SAMS-1 is required for the normal defecation motor program in Caenorhabditis elegans.

Defecation is an ultradian rhythmic behavior in Caenorhabditis elegans . We investigated the involvement of sams family genes in regulating the defecation motor program. We found that sams-1 mutants exhibited longer cycles than wild-type animals. With aging, the sams-1 mutants also frequently skipped the expulsion (Exp) step of defecation behavior. The sams-1 knockdown is known to reduce phosphatidylcholine (PC) levels, which are reversed by choline supplementation. We examined the effect of choline supplementation on defecation cycle times and Exp steps from adult days 1-4. Although choline supplementation did not alter the longer defecation cycle times of sams-1 mutants, it restored the loss of the Exp step in sams-1 mutants on adult days 3 and 4, suggesting a link between the regulation of the Exp step in sams-1 mutants and PC production.

Developmental adverse effects of trace amounts of lead: Evaluation using zebrafish model.

Lead (Pb) is widely used as a raw material for various daily necessities in human civilization. However, Pb is a major toxicant and Pb poisoning has long been a global health concern. A large body of evidence has revealed that exposure to Pb causes a variety of adverse health effects. Meanwhile, experimental studies on the developmental effects caused by trace amounts of Pb remain to be fully conducted. Therefore, we aimed to provide direct experimental evidence of the adverse developmental effects of Pb exposure below the occupational regulatory standard concentrations using a zebrafish model. We also attempted to investigate the cellular stress response caused by such a trace amount of Pb at the individual level. Fertilized zebrafish eggs were exposed to 100 ppb Pb from 6 to 72 h post fertilization (hpf), the developmental period included within the mammalian implantation to birth. The embryos exposed to Pb did not show superficially evident morphological alterations or differences in viability compared with the controls until 72 hpf; however, they hatched earlier and were significantly shorter in body length than the controls at 48 and 72 hpf. Larvae that were exposed to Pb until 72 hpf and then cultured until 7 days post fertilization without Pb exhibited edema and inflation defects in the swim bladder. The reactive oxygen species level in the Pb-exposed embryos was similar at 24 hpf, slightly but significantly higher at 48 hpf, and lower than half that of the control at 72 hpf. Accordingly, the expression levels of oxidative stress response-related genes were analyzed, and five out of seven tested genes were upregulated in Pb-exposed embryos at 48 and 72 hpf. In addition, the endoplasmic reticulum (ER) stress related genes were upregulated at 48 hpf. These results indicate that exposure of embryos to trace amounts of Pb induces a transient increase in oxidative- and ER-stresses and results in weak hypotrophy and subsequent abnormalities later in development. Our findings may be key to understanding the total health effects of Pb exposure, and indicate that the zebrafish model is suitable for the investigation of developmental toxicity of pollutants such as Pb.

N-acetylcysteine restores the cadmium toxicity of Caenorhabditis elegans.

Cadmium is a well-known environmental toxicant. At the cellular level, exposure to cadmium results in cytotoxic effects through the elevation of reactive oxygen species (ROS) production. Although cadmium exposure leads to the dysfunction of various organs, the underlying mechanisms of the toxic effects of cadmium in vivo are still largely unknown. Caenorhabditis elegans (C. elegans) is a useful model animal and exhibits unique biological reactions in response to environmental toxicants. In this study, the toxic mechanisms of cadmium exposure in C. elegans were investigated using N-acetylcysteine (NAC), which has dual functions, i.e., as a chelator of metals and as an antioxidant. NAC did not inhibit the uptake of cadmium into nematodes, suggesting that NAC did not function as a chelator of cadmium under these experimental conditions. Based on this finding, we investigated the effect of NAC as an antioxidant on representative phenotypic traits caused by cadmium exposure-reduced body length, aversion behavior, and shortened lifespan. NAC did not reverse the decreased body size but did clearly restore the aversion behavior and the shortened lifespan. These data suggest that aversion behavior and shortened lifespan are mediated by oxidative stress in C. elegans.

Intake of acrylamide at the dietary relevant concentration causes splenic toxicity in adult zebrafish.

