 days. On physical examination, growth retardation of the body weight and length was found along with moderately jaundiced skin and sclera. Breath sounds were clear in the two lungs and the heart sounds were normal. The abdomen was distended and the veins in the abdominal wall were observed. The liver and spleen were not palpable. Biochemical analysis revealed raised serum total bile acids, bilirubin, transaminases and γ-glutamyl transpeptidase while decreased levels of serum sodium, chloride, phosphate and magnesium. Blood gas analysis indicated metabolic acidosis. The preliminary diagnosis was congenital diarrhea, and thus parenteral nutrition was given along with other symptomatic and supportive measures. However, diarrhea, metabolic acidosis and electrolyte disturbance were intractable, and the cholestatic indices, including transaminases, γ-glutamyl transpeptidase, bilirubin and total bile acids, remained at increased levels. One month later, the patient was discharged and then lost contact. On genetic analysis, the infant was proved to be a compound heterozygote of the c.310+2Tdup and c.1966C>T(p.R656C) variants of the gene MYO5B, with c.310+2Tdup being a novel splice-site mutation. MVID was thus definitely diagnosed.
Female ; Humans ; Infant, Newborn ; Malabsorption Syndromes ; diagnosis ; genetics ; Microvilli ; genetics ; pathology ; Mucolipidoses ; diagnosis ; genetics ; Mutation ; Myosin Heavy Chains ; genetics ; Myosin Type V ; genetics

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.
Cell Proliferation ; Chromatin ; Dry Eye Syndromes ; Epithelial Cells* ; Humans* ; Hyaluronic Acid* ; L-Lactate Dehydrogenase ; Microscopy ; Microscopy, Electron ; Microvilli ; Vacuoles ; Wound Healing* ; Wounds and Injuries*

OBJECTIVETo explore the clinical features and mutations of MYO5B gene in a family affected with microvillus inclusion disease.

METHODSClinical data of an infant affected with microvillus inclusion disease was collected. Genomic DNA was extracted from peripheral blood samples from the patient and her parents. PCR amplification and Sanger sequencing were performed to analyze all the exons and their flanking sequences of the MYO5B gene.

RESULTSThe patient presented with complicated manifestations including respiratory distress syndrome, dehydration, acidosis, bowel dilatation, liver and kidney dysfunction, and severe and intractable diarrhea. A compound mutation of the MYO5B gene, i.e., IVS37-1G>C/c.2729_2731delC (p.R911Afs916X), was discovered in the patient. The former was a splice-site mutation inherited from the mother, while the latter was a frameshift mutation inherited from the father. Both were not reported previously.

CONCLUSIONBased on the clinical and molecular evidence, the patient was diagnosed with microvillus inclusion disease. Above finding has expanded the mutation spectrum of the MYO5B gene, which can provide valuable information for genetic counseling for the family.

Family ; Female ; Genetic Testing ; methods ; Genotype ; Humans ; Infant ; Malabsorption Syndromes ; genetics ; Male ; Microvilli ; genetics ; pathology ; Mucolipidoses ; genetics ; Mutation ; genetics ; Myosin Heavy Chains ; genetics ; Myosin Type V ; genetics ; Phenotype

PURPOSE: To investigate the biological effects of preservative-free artificial tear drops on cultured human corneal epithelial cells in vitro. METHODS: The effects of the preservative-free artificial tear drops (Kynex(R) 0.1%, Kynex II(R) 0.18% [Alcon, Seoul, Korea] and Hyaluni eye drops(R) 0.15%, 0.3% [Taejun, Seoul, Korea]) on the human corneal epithelial cells were evaluated. An methyl thiazolyl tetrazolium (MTT)-based colorimetric assay was performed to assess the cellular metabolic activity and a lactate dehydrogenase (LDH) leakage assay was used to determine cellular toxicity. The eye drop ingredients were analyzed for electrolyte composition, pH, and osmolarity. We performed a scratch assay and cellular morphology test using electronic microscopy. RESULTS: The metabolic activity of corneal epithelial cells was higher than controls at 24 hours after exposure and then decreased at 48 and 72 hours after exposure (p < 0.05). The LDH titers of the 4 eye drops were higher compared with controls (p < 0.05). Sodium hyaluronate 0.18% contained lower concentrations of Na+ or Cl- and showed lower osmolarity values compared with the other eye drops. The cellular migration based on the scratch assay was more delayed and cellular damage such as loss of microvilli, rough endothelial reticulum (RER), and mitochondria dilatation was greater than controls based on electron microscopy. CONCLUSIONS: Long-term exposure to preservative-free sodium hyaluronate eye drops may induce decreased metabolic activity and cellular damage. Thus, preservative-free artificial tears should be used carefully to prevent cellular toxicity.
Cornea ; Dilatation ; Epithelial Cells* ; Epithelium ; Humans* ; Hyaluronic Acid* ; Hydrogen-Ion Concentration ; L-Lactate Dehydrogenase ; Microscopy ; Microscopy, Electron ; Microvilli ; Mitochondria ; Ophthalmic Solutions ; Osmolar Concentration ; Reticulum ; Seoul ; Sodium* ; Tears

