Intraocular pharmacokinetics of intravitreal vascular endothelial growth factor-Trap in a rabbit model.

PURPOSE: To determine intraocular pharmacokinetic properties of intravitreally injected vascular endothelial growth factor (VEGF)-Trap in a rabbit model. METHODS: VEGF-Trap was intravitreally injected in 18 rabbit eyes. Eyes were enucleated 1 h and 1, 2, 5, 14, and 30 days after injections and immediately frozen at -80 °C. Concentration of VEGF-Trap in vitreous, aqueous humor, and retina/choroid was determined using an indirect enzyme-linked immunosorbent assay and analyzed to obtain pharmacokinetic properties. RESULTS: Maximum concentration of VEGF-Trap was achieved at 1 h in all three tissues. A one-compartment model of distribution was selected as the final model for all tissues studied. Estimated half-life of VEGF-Trap in vitreous, aqueous humor, and retinal/choroid was 87.1, 36.8, and 35.0 h, respectively, and estimated mean residence time was 125.7, 53.1, and 50.5 h, respectively. Area under the curve from time 0 to the end point was 10009.8, 3945.1, and 1189.3, respectively. Total exposure of the aqueous humor and retina/choroid to VEGF-Trap was 39.4% and 11.9% of vitreous exposure, respectively. CONCLUSION: The vitreous half-life of VEGF-Trap is 3.63 days. This is shorter than that of bevacizumab (6.99 days) and longer than that of ranibizumab (2.51 days), as shown in studies using the same experimental settings. The concentration of VEGF-Trap peaked at 1 h after injections in all eye tissues studied.

Circadian disruption in the pathogenesis of metabolic syndrome.

Metabolic syndrome is a multifactorial process induced by a combination of genetic and environmental factors and recent evidence has highlighted that circadian disruption and sleep loss contribute to disease pathogenesis. Emerging work in experimental genetic models has provided insight into the mechanistic basis for clock disruption in disease. Indeed, disruption of the clock system perturbs both neuroendocrine pathways within the hypothalamus important in feeding and energetics, in addition to peripheral tissues involved in glucose and lipid metabolism. This review illustrates the impact of molecular clock disruptions at the level of both brain and behavior and peripheral tissues, with a focus on how such dysregulation in turn impacts lipid and glucose homeostasis, inflammation and cardiovascular function. New insight into circadian biology may ultimately lead to improved therapeutics for metabolic syndrome and cardiovascular disease in humans.

Stem-cell therapy for peripheral arterial occlusive disease.

OBJECTIVE: The aims of our study were to confirm the effectiveness via animal study and safety through clinical trials of using human cord blood-mononuclear cells (HCB-MNCs). DESIGN: We performed a dose-response animal study (HCB-MNCs: 4 × 106, 4 × 107 and 4 × 108) using a limb ischaemia model in dogs to assess angiogenic responses. Safety assessment in humans in terms of graft-versus-host-disease was also done by observing an uncontrolled case series. MATERIALS AND METHODS: Twelve animal ischaemic limbs and seven patients with thromboangiitis obliterans were treated with HCB-MNCs. These cells (4 × 108) were injected into the ischaemic limb muscle of patients. The results were analysed at 8 weeks for the animal study and at 6 months for patients. RESULTS: In the animal ischaemic models, the number of capillaries, angiogenic gene expression and the angiogenic factors were increased after HCB-MNC injection. In the clinical study, the seven patients experienced no graft-versus-host-disease or cardiac/cerebral complications during the follow-up period. CONCLUSION: This preliminary study suggests that HCB-MNC might be a safe source of stem cells for treating ischaemic limbs. However, further clinical studies are needed to establish the long-term safety and the clinical efficacy of HCB-MNC transplantation in patients with ischaemic limbs.

Risk factors for device-associated infection related to organisational characteristics of intensive care units: findings from the Korean Nosocomial Infections Surveillance System.