Acrylamide (AA) has recently been recognized as an immediate hazardous chemical compound owing to its various toxicities and unavoidable contamination of certain daily foods prepared at a high temperature. AA in foods is thus a worldwide concern; however, its toxicity at the dietary relevant concentration has yet to be experimentally elucidated. To determine whether dietary AA intake causes adverse health effects, adult zebrafish were fed a diet containing AA at a relevant dose for one month. Although AA-fed zebrafish showed no superficial abnormalities, their spleen was severely swollen. Therefore, their spleen was analyzed histologically and pathologically and the changes in cytokine expression in their spleen were also examined. Based on our findings, the intake of AA-containing food caused splenic damages, including cyst formation, hemorrhage, and inflammation, which were accompanied by immune responses as indicated by the appearance of a melanomacrophage center, activation of macrophages, and upregulation of major inflammatory cytokines in the spleen. Collectively, for the first time, we provided experimental evidence of the splenic toxicity caused by dietary AA intake.

Integration of read-across and artificial neural network-based QSAR models for predicting systemic toxicity: A case study for valproic acid.

We present a systematic, comprehensive and reproducible weight-of-evidence approach for predicting the no-observed-adverse-effect level (NOAEL) for systemic toxicity by using read-across and quantitative structure-activity relationship (QSAR) models to fill gaps in rat repeated-dose and developmental toxicity data. As a case study, we chose valproic acid, a developmental toxicant in humans and animals. High-quality in vivo oral rat repeated-dose and developmental toxicity data were available for five and nine analogues, respectively, and showed qualitative consistency, especially for developmental toxicity. Similarity between the target and analogues is readily defined computationally, and data uncertainties associated with the similarities in structural, physico-chemical and toxicological properties, including toxicophores, were low. Uncertainty associated with metabolic similarity is low-to-moderate, largely because the approach was limited to in silico prediction to enable systematic and objective data collection. Uncertainty associated with completeness of read-across was reduced by including in vitro and in silico metabolic data and expanding the experimental animal database. Taking the "worst-case" approach, the smallest NOAEL values among the analogs (i.e., 200 and 100 mg/kg/day for repeated-dose and developmental toxicity, respectively) were read-across to valproic acid. Our previous QSAR models predict repeated-dose NOAEL of 148 (males) and 228 (females) mg/kg/day, and developmental toxicity NOAEL of 390 mg/kg/day for valproic acid. Based on read-across and QSAR, the conservatively predicted NOAEL is 148 mg/kg/day for repeated-dose toxicity, and 100 mg/kg/day for developmental toxicity. Experimental values are 341 mg/kg/day and 100 mg/kg/day, respectively. The present approach appears promising for quantitative and qualitative in silico systemic toxicity prediction of untested chemicals.

Blockade of ALK4/5 signaling suppresses cadmium- and erastin-induced cell death in renal proximal tubular epithelial cells via distinct signaling mechanisms.

Various types of cell death, including apoptosis, necrosis, necroptosis, and ferroptosis, are induced in renal tubular epithelial cells following exposure to environmental stresses and toxicants such as osmotic stress, ischemia/reperfusion injury, cisplatin, and cadmium. This is known to cause renal dysfunction, but the cellular events preceding stress-induced cell death in renal tubules are not fully elucidated. The activin receptor-like kinase (ALK) 4/5, also known as activin-transforming growth factor (TGF) ß receptor, is involved in stress-induced renal injury. We, therefore, studied the role of ALK4/5 signaling in HK-2 human proximal tubular epithelial cell death induced by cisplatin, cadmium, hyperosmotic stress inducer, sorbitol, and the ferroptosis activator, erastin. We found that ALK4/5 signaling is involved in cadmium- and erastin-induced cell death, but not sorbitol- or cisplatin-induced apoptotic cell death. Cadmium exposure elevated the level of phosphorylated Smad3, and treatment with the ALK4/5 kinase inhibitors, SB431542 or SB505124, suppressed cadmium-induced HK-2 cell death. Cadmium-induced cell death was attenuated by siRNA-mediated ALK4 or Smad3 silencing, or by treatment with SIS3, a selective inhibitor of TGFß1-dependent Smad3 phosphorylation. Furthermore, ALK4/5 signaling activated Akt signaling to promote cadmium-induced HK-2 cell death. In contrast, siRNA-mediated Inhibin-bA silencing or treatment with TGFß1 or activin A had little effect on cadmium-induced HK-2 cell death. On the other hand, treatment with SB431542 or SB505124 attenuated erastin-induced ferroptosis by hyperactivating Nrf2 signaling in HK-2 cells. These results suggest that blockade of ALK4/5 signaling protects against cadmium- and erastin-induced HK-2 cell death via Akt and Nrf2 signaling pathways, respectively.

