Glycosyl hydrolases of cell wall are induced by sugar starvation in Arabidopsis.

Three Arabidopsis genes encoding a putative beta-galactosidase (At5g56870), beta-xylosidase (At5g49360) and beta-glucosidase (At3g60140) are induced by sugar starvation. The deduced proteins belong to the glycosyl hydrolase families 35, 3 and 1, respectively. They are predicted to be secretory proteins that play roles in modification of cell wall polysaccharides based on amino acid similarity.

Concept and clinical application of cultivated epithelial transplantation for ocular surface disorders.

Corneal epithelial replacement using a tissue engineering technique holds much promise for ocular surface reconstruction in cases of corneal epithelial stem cell deficiency. However, even though an autologous cultivated corneal epithelial stem cell sheet is the safest and most reliable form of sheet, bilaterally affected ocular surface disorders cannot be treated by this method. To treat bilateral cases, we must choose either an allogeneic cultivated corneal epithelial sheet or an autologous cultivated oral mucosal epithelial sheet.

A comparison between cultivated and conventional limbal stem cell transplantation for Stevens-Johnson syndrome.

Purpose: To compare the resolution of inflammation and long-term results of cultivated and conventional limbal stem cell transplantation (LSCT) in a patient with Stevens-Johnson syndrome (SJS).

Design: Interventional case report.

Methods: A 32-year-old man with SJS and bilateral total limbal stem cell deficiency underwent cultivated LSCT in the right eye, followed by conventional LSCT in the left eye three weeks later.

Autologous serum-derived cultivated oral epithelial transplants for severe ocular surface disease.

Objective: To evaluate the use of autologous serum (AS)-derived cultivated oral epithelial transplants for the treatment of severe ocular surface disease.

Methods: We used AS from 10 patients with severe ocular surface disease and total limbal stem cell deficiency to develop autologous cultivated oral epithelial equivalents. These were compared with epithelial equivalents derived from conventional fetal bovine serum-supplemented medium.

Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon.

Adenovirus (Ad) vectors are widely used in gene therapy and in vitro/in vivo gene transfer because of their high transduction efficiency. However, Ad vector application in the gene therapy field is limited by poor transduction into cells not expressing the primary receptor, coxsackievirus and adenovirus receptor. To overcome this problem, several types of capsid-modified Ad vectors have been developed.

Ocular surface reconstruction with combination of cultivated autologous oral mucosal epithelial transplantation and penetrating keratoplasty.

Purpose: To report an assessment of the two-step surgical combination of cultivated autologous oral mucosal epithelial transplantation (COMET) and penetrating keratoplasty (PKP) used to treat patients with severe limbal deficiency disorders, and to investigate the keratin expression patterns of transplanted surviving oral mucosal epithelium.

Design: Observational case series.

Methods: Two patients with Stevens-Johnson syndrome and chemical eye injury were treated by COMET followed, approximately six months later, by a PKP triple procedure.

Osteolipoma arising adjacent to the sternoclavicular joint. A case report.

A 45-year-old woman presented with one-year history of a mass on her chest. Computed tomography and magnetic resonance imaging demonstrated a tumour near the sternoclavicular joint. The tumour was diagnosed as osteolipoma histologically after resection.

Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation.

Purpose: The corneal epithelium is essential for maintaining corneal transparency, and efforts have been made to develop improved techniques for corneal epithelial transplantation in patients with total limbal failure. We evaluated the suitability of transplanted cultivated human conjunctival epithelium (HCjE) as a corneal epithelium replacement in rabbits with total corneal and limbal deficiency.

Methods: HCjE cells, cultivated on human amniotic membrane (AM) to confluence and exposed to an air-liquid interface (air-lifted), were transplanted onto denuded rabbit corneas and monitored for 2 weeks.

Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease.

Purpose: To evaluate the use of autologous serum (AS)-derived cultivated corneal epithelial transplantation for the treatment of severe ocular surface disease.

Design: Retrospective noncomparative case series.

Participants: Nine eyes from 9 patients with total limbal stem cell deficiency were studied.