Device-associated infections (DAIs) have been the major causes of morbidity and mortality of patients in intensive care units (ICUs). This study evaluated the risk factors for DAIs in ICUs. Ninety-six medical or surgical ICUs of 56 hospitals participated in the Korean Nosocomial Infections Surveillance System between July 2007 and June 2008. The occurrence of catheter-associated urinary tract infection (CAUTI), central line-associated bloodstream infection (CABSI), and ventilator-associated pneumonia (VAP) were monitored and DAI rates were calculated. Data associated with ICU characteristics were collected and Poisson regression was used for statistical analysis. Rates of CAUTI, CABSI, and VAP were 3.87 per 1000 urinary catheter days, 2.23 per 1000 central line days, and 1.89 per 1000 mechanical ventilator days, respectively. Rates of CAUTI were higher in ICUs in Seoul (P=0.032) and ICUs of major teaching hospitals (P=0.010). The ICUs of university-affiliated hospitals showed lower CAUTI rates (P=0.013). CABSI rates were higher in Seoul (P=0.001) and in medical ICUs (P=0.026). VAP rates were lower in ICUs of hospitals with more than 900 beds compared with hospitals with 400-699 beds (P=0.026). VAP rates were higher in surgical ICUs (P<0.0001) and increased 1.13-fold with each 100-unit increase in beds per infection control professional (P=0.003). The organisational and institutional characteristics of ICUs may influence DAI rates and there is a need for improvement in the incidence of VAP, CAUTI or CABSI.

TGF-beta type II receptor deficiency prevents renal injury via decrease in ERK activity in crescentic glomerulonephritis.

The role of transforming growth factor-beta (TGF-beta) receptor complex in the pathogenesis of crescentic glomerulonephritis (GN) is not clear. To test the hypothesis that TGF-beta signaling plays a crucial role in the development and progression of crescentic GN by inducing the activation of extracellular signal-regulated kinase (ERK) and expression of its target genes, anti-glomerular basement membrane (GBM) GN was induced in TGF-beta type II receptor (TGF-betaIIR) gene heterozygous (TGF-betaIIR(+/-)) C57BL/6J mice and wild-type animals. GN was initiated in preimmunized mice by administration of rabbit anti-mouse GBM serum. TGF-betaIIR deficiency was significantly associated with decreased renal damage at days 14, 21, and 28 after induction of GN: renal function impairment, proteinuria, proportion of crescents, glomerular accumulation of periodic acid-Schiff-positive material, relative cortical interstitial volume, as well as renal cortical phosphorylation of ERK and plasminogen activator inhibitor type I (PAI-1) and alpha2(I) collagen mRNA levels were significantly decreased in TGF-betaIIR(+/-) mice compared with wild-type animals. These results provide the first direct evidence that TGF-betaIIR deficiency protects against renal injury in crescentic GN, possibly by inhibiting the sustained activation of ERK and PAI-1 and alpha2(I) collagen gene expression. Thus, TGF-beta signaling appears to play an important role in the development and progression of crescentic GN by inducing the ERK activity, and PAI-1 and alpha2(I) mRNA expression.

Statistical evaluation of the cleft lip nose deformity image.

Cleft lip is a congenital deformity condition with separation of the two sides of the lip and causes nose deformity. Evaluation of surgical corrections and assessment of prognosis in nose deformity depend mainly on doctor's subjective judgment. Development of an objective assessment tool in evaluation of the cleft lip nose deformity patients will help in advancement and evaluation of surgical techniques. Therefore, our study aimed on quantitative assessment of a cleft lip nose deformity by comparing following parameters gathered from a photographic image of a cleft lip patient: (1) angle difference between two nostril axes, (2) center of the nostril and distance between two centers, (3) overlapped area of two nostrils and (4) the overlapped area ratio of two nostrils. Assessment results of the nose deformity were statistically analyzed with evaluation results from three highly experienced plastic surgeons. In addition, regression model was developed using correlation relationship and factor analysis of parameters from the results of image analysis.

Expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in human crescentic glomerulonephritis.

AIMS: In glomerulonephritis, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) may play important roles in the formation of crescents. These studies are designed to evaluate the expression patterns of ICAM-1 and VCAM-1 in human crescentic glomerulonephritis and to determine the cellular origin of adhesion molecules in the crescentic lesions. METHODS AND RESULTS: We examined the expression of ICAM-1 and VCAM-1 proteins in renal biopsies with cellular (n=7), fibrocellular (n=9) or fibrous (n=4) crescentic glomerulonephritis, and six controls by immunohistochemistry. mRNA expression of ICAM-1 and VCAM-1 was further evaluated by RNA in-situ hybridization. Cytokeratin or CD68 immunohistochemistry was performed on the same sections, where in-situ hybridization had been carried out. In cellular crescents, ICAM-1 and VCAM-1 proteins were over-expressed to a similar extent. Of the three types of crescents, the extent of ICAM-1 immunopositivity was the greatest in the cellular crescents and decreased towards the fibrous crescents (P < 0.05). Yet the extent of VCAM-1 immunoreactivity was not different between the types. Fibrous crescents still contained some epithelial cells and showed only VCAM-1 expression. In the glomeruli with cellular or fibrocellular crescents, the extent of ICAM-1 immunopositivity in the glomerular tufts was significantly larger than that of VCAM-1 (P < 0.05). In an in-situ hybridization study, the mRNA expression patterns of ICAM-1 and VCAM-1 paralleled their protein expressions. A double-labelling study showed that the signal for ICAM-1 and VCAM-1 mRNAs was mainly present in cytokeratin-positive and CD68-negative cells in the crescentic lesions. CONCLUSIONS: These results suggest that glomerular parietal epithelial cells in cellular crescents up-regulate both ICAM-1 and VCAM-1, and that some epithelial cells retained in fibrous crescents persistently over-express VCAM-1, but not ICAM-1. They also suggest that ICAM-1 is involved in early leucocyte recruitment into glomeruli in crescentic glomerulonephritis.

mRNA expression of urokinase and plasminogen activator inhibitor-1 in human crescentic glomerulonephritis.

AIMS: Weak staining for urokinase-plasminogen activator (uPA), tissue type plasminogen activator (tPA), or plasminogen activator inhibitor-1 (PAI-1) confined to crescents has been described in a few cases of severe crescentic glomerulonephritis. We evaluated the molecular mechanism by which these proteins are increased or induced within crescents. METHODS AND RESULTS: We examined uPA, tPA and PAI-1 mRNA expression in 12 renal biopsies with crescentic glomerulonephritis, and in six control renal biopsies with no detectable abnormalities by RNA in-situ hybridization. The expressions of uPA, tPA and PAI-1 proteins were also assessed by immunofluorescence. To better determine the cellular origin of uPA and PAI-1 transcripts, CD68 protein was studied by immunohistochemistry on the same sections on which in-situ hybridization had been performed. In controls, there were very low level signals of uPA and PAI-1 mRNAs in a few glomerular epithelial cells (GECs). Specific signals of uPA and PAI-1 mRNAs were detected in the cells forming crescents in all the cases with crescentic glomerulonephritis. However, weak expression of mRNA for tPA was detected in two cases only. Immunostaining for uPA and PAI-1 was positive in some but not all, cases of crescentic glomerulonephritis. A double-labelling study showed that the signal for PAI-1 and uPA mRNAs was mainly in CD68- cells. CONCLUSIONS: Local accumulation of uPA or PAI-1 in crescents is associated with enhanced mRNA expression of these proteins. The up-regulation of PAI-1 mRNA by GECs, in particular, could play a major role in the formation of persistent fibrin deposits and progression of the lesions in crescents. Whether up-regulation of uPA is an epiphenomenon or plays a pathogenic role in the formation of crescents remains to be clarified.