In vitro and in vivo studies of oxidative stress responses against acrylamide toxicity in zebrafish.

Acrylamide (AA) is widely used in soil stabilization, water treatment, and industrial products and found in certain foods; however, its toxicity is an expanding global concern. Thus, to reveal the mechanisms involved in the development of, or protection from AA-induced toxicity has important significance. For this purpose, here we explored the intracellular stress response signaling pathways activated by AA exposure in zebrafish model. BRF41 cells derived from zebrafish were exposed to AA, and changes in the expression levels of 31 genes, including endoplasmic reticulum stress response-, oxidative stress response-, osmotic stress response-, and DNA damage and repair-related genes, were analyzed by PCR array. 12 genes upregulated in AA-exposed BRF41 cells were analyzed in zebrafish larvae by quantitative real time PCR, and the expression of all tested oxidative stress response-related genes was upregulated. Spatial expression patterns of these genes were visualized and found that their expression was upregulated and ectopically induced. In addition, AA-induced toxicity in BRF41 cells and the expression of glutathione S-transferase pi 1 (gstp1) in zebrafish larvae were reduced by N-acetylcysteine. Furthermore, inhibition of Gst activity enhanced AA toxicity. From these results, we concluded that the elicited oxidative stress response critically contributes to the protection from AA-induced toxicity.

Reduction of Intraocular Pressure by Additional Trabeculotomy Ab Externo in Eyes With Primary Open-angle Glaucoma.

PURPOSE: The purpose of this study was to determine whether a second trabeculotomy (LOT) can reduce the intraocular pressure (IOP) in eyes with primary open-angle glaucoma (POAG) that had undergone an unsuccessful LOT as the initial surgery. PATIENTS AND METHODS: LOT ab externo was performed as a second surgery on 37 eyes of 34 POAG patients who had undergone an unsuccessful LOT as the initial surgery. The main outcome measure was the postoperative IOPs, and surgical failures were defined as eyes with a post-LOT IOP>20 mm Hg. The eyes were divided into 3 groups; those that underwent LOT as both the initial and additional surgery (L-L group), those that underwent LOT as the initial surgery and combined LOT and cataract surgery (cLOT-IOL) as the additional surgery (L-cL group), and those that underwent cLOT-IOL as the initial surgery and LOT as the additional surgery (cL-L group). RESULTS: The IOP was reduced after the additional LOT at postoperative 24 months in the L-L group from 20.0±3.0 mm Hg to 15.3±2.6 mm Hg (P<0.001), the L-cL group from 19.8±1.6 mm Hg to 15.8±3.2 mm Hg (P=0.029), and the cL-L group from 20.1±2.7 mm Hg to 15.5±2.3 mm Hg (P=0.014). There were no differences in the preoperative and postoperative IOPs between the initial-operated and additional-operated eyes. The success rates were improved by the additional surgery in the L-L group (P<0.001) and the L-cL group (P=0.029), but the rate was worsened in the cL-L group (P<0.001). CONCLUSIONS: These results indicate that LOT is a reasonable choice as an additional glaucoma surgery after failure of an initial LOT.

Silver nanoparticles induce lysosomal-autophagic defects and decreased expression of transcription factor EB in A549 human lung adenocarcinoma cells.