Molecular characterization of lipoprotein lipase, hepatic lipase and pancreatic lipase genes: effects of fasting and refeeding on their gene expression in red sea bream Pagrus major.

To investigate the nutritional regulation of lipid metabolism in fish, molecular characterization of lipases was conducted in red sea bream Pagrus major, and the effects of fasting and refeeding on their gene expression was examined. Together with data from a previous study, a total of four lipase genes were identified and characterized as lipoprotein lipase (LPL), hepatic lipase (HL) and pancreatic lipase (PL). These four lipase genes, termed LPL1, LPL2, HL and PL, share a high degree of similarity.

The GrlR-GrlA regulatory system coordinately controls the expression of flagellar and LEE-encoded type III protein secretion systems in enterohemorrhagic Escherichia coli.

The gene function of the locus of enterocyte effacement (LEE) is essential for full virulence of enterohemorrhagic Escherichia coli (EHEC). Strict control of LEE gene expression is mediated by the coordinated activities of several regulatory elements. We previously reported that the ClpX/ClpP protease positively controls LEE expression by down-regulating intracellular levels of GrlR, a negative regulator of LEE gene expression.

Leptospirosis in squirrels imported from United States to Japan.

We diagnosed leptospirosis in 2 patients exposed to southern flying squirrels imported from the United States to Japan. Patients worked with exotic animals in their company. Leptospira isolates from 1 patient and 5 of 10 squirrels at the company were genetically and serologically identical and were identified as Leptospira kirschneri.

Induction of BiP by sugar independent of a cis-element for the unfolded protein response in Arabidopsis thaliana.

BiP is a molecular chaperone induced in the unfolded protein response (UPR). In mammalian cells, BiP is induced by glucose starvation when it is called glucose-regulated protein 78 (GRP78). In Arabidopsis thaliana, however, we demonstrated that BiP transcripts decreased with sugar depletion and increased with sugar addition.

High volume hydrodynamic injection of plasmid DNA via the hepatic artery results in a high level of gene expression in rat hepatocellular carcinoma induced by diethylnitrosamine.

Background: Hydrodynamic injection of naked plasmid DNA (pDNA) via the tail vein is a safe and effective method of gene transfer to the liver. However, successful gene transfer has yet to be shown for hepatocellular carcinoma (HCC); therefore, we investigated the feasibility and efficacy of hydrodynamic injection via the tail vein and hepatic artery in a diethylnitrosamine (DEN)-induced HCC model in rats.

Methods: HCC was induced in Sprague-Dawley rats by 100 ppm DEN in drinking water.

Efficient gene transfer into differentiated human trophoblast cells with adenovirus vector containing RGD motif in the fiber protein.

Previously we reported fiber-modified adenovirus (Ad) vectors containing the Arg-Gly-Asp (RGD) motif on the HI loop of the fiber knob (Ad-RGD vectors) have high gene transfer efficacy into some human trophoblast cell lines. In the current study, we investigate transgene activity of Ad-RGD during differentiation of human cytotrophoblast BeWo cells into syncytiotrophoblast-like cells. Although cellular differentiation into syncytiotrophoblast cells was followed by a decrease in the coxsackievirus and adenovirus receptor levels on the cell membrane, the alphaVbeta3 and alphaVbeta5 integrin levels did not change.

Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells.

Background: We have previously developed a limbal epithelial culture system using a cell-suspension method on denuded amniotic membrane (AM). However, other workers reported that intact AM is advantageous for limbal epithelial culture in that it preserves stem cell characteristics. In this study, we cultivated human limbal epithelial cell-suspensions on both intact and denuded AM and compared the morphology and adhesion of the limbal epithelial cells on these two substrates.

Molecular identification of a novel beta-1,3-glucanase from alkaliphilic Nocardiopsis sp. strain F96.

Alkaliphilic Nocardiopsis sp. strain F96 produced three beta-1,3-glucanase isozymes of different molecular masses (BglF1, BglF2 and BglF3). The N-terminal amino acid sequences of BglFs indicated that these isozymes were the products of a single gene.