It is well known that proteins present in the primary urine are reabsorbed in the renal proximal tubules, and that this reabsorption is mediated via the megalincubilin complex and the neonatal Fcgamma receptor. However, the reabsorption is also thought to be influenced by an electrostatic interaction between protein molecules and the microvilli of the renal proximal tubules. By analyzing the charge diversity of urinary IgG, we showed that this reabsorption process occurs in a cationic charge-preferential manner. The charge-selective molecular sieving function of the glomerular capillary walls has long been a target of research since Brenner et al. demonstrated the existence of this function by a differential clearance study by using the anionic dextran sulfate polymer. However, conclusive evidence was not obtained when the study was performed using differential clearance of serum proteins. We noted that immunoglobulin (Ig) A and IgG have similar molecular sizes but distinct molecular isoelectric points. Therefore, we studied the differential clearance of these serum proteins (clearance IgA/ clearance IgG) in podocyte diseases and glomerulonephritis. In addition, we studied this differential clearance in patients with Dent disease rather than in normal subjects because the glomerular sieving function is considered to be normal in subjects with Dent disease. Our results clearly showed that the charge-selective barrier is operational in Dent disease, impaired in podocyte disease, and lacking in glomerulonephritis.
Blood Proteins ; Capillaries ; Child Health ; Dent Disease ; Dextran Sulfate ; Glomerulonephritis ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulins ; Isoelectric Point ; Microvilli ; Nephritis ; Podocytes ; Polymers ; Proteinuria*

OBJECTIVETo observe the ultrastructural change of the route of gut bacterial translocation in a rat with spinal cord injury (SCI).

METHODSForty Wistar rats were divided into the following groups: control group and 3 SCI groups (10 in each group). The rats in the SCI groups were established SCI model at 24 h, 48 h, and 72 h after SCI. Small intestine mucous membrane tissue was identified and assayed by transmission electron microscope, scanning electron microscope and immunofluorescence microscopy.

RESULTSSmall intestine mucous membrane tissue in control group was not damaged significantly, but those in SCI groups were damaged significantly. Proliferation bacteria in gut lumen attached on microvilli. The extracellular bacteria torn the intestinal barrier and perforated into the small intestinal mucosal epithelial cell. The bacteria and a lot of particles of the seriously damaged region penetrated into the lymphatic system and the blood system directly. Some bacteria were internalized into the goblet cell through the apical granule. Some bacteria and particles perforated into the submucosa of the M cell running the long axis of M cells through the tight junctions. In the microcirculation of mucosa, the bacteria that had already broken through the microvilli into blood circulation swim accompanying with erythrocytes.

CONCLUSIONThe routes of bacterial translocation interact and format a vicious circle. At early step, the transcellular pathway of bacterial translocation is major. Following with the destroyed small intestine mucous, the routes of bacterial translocation through the lymphatic system and the blood system become direct pathways. The goblet cell-dendritic cell and M cell pathway also play an important role in the bacterial translocation.

Animals ; Bacteria ; Bacterial Translocation ; Epithelial Cells ; microbiology ; Goblet Cells ; microbiology ; Intestinal Mucosa ; microbiology ; pathology ; ultrastructure ; Intestine, Small ; microbiology ; pathology ; ultrastructure ; Microvilli ; microbiology ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; microbiology

Objective of present study was to prepare and characterize self-nanoemulsifying drug delivery system (SNEDDS) of lutein and to evaluate its effect on bioavailability of warfarin. The SNEDDS was prepared using an oil, a surfactant, and co-surfactants with optimal composition based on pseudo-ternary phase diagram. Effect of the SNEDDS on the bioavailability of warfarin was performed using Sprague Dawley rats. Lutein was successfully formulated as SNEDDS for immediate self-emulsification and dissolution by using combination of Peceol as oil, Labrasol as surfactant, and Transcutol-HP or Lutrol-E400 as co-surfactant. Almost complete dissolution was achieved after 15 min while lutein was not detectable from the lutein powder or intra-capsule content of a commercial formulation. SNEDDS formulation of lutein affected bioavailability of warfarin, showing about 10% increase in Cmax and AUC of the drug in rats while lutein as non-SNEDDS did not alter these parameters. Although exact mechanism is not yet elucidated, it appears that surfactant and co-surfactant used for SNEDDS formulation caused disturbance in the anatomy of small intestinal microvilli, leading to permeability change of the mucosal membrane. Based on this finding, it is suggested that drugs with narrow therapeutic range such as warfarin be administered with caution to avoid undesirable drug interaction due to large amount of surfactants contained in SNEDDS.
Animals ; Area Under Curve ; Biological Availability* ; Drug Delivery Systems* ; Drug Interactions ; Lutein* ; Membranes ; Microvilli ; Permeability ; Rats ; Rats, Sprague-Dawley ; Surface-Active Agents ; Warfarin*