Although silver nanoparticles (AgNPs) are widely used in consumer and medical products, the mechanism by which AgNPs cause pulmonary damage is unclear. AgNPs are incorporated into cells and processed via the autophagy pathway. We examined the effects of AgNP exposure on autophagic flux and expression of transcription factor EB (TFEB) in A549 lung adenocarcinoma cells. In cells exposed to citrate-coated 60-nm AgNPs, confocal laser microscopic examination showed a decrease in the LysoTracker fluorescence signal and an increase in that of Cyto-ID, indicating lysosomal pH alkalization and autophagosome formation, respectively. The proteins p62 and microtubule-associated protein light chain 3B-II (LC3B-II) are both degraded by autophagy, and their levels increased depending on AgNP dose. Furthermore, AgNP-induced increase in LC3B-II was not enhanced by treatment with the autophagic inhibitor bafilomycin A1. TFEB mRNA levels, and protein levels in cytosolic and nuclear fractions, were suppressed by exposure to AgNPs, suggesting transcriptional inhibition of TFEB expression. Overexpression of TFEB did not suppress AgNP-induced LC3B-II accumulation and cellular damage, indicating that impairment of autophagic flux and cellular damage by AgNPs might not be primarily caused by reduced TFEB expression. The present study suggests that AgNP-induced lysosomal dysfunction plays a principal role in the autophagic flux defect.

Age-dependent motor dysfunction due to neuron-specific disruption of stress-activated protein kinase MKK7.

c-Jun N-terminal kinase (JNK) is a member of the mitogen-activated protein kinase family and controls various physiological processes including apoptosis. A specific upstream activator of JNKs is the mitogen-activated protein kinase kinase 7 (MKK7). It has been reported that MKK7-JNK signaling plays an important regulatory role in neural development, however, post-developmental functions in the nervous system have not been elucidated. In this study, we generated neuron-specific Mkk7 knockout mice (MKK7 cKO), which impaired constitutive activation of JNK in the nervous system. MKK7 cKO mice displayed impaired circadian behavioral rhythms and decreased locomotor activity. MKK7 cKO mice at 8 months showed motor dysfunctions such as weakness of hind-limb and gait abnormality in an age-dependent manner. Axonal degeneration in the spinal cord and muscle atrophy were also observed, along with accumulation of the axonal transport proteins JNK-interacting protein 1 and amyloid beta precursor protein in the brains and spinal cords of MKK7 cKO mice. Thus, the MKK7-JNK signaling pathway plays important roles in regulating circadian rhythms and neuronal maintenance in the adult nervous system.

Involvement of Notch1 signaling in malignant progression of A549 cells subjected to prolonged cadmium exposure.

Cadmium exposure is known to increase lung cancer risk, but the underlying molecular mechanisms in cadmium-stimulated progression of malignancy are unclear. Here, we examined the effects of prolonged cadmium exposure on the malignant progression of A549 human lung adenocarcinoma cells and the roles of Notch1, hypoxia-inducible factor 1α (HIF-1α), and insulin-like growth factor 1 receptor (IGF-1R)/Akt/extracellular signal-regulated kinase (ERK)/p70 S6 kinase 1 (S6K1) signaling pathways. Exposing A549 cells to 10 or 20 µm cadmium chloride (CdCl2) for 9-15 weeks induced a high proliferative potential, the epithelial-mesenchymal transition (EMT), stress fiber formation, high cell motility, and resistance to antitumor drugs. Of note, the CdCl2 exposure increased the levels of the Notch1 intracellular domain and of the downstream Notch1 target genes Snail and Slug. Strikingly, siRNA-mediated Notch1 silencing partially suppressed the CdCl2-induced EMT, stress fiber formation, high cell motility, and antitumor drug resistance. In addition, we found that prolonged CdCl2 exposure induced reduction of E-cadherin in BEAS-2B human bronchial epithelial cells and antitumor drug resistance in H1975 human tumor-derived non-small-cell lung cancer cells depending on Notch1 signaling. Moreover, Notch1, HIF-1α, and IGF-1R/Akt/ERK/S6K1 activated each other to induce EMT in the CdCl2-exposed A549 cells. These results suggest that Notch1, along with HIF-1α and IGF-1R/Akt/ERK/S6K1 signaling pathways, promotes malignant progression stimulated by prolonged cadmium exposure in this lung adenocarcinoma model.

Application of Zebrafish Model to Environmental Toxicology.