Adenovirus vector-mediated gene transfer into stem cells.

Stem cells, including embryonic stem (ES) cells, mesenchymal stem cells (MSCs), and hematopoietic stem cells (HSCs), are defined by their capacity for self-renewal and multilineage differentiation. Efficient gene transfer into stem cells is essential for the basic research in developmental biology and for therapeutic applications in gene-modified regenerative medicine. Adenovirus (Ad) vectors, based on Ad type 5, can efficiently and transiently introduce the exogenous gene into many cell types via the primary receptor, coxsackievirus, and adenovirus receptor (CAR).

Modified adenoviral vectors ablated for coxsackievirus-adenovirus receptor, alphav integrin, and heparan sulfate binding reduce in vivo tissue transduction and toxicity.

Coxsackievirus and adenovirus receptor (CAR), alphav integrins, and heparan sulfate glycosaminoglycans (HSGs) are the tropism determinants of adenoviral (Ad) vectors in vivo. For the development of a targeted Ad vector, its broad tropism needs to be blocked (or reduced). We have previously developed Ad vectors with ablation of CAR, alphav integrin, and HSG binding by mutation of the FG loop in the fiber knob (deletion of T489, A490, Y491, and T492 of the fiber protein), deletion of the RGD motif of the penton base, and substitution of the fiber shaft domain for that derived from Ad type 35, respectively, and have shown that this triple-mutant Ad vector [Ad/deltaF(FG)deltaP-S35-L2] exhibits significantly lower transduction in mouse liver compared with the conventional Ad vector [Koizumi, N.

Adenovirus serotype 35 vector-mediated transduction into human CD46-transgenic mice.

We previously demonstrated that systemic administration of adenovirus serotype 35 (Ad35) vectors to mice does not mediate efficient transduction in organs, probably because expression of the mouse analog of the subgroup B Ad receptor, human CD46 (membrane cofactor protein), is limited to the testis. Here, we describe the in vitro and in vivo transduction characteristics of Ad35 vectors by using homozygous and hemizygous human CD46-transgenic (CD46TG) mice, which ubiquitously express human CD46. An Ad35 vector more efficiently transduced the primary dendritic cells and macrophages prepared from CD46TG mice than those from wild-type mice.

Cytomegalovirus in aqueous humor from an eye with corneal endotheliitis.

Purpose: To report cytomegalovirus (CMV) DNA in aqueous humor from a patient with unilateral corneal endotheliitis.

Design: Case report.

Methods: A 51-year-old man presented with unilateral corneal endotheliitis with linear keratic precipitates and coin-shaped lesions.

Primary anetoderma: a case report and its modified classification.

A fifty-nine-year-old healthy male presented with fifteen round pouches around his bilateral shoulders and proximal thighs, at which elasticity was lost on palpation. Histopathological examinations demonstrated that the lesional dermis was thinner than normal skin and there was loss of elastic fibers and mild inflammatory cell infiltration. Because there was no preceding inflammatory skin lesion or associated diseases, the patient was diagnosed with primary anetoderma.

Sporadic cases and an outbreak of leptospirosis probably associated with recreational activities in rivers in the northern part of Okinawa Main Island.

In the summer of 2003, sporadic cases and an outbreak of human leptospirosis probably related to recreation in rivers occurred in the northern part of Okinawa Main Island. Sixteen of 22 suspected cases were definitely diagnosed as leptospirosis by serological test or isolation. The infective leptospiral serovar in 14 cases was presumed to be Hebdomadis.

Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation.

Purpose: To perform a midterm assessment of the integrity and reproducibility of cultivated autologous oral mucosal epithelial sheets, and to evaluate the clinical efficacy of their transplantation in ocular surface.

Design: Observational case series.

Methods: Cultivated autologous oral mucosal epithelial sheets were created using amniotic membrane and buccal mucosal epithelium from 12 patients with Stevens-Johnson syndrome, chemical and thermal injury, pseudo-ocular cicatricial pemphigoid, and idiopathic ocular surface disorder.