Advanced glycosylation end products stimulate collagen mRNA synthesis in mesangial cells mediated by protein kinase C and transforming growth factor-beta.

Advanced glycosylation end products (AGE) seem to be implicated in the pathogenesis of diabetic nephropathy. The present study has examined the effects of AGE on protein kinase C (PKC) activity and transforming growth factor-beta1 (TGF-beta1) in relation to collagen gene regulation in cultured human mesangial cells (HMCs). Quiescent HMCs were exposed to serum-free media containing bovine serum albumin (BSA), AGE-modified BSA (AGE-BSA), or glycated BSA in which AGE formation was prevented by the use of aminoguanidine (BSA-AM). AGE-BSA (200 microg/mL) induced a peak membrane-associated PKC activity, particularly PKC-a, at 4 hours. AGE-BSA stimulated alpha1(I) and alpha1(IV) collagen mRNA expression after 24-hour incubation with HMCs, which remained elevated until hour 60. HMCs incubated with AGE-BSA induced a significant inhibition of cell proliferation compared with cells incubated with BSA. AGE-BSA stimulated TGF-beta mRNA and protein expression in HMCs. The TGF-beta secreted by HMCs was shown by CCL-64 mink lung cell assay to be bioactive. In contrast, BSA-AM did not affect either collagen or TGF-beta mRNA or protein expression in HMCs. The stimulatory effects of AGE-BSA on collagen gene regulation in HMCs could be negated by the pretreatment of HMCs with GF 109203X for 30 minutes or with phorbol myristate acetate for 24 hours before AGE-BSA administration. Neutralizing antibody to TGF-beta inhibited increased collagen mRNA expression by HMCs exposed to AGE-BSA. These results suggest that AGE-BSA stimulates collagen mRNA expression by activating PKC and the transcriptional upregulation of TGF-beta1 in HMCs. Thus, PKC and TGF-beta may function as key signaling intermediaries in the AGE-up-regulated collagen gene expression pathway in HMCs.

The winged helix/forkhead transcription factor Foxq1 regulates differentiation of hair in satin mice.

Satin (sa) homozygous mice have a silky coat with high sheen arising from structurally abnormal medulla cells and defects in differentiation of the hair shaft. We demonstrate that the winged helix/forkhead transcription factor, Foxq1 (Forkhead box, subclass q, member 1) is mutant in sa mice. An intragenic deletion was identified in the radiation-induced satin mutant of the SB/Le inbred strain; a second allele, identified by an N-ethyl-N-nitrosourea (ENU) mutagenesis screen, has a missense mutation in the conserved winged helix DNA-binding domain. Homozygous mutants of the two alleles are indistinguishable. We show that Foxq1 is expressed during embryogenesis and exhibits a tissue-restricted expression pattern in adult tissues. The hair defects appear to be restricted to the inner structures of the hair; consequently, Foxq1 has a unique and distinct function involved in differentiation and development of the hair shaft. Despite an otherwise healthy appearance, satin mice have been reported to exhibit suppressed NK-cell function and alloimmune cytotoxic T-cell function. We show instead that the immune defects are attributable to genetic background differences.

mRNA expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in acute renal allograft rejection.