Recently, a tropical freshwater fish, the zebrafish, has been generally used as a useful model organism in various fields of life science worldwide. The zebrafish model has also been applied to environmental toxicology; however, in Japan, it has not yet become widely used. In this review, we will introduce the biological and historical backgrounds of zebrafish as an animal model and their breeding. We then present the current status of toxicological experiments using zebrafish that were treated with some important environmental contaminants, including cadmium, organic mercury, 2,3,7,8-tetrachlorodibenzo-p-dioxin, and tributyltin. Finally, the future possible application of genetically modified zebrafish to the study of environmental toxicology is discussed.

Endoplasmic reticulum stress-mediated neuronal apoptosis by acrylamide exposure.

Acrylamide (AA) is a well-known neurotoxic compound in humans and experimental animals. However, intracellular stress signaling pathways responsible for the neurotoxicity of AA are still not clear. In this study, we explored the involvement of the endoplasmic reticulum (ER) stress response in AA-induced neuronal damage in vitro and in vivo. Exposure of SH-SY5Y human neuroblastoma cells to AA increased the levels of phosphorylated form of eukaryotic translation initiation factor 2α (eIF2α) and its downstream effector, activating transcription factor 4 (ATF4), indicating the induction of the unfolded protein response (UPR) by AA exposure. Furthermore, AA exposure increased the mRNA level of c/EBP homologous protein (CHOP), the ER stress-dependent apoptotic factor, and caused the accumulation of reactive oxygen species (ROS) in SH-SY5Y cells. Treatments of SH-SY5Y cells with the chemical chaperone, 4-phenylbutyric acid and the ROS scavenger, N-acetyl-cysteine reduced the AA-induced expression of ATF4 protein and CHOP mRNA, and resulted in the suppression of apoptosis. In addition, AA-induced eIF2α phosphorylation was also suppressed by NAC treatment. In consistent with in vitro study, exposure of zebrafish larvae at 6-day post fertilization to AA induced the expression of chop mRNA and apoptotic cell death in the brain, and also caused the disruption of brain structure. These findings suggest that AA exposure induces apoptotic neuronal cell death through the ER stress and subsequent eIF2α-ATF4-CHOP signaling cascade. The accumulation of ROS by AA exposure appears to be responsible for this ER stress-mediated apoptotic pathway.

Fluoride-induced c-Fos expression in MC3T3-E1 osteoblastic cells.

Excessive systemic exposure to fluoride leads to disturbances of bone homeostasis. c-Fos is known to be essential in bone development by affecting osteoblast and osteoclast differentiation. In this study, we examined the effects of fluoride exposure on c-Fos expression and its regulatory signaling pathways in MC3T3-E1 mouse osteoblast cell line. c-fos mRNA level, c-Fos protein level and c-Fos DNA-binding activity were markedly increased, with a peak at 2 or 4 h, in MC3T3-E1 cells exposed to sodium fluoride (NaF). Fra-1 protein, another member of Fos family, was also elevated, whereas FosB and Fra-2 proteins remained unchanged. NaF further induced phosphorylation of mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated protein kinase 1/2 (ERK1/2), ERK5, c-Jun NH2-terminal kinase and p38. NaF-induced expression of c-Fos protein was markedly suppressed with U0126, the inhibitor of both activated and non-activated forms of MAPK/ERK kinase 1/2 (MEK1/2) and BIX02189, the MEK5 inhibitor, but partially with SP600125, the JNK inhibitor and SB203580, the p38 inhibitor. Therefore, ERK1/2 and ERK5 signal transduction pathways are important for accumulating c-Fos. siRNA targeting against the mouse c-fos gene further enhanced NaF-induced up-regulation of osteoprotegerin (OPG), an inhibitor of osteoclastogenesis, suggesting that c-Fos might negatively regulate OPG expression induced by fluoride in osteoblastic cells.

Involvement of lysosomal dysfunction in silver nanoparticle-induced cellular damage in A549 human lung alveolar epithelial cells.

BACKGROUND: While silver nanoparticles (AgNPs) are widely used in consumer and medical products, the mechanism by which AgNPs cause pulmonary cytotoxicity is not clear. AgNP agglomerates are found in endo-lysosomal structures within the cytoplasm of treated cells. In this study, the functional role of lysosomes in AgNP-induced cellular damage was examined in A549 human lung alveolar epithelial cells. We evaluated the intracellular distribution of AgNPs, lysosomal pH, cellular viability, Ag dissolution, and metallothionein (MT) mRNA levels in AgNP-exposed A549 cells that were treated with bafilomycin A1, the lysosomal acidification inhibitor. FINDINGS: Exposure of A549 cells to citrate-coated AgNPs (20 nm diameter) for 24 h induced cellular damage and cell death at 100 and 200 µg Ag/ml, respectively. Confocal laser microscopic examination of LysoTracker-stained cells showed that AgNPs colocalized with lysosomes and their agglomeration increased in a dose-dependent manner (50-200 µg Ag/ml). In addition, the fluorescence signals of LysoTracker were reduced following exposure to AgNPs, suggesting the elevation of lysosomal pH. Treatment of A549 cells with 200 nM bafilomycin A1 and AgNPs (50 µg Ag/ml) further reduced the fluorescence signals of LysoTracker. AgNP-induced cell death was also increased by bafilomycin A1 treatment. Finally, treatment with bafilomycin A1 suppressed the dissolution of Ag and decreased the mRNA expression levels of MT-I and MT-II following exposure to AgNPs. CONCLUSIONS: The perturbation of lysosomal pH by AgNP exposure may play a role in AgNP agglomeration and subsequent cellular damage in A549 cells.

Experimental Evidence Shows Salubrinal, an eIF2α Dephosphorylation Inhibitor, Reduces Xenotoxicant-Induced Cellular Damage.

Accumulating evidence indicates that endoplasmic reticulum (ER) stress and the subsequent unfolded protein response (UPR) are involved in the pathogenesis of not only the protein misfolding disorders such as certain neurodegenerative and metabolic diseases, but also in the cytotoxicity of environmental pollutants, industrial chemicals, and drugs. Thus, the modulation of ER stress signaling pathways is an important issue for protection against cellular damage induced by xenotoxicants. The substance salubrinal has been shown to prevent dephosphorylation of the eukaryotic translation initiation factor 2 alpha (eIF2α). The phosphorylation of eIF2α appears to be cytoprotective during ER stress, because inhibition of the translation initiation activity of eIF2α reduces global protein synthesis. In addition, the expression of activating transcription factor 4 (ATF4), a transcription factor that induces the expression of UPR target genes, is up-regulated through alternative translation. This review shows that salubrinal can protect cells from the damage induced by a wide range of xenotoxicants, including environmental pollutants and drugs. The canonical and other possible mechanisms of cytoprotection by salubrinal from xenotoxicant-induced ER stress are also discussed.

Case of Primary Intraocular Lymphoma with Extraocular Extension.

PURPOSE: To describe a case of primary intraocular lymphoma (PIOL) with an extension through the sclera that was confirmed to be part of the PIOL by histopathological examinations. CASE: An 89-year-old woman was referred to a local clinic with a 1-year history of persistent blurred vision in her left eye. After 2 years without aggressive treatments, she had a marked reduction of vision and pain in her left eye. The clinical diagnosis was panophthalmitis, and the eye was enucleated and submitted for histopathological study. RESULTS: Light microscope examination showed that atypical lymphocytic cells had infiltrated into both the intraocular and extraocular areas. The anterior chamber angle was blocked by infiltrating tumor cells, which were also detected around the optic nerve. The tumor cells destroyed Bruch's membrane and infiltrated around the perineural and perivascular areas within the sclera. Immunohistochemistry showed that the tumor cells were positive for B-lymphocyte surface antigen (CD20), B-cell antigen receptor complex-associated protein alpha chain (CD79-alpha), and had a high positive rate for anti-Ki-67 antibody. CONCLUSION: The finding in our case indicates that early diagnosis and treatment are important for eyes with PIOL because the tumor can spread and penetrate the sclera and invade extraocular tissues.

Favorable anatomic and visual outcomes with 25-gauge vitrectomy for myopic foveoschisis.