BACKGROUND: The intercellular adhesion molecule-1 (ICAM-1) and the vascular cell adhesion molecule-1 (VCAM-1) show a form of complementary distribution in normal and grafted kidneys. The molecular mechanism by which ICAM-1 and VCAM-1 are increased or induced on vascular cells during acute renal allograft rejection has not been clearly defined. METHODS: We examined ICAM-1 and VCAM-1 mRNA expression in 17 renal allograft biopsies with (n=12) and without (n=5) features of acute rejection, and four control renal biopsies with no detectable abnormalities by RNA in situ hybridization. The expression of ICAM-1 and VCAM-1 protein was also assessed by immunohistochemical staining of frozen sections. RESULTS: In controls and nonrejecting graft biopsies, the signals of the ICAM-1 and VCAM-1 transcripts in vascular cells were almost negligible. Specific signals of ICAM-1 and VCAM-1 mRNAs were detected on the endothelial cells of small muscular arteries in most cases with acute renal allograft rejection. The messages for ICAM-1 and VCAM-1 were also detected on arterial smooth muscle cells in all the five cases with severe type III rejection. CONCLUSIONS: These results suggest that the induced appearance of ICAM-1 and VCAM-1 on the vascular cells of acutely rejecting renal transplants was related to actual cellular synthesis and that both adhesion molecules could act together in the rejection process. They also suggest that the expression of ICAM-1 and VCAM-1 genes by arterial smooth muscle cells may be an important cause of transmural arteritis in severe acute renal allograft rejection.

High-level expression of canine parvovirus VP2 using Bombyx mori nucleopolyhedrovirus vector.

For the potential use as recombinant vaccine, canine parvovirus (CPV) major capsid protein VP2 was expressed using Bombyx mori nucleopolyhedrovirus (BmNPV) vector. CPV VP2 gene was introduced into polyhedrin-based BmNPV transfer vector pBmKSK3, and recombinant virus BmK1-Parvo was prepared. When anti-CPV.VP2 monoclonal antibody was employed in immunofluorescence staining, an intense signal was observed within BmK1-Parvo-infected Bm5 cells but not within uninfected cells or cells infected with a wild-type BmNPV-K1. In hemagglutination assay, the expression level of VP2 were 3.2 x 10(3) HA units/ml from infected Bm5 cells, 2.1x 10(5) HA units/larvae from infected larval fat body, and 1.6x 10(6) HA units/ml from infected larval hemolymph. These results suggested that BmNPV vector system using B. mori larva as host could be applied to efficient mass-production of recombinant vaccines.

Characterization of four isolates of Bombyx mori nucleopolyhedrovirus.

Bombyx mori nucleopolyhedroviruses (BmNPVs), isolated from a sericultural Korean farm, were purified and characterized by their DNA restriction pattern, virus replication, polyhedra production and gene structures. The EcoR I and Sal I fragments showed similar overall patterns with minor difference but distinguishable patterns in each isolate. There was no significant difference in the virus replication pattern, yield of total polyhedra production and polyhedra morphology, but the yield of released polyhedra by BmNPV-K1 in Bm5 cells was 2 to 5 times higher than that of other isolates. In comparative studies of p10 gene, BmNPV-K1 and K3 had same structure and they encoded a protein consisting of 94 amino acids. Although BmNPV-K2 encoded the same length of amino acids with BmNPV-K1 and K3, it had different structure, and BmNPV-K4 had the p10 gene encoding 70 amino acids.

mRNA expression of glomerular basement membrane proteins and TGF-beta1 in human membranous nephropathy.

Immunogold densities for the 'classical' and 'novel' alpha chains of type IV collagen, laminin, and fibronectin are increased in the spikes in human membranous nephropathy (MN). To investigate the molecular mechanisms which underlie these changes in glomerular basement membrane (GBM) components, alpha1(IV) collagen, alpha4(IV) collagen, S-laminin, fibronectin, transforming growth factor (TGF)-beta1 and TGF-beta2 mRNA expression was examined in 12 renal biopsy specimens with MN and six renal biopsies with no detectable abnormality by RNA in situ hybridization. In controls, there were relatively low signals of alpha1(IV) collagen, alpha4(IV) collagen, S-laminin, and TGF-beta1 mRNAs, but there were no fibronectin or TGF-beta2 transcripts in glomerular cells. In MN, the number of alpha4(IV) collagen, alpha1(IV) collagen, S-laminin or TGF-beta1 mRNA-expressing cells per glomerular cross-section was significantly larger than in controls (p< 0.05), and fibronectin mRNA was occasionally expressed in glomerular visceral epithelial cells (GECs). No message for TGF-beta2 was seen in MN. The number of TGF-beta1 mRNA-expressing cells per glomerular cross-section significantly correlated with that of alpha1(IV) mRNA-expressing cells (p< 0.01). The MN patients with positivie signal for fibronectin mRNA exhibited more severe GBM thickening than those without (p< 0.05). These results indicate that the increased presence of GBM proteins in spikes of MN is associated with enhanced mRNA expression of these proteins. They also suggest that subepithelial deposits in MN stimulate GECs to produce TGF-beta1, which in turn could mediate the expression of GBM protein genes by GECs.