PURPOSE: To report the surgical outcomes of 25-gauge vitrectomy in eyes with myopic foveoschisis (MF). METHODS: The medical records of 40 eyes of 36 patients that had undergone 25-gauge vitrectomy with internal limiting membrane peeling for MF were studied. The main outcome measures were the best-corrected visual acuity (BCVA) and the optical coherence tomography (OCT) findings. The eyes were divided into two groups: 1) those with a foveal detachment (FD; FD group); and 2) those without a FD (no-FD group). RESULTS: The postoperative OCT images showed a resolution of the MF with a significant reduction in the central foveal thickness from the preoperative values in both the FD group (479±150 µm to 196±56 µm; P=0.002, mean ± standard deviation) and in the no-FD group (369±116 µm to 245±50 µm; P=0.001). The final mean BCVA significantly improved from the preoperative values in the FD group (0.96±0.53 logarithm of the minimum angle of resolution [logMAR] units to 0.70±0.56 logMAR units; P=0.009) and in the no-FD group (0.46±0.38 logMAR units to 0.34±0.36 logMAR units; P=0.007). The final BCVA in the FD group improved in 63%, remained unchanged in 31%, and worsened in 6%. In the no-FD group, the final BCVA improved in 21%, remained unchanged in 71%, and worsened in 8% of the eyes. A better final BCVA was significantly correlated with a better preoperative BCVA in both groups (P<0.001). CONCLUSION: Twenty five-gauge vitrectomy results in favorable visual and anatomic outcomes for MF. We recommend that 25-gauge vitrectomy be used to treat eyes with MF.

Increased expression and activation of serum- and glucocorticoid-inducible kinase-1 (SGK1) by cadmium in HK-2 renal proximal tubular epithelial cells.

In HK-2 cells exposed to cadmium chloride (CdCl2), the level of serum- and glucocorticoid-inducible kinase-1 (SGK1) protein is increased, but the levels of SGK2 and SGK3 proteins are not. Phosphorylation of SGK1 protein is also observed. Treatment with actinomycin D abolished CdCl2-induced elevation of SGK1 mRNA level. Treatment with actinomycin D or cycloheximide suppressed SGK1 protein levels in cells exposed to CdCl2. Treatment with SGK1 inhibitor EMD638683 or knockdown of SGK1 with siRNA suppressed CdCl2-induced phosphorylation of N-Myc downstream-regulated kinase 1 (NDRG1). These results indicate that cadmium induces the transcriptional upregulation of SGK1 expression and regulates NDRG1 in HK-2 cells.

Clinical features of diabetic patients referred by general physicians due to less ophthalmic examinations.

BACKGROUND: The purpose of this study was to determine the clinical features of patients with type 2 diabetes, and less ophthalmic examinations, referred by general physicians to ophthalmologists. METHODS: The medical charts of 327 patients with type 2 diabetes referred to our department from general physicians were reviewed. A detailed medical history was taken and a complete ophthalmic examination was performed for all patients. The patients were divided into two groups, ie, those with a history of missing ophthalmic examinations for more than a year (noncompliant group) and those with no previous ophthalmic examinations (never-examined group). Serum levels of glycosylated hemoglobin and creatinine, estimated glomerular filtration rate, and urine albumin/creatinine ratio were obtained from medical records. RESULTS: Of the 327 patients, 102 had diabetic retinopathy (31.2%), with a mean best-corrected visual acuity of 0.037±0.36 logMAR (logarithm of the minimum angle of resolution) units. Of the 327 patients, 203 were in the never-examined group and 124 were in the noncompliant group. The incidence of diabetic retinopathy in the noncompliant group was significantly higher than that in the never-examined group (P<0.001). Best-corrected visual acuity in the noncompliant group was significantly worse than in the never-examined group (P=0.004). Glycosylated hemoglobin levels and estimated glomerular filtration rate in the noncompliant group were significantly lower than in the never-examined group (P<0.001 and P<0.003, respectively); serum creatinine levels and urine albumin/creatinine ratio were significantly higher (P=0.020 and P=0.001, respectively). The severity of the diabetic retinopathy was significantly correlated with compliance in terms of ophthalmic examinations and with urine albumin/creatinine ratio (multiple regression analysis, P=0.047 and P<0.001, respectively). CONCLUSION: Our results show that diabetic patients referred from general physicians due to less ophthalmic examinations generally have good visual acuity, but one third of them have diabetic retinopathy. A history of missing ophthalmic examinations and albuminuria are risk factors for diabetic retinopathy.