LDL stimulates collagen mRNA synthesis in mesangial cells through induction of PKC and TGF-beta expression.

Abnormal lipid accumulation in glomeruli could be implicated in the pathogenesis of glomerulosclerosis. Low-density lipoprotein (LDL) stimulates collagen mRNA expression in cultured human mesangial cells (HMC). To explore the possible molecular mechanisms by which LDL promotes collagen gene expression, we examined the effects of LDL on protein kinase C (PKC) activity and transforming growth factor-beta (TGF-beta) expression in relation to collagen gene regulation in HMC. LDL (200 microg/ml) induced an acute increase in PKC activity, particularly PKC-alpha and -delta, within 15 min, which decreased to control value at 2 h. LDL stimulated TGF-beta1, and alpha1(I) and alpha1(IV) collagen mRNA expression within 30 min of incubation with HMC, and levels remained elevated until hour 4. LDL induced the secretion of TGF-beta by HMC. This TGF-beta was shown by CCL-64 mink lung cell assay to be, in part, bioactive. The stimulatory effects of LDL on collagen gene regulation in HMC were blocked by the inhibition of PKC using GF-109203X (GFX) or the downregulation of PKC using phorbol myristate acetate. Neutralizing antibody to TGF-beta inhibited the increased collagen mRNA expression by HMC exposed to LDL. The downregulation or inhibition of PKC did not affect the stimulatory effect of LDL on TGF-beta mRNA or protein expression. These results suggest that in HMC, LDL stimulates collagen mRNA expression through the rapid activation of PKC-alpha and -delta and transcriptional upregulation of TGF-beta. Thus PKC and TGF-beta may function as independent key signaling intermediaries in the pathway by which LDL upregulates collagen gene expression in HMC.

Pleiotropic skeletal and ocular phenotypes of the mouse mutation congenital hydrocephalus (ch/Mf1) arise from a winged helix/forkhead transcriptionfactor gene.

Congenital hydrocephalus is an etiologically diverse, poorly understood, but relatively common birth defect. Most human cases are sporadic with familial forms showing considerable phenotypic and etiologic heterogeneity. We have studied the autosomal recessive mouse mutation congenital hydrocephalus ( ch ) to identify candidate human hydrocephalus genes and their modifiers. ch mice have a congenital, lethal hydrocephalus in association with multiple developmental defects, notably skeletal defects, in tissues derived from the cephalic neural crest. We utilized positional cloning methods to map ch in the vicinity of D13Mit294 and confirm that the ch phenotype is caused by homozygosity for a nonsense mutation in a gene encoding a winged helix/forkhead transcription factor ( Mf1 ). Based on linked genetic markers, we performed detailed phenotypic characterization of mutant homozygotes and heterozygotes to demonstrate the pleiotropic effects of the mutant gene. Surprisingly, ch heterozygotes have the glaucoma-related distinct phenotype of multiple anterior segment defects resembling Axenfeld-Rieger anomaly. We also localized a second member of this gene family ( Hfh1 ), a candidate for other developmental defects, approximately 470 kb proximal to Mf1.

Infestation rate of head lice in primary school children in Inchon, Korea.

The survey of the head louse infestation of primary school children in Inchon city was carried out in May 26-27, 1995. Total number of the children surveyed were 1,530. Four (0.5%) out of 768 boys were infested with nits or adults/nymphs of lice and 72 (9.4%) out of 762 girls were positive with those of lice. The infestation rate of lice for girls was 19 times higher than that of boys.

[Seasonal prevalence and behaviour of Aedes togoi].

Ecological studies of Aedes togoi, the vector of malayan filariasis, were carried out at Tolsando, Yosu and Sokcho area in 1991. The adult population of Aedes togoi was continuously appeared from the first week of April to the end of November showing the highest density in July. The larvae of Aedes togoi were found at rock pools from March to December in Sokcho area and the density was highest in July and August, whereas in the southern coastal area (Yosu), the larvae were found throughout the year and the density was the highest in June. The rate of larvae inhabited below 0.5% salinity was 45.7% in Sokcho and 51.7% in Yosu. The feeding activity of Aedes togoi was nocturnal, with the peak period of 01:00-03:00 hours. Indoor feeding activities were slightly higher than outdoors showing the biting ratio of 1:0.8 (indoor: outdoor). The average number of Aedes togoi attracted to CO2 gas was 8.5 whereas 117 Anopheles sinensis was attracted. The result indicates that CO2 is not an effective attractant for host seeking of Aedes togoi compared to Anopheles sinensis. The most common place was bedroom with 54.5% of total collections and next to stock place (18.2%), floor (9.1%) and kitchen (9.1%).

[Effects of nutrient and salinity in egg and larval development of Aedes togoi].

The study made an observation on periodicity of oviposition, and the effects of nutrient and salinity in egg and larval development of Aedes togoi, and the results are summarized as follows: The 53.9% mosquitoes of one feeding laid eggs once, 26.9% laid twice and 19.2% laid three times. Autogenous rate of Aedes togoi reared in three different nutrient groups in larval sage was 6.9% in 0.8 mg/larva, 22.5% in 1.6 mg/larva and 44.4% in 2.4 mg/larva. The oviposition rate according to different salinity of the oviposition sites (0%, 0.5%, 1.0%, 2.0% and 4.0%) was 25.2% in distilled water, 36.2% in 0.5% salinity, 23.5% in 1.0% salinity and 14.6% in 2.0% salinity. Developing period of the aquatic stage of male Aedes togoi in 25 degrees C were shorter (10.73 days) than females (11.85 days). The most effective concentration of salinity for the developing period was 1.0% which took 9.25 days in males and 10.44 days in females. In the developmental status of the follicles according to nutrition in the larval stage, the numbers of follicles of groups fed 0.8 mg, 1.6 mg and 2.4 mg per larva were 180.7, 197.5 and 202 respectively. The result of ovary dissection on the 10th day after emergence, three different nutrition groups were in Christopher's stage IIa mostly; each 71.0%, 61.1% and 39.9% of the total follicles and autogenous females observed.

Clinical diversity and chromosomal localization of X-linked cone dystrophy (COD1).

X-linked progressive cone dystrophy (COD1) causes progressive deterioration of visual acuity, deepening of central scotomas, macular changes, and bull's-eye lesions. The cone electroretinography (ERG) is variably abnormal in affected males, and the rod ERG may also be abnormal. The clinical picture of heterozygous females ranges from asymptomatic to a widespread spectrum of cone-mediated dysfunction. A prior linkage study demonstrated linkage between the COD1 locus and the marker locus DXS84, assigned to Xp21.1, with no recombination. In the present study, we have clinically characterized a large four-generation family with COD1 and have performed a linkage analysis using seven polymorphic markers on the short arm of the X chromosome. No recombination was observed between the disease and the marker loci DXS7 and MAOA, suggesting that the location of COD1 is in the region Xp11.3, distal to DXS84 and proximal to ARAF1.