Staphylococcus aureus-derived membrane vesicles exacerbate skin inflammation in atopic dermatitis.

BACKGROUND: Skin colonization or infection with Staphylococcus aureus is known to trigger aggravation of atopic dermatitis (AD). However, the exact mechanisms by which S. aureus can worsen AD are unknown. OBJECTIVE: We investigated whether and how S. aureus-derived membrane vesicles (MVs) contribute to worsening of AD. METHODS: Immunohistochemical and immunoelectron microscopic analyses were performed to detect staphylococcal protein A (SPA) in the epidermis of AD lesions. HaCaT cells were treated with S. aureus MVs and were analysed for the expression of cytokine genes. Immunopathology and cytokine gene profiles were analysed after topical application of S. aureus MVs to AD-like skin lesions in a mouse model. RESULTS: The MV component SPA was detected in the keratinocytes as well as in the intercellular space of the epidermis of AD lesions colonized with S. aureus. Intact MVs from S. aureus delivered their components to keratinocytes and stimulated pro-inflammatory cytokine gene expression in vitro. A knock-down of Toll-like receptor 2 or nucleotide-binding oligomerization domain 2 using small interfering RNAs suppressed interleukin-8 gene expression. Topical application of intact S. aureus MVs to AD-like skin lesions in the mouse model induced massive infiltration of inflammatory cells and the resulting eczematous dermatitis. This inflammatory reaction was associated with a mixed Th1/Th2 immune response and enhanced expression of chemokine genes in AD-like skin lesions. CONCLUSIONS AND CLINICAL RELEVANCE: This study showed the importance of S. aureus MVs as a potent mediator for worsening of AD among many exogenous worsening factors of AD. Thus, S. aureus MVs may be regarded as one of the therapeutic targets for the management of AD aggravation.

PATHOME: an algorithm for accurately detecting differentially expressed subpathways.

The translation of high-throughput gene expression data into biologically meaningful information remains a bottleneck. We developed a novel computational algorithm, PATHOME, for detecting differentially expressed biological pathways. This algorithm employs straightforward statistical tests to evaluate the significance of differential expression patterns along subpathways. Applying it to gene expression data sets of gastric cancer (GC), we compared its performance with those of other leading programs. Based on a literature-driven reference set, PATHOME showed greater consistency in identifying known cancer-related pathways. For the WNT pathway uniquely identified by PATHOME, we validated its involvement in gastric carcinogenesis through experimental perturbation of both cell lines and animal models. We identified HNF4α-WNT5A regulation in the cross-talk between the AMPK metabolic pathway and the WNT signaling pathway, and further identified WNT5A as a potential therapeutic target for GC. We have demonstrated PATHOME to be a powerful tool, with improved sensitivity for identifying disease-related dysregulated pathways.

Enzymatic cleavage of fusion proteins using immobilised protease 3C.

A strategy for efficient cleavage of fusion proteins using an immobilised protease has been developed. Protease 3C from coxsackie virus was recombinantly produced in Escherichia coli and covalently immobilised onto a solid support. Thereafter, Z(basic) tagged fusion proteins, with a specific cleavage sequence between the domains, were flown through the proteolytic column and circulated until complete cleavage. Subsequently, the processed protein solution was applied on a cation exchanger. Thereby, removal of the released, positively charged fusion tag, Z(basic), was done by adsorption to the matrix while the target proteins were recovered in the flow through. Interestingly, the columns were shown to be reusable without any measurable decrease in activity. Moreover, after storage in 4 degrees C for two months the activity was almost unaffected.

Stable transformation of rice (Oryza sativa L.) via microprojectile bombardment of highly regenerative, green tissues derived from mature seed.

A highly efficient and reproducible transformation system for rice ( Oryza sativa L. cv. Taipei 309) was developed using microprojectile bombardment of highly regenerative, green tissues. These tissues were induced from mature seeds on NB-based medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP) and high concentrations of cupric sulfate under dim light conditions; germinating shoots and roots were completely removed. Highly regenerative, green tissues were proliferated on the same medium and used as transformation targets. From 431 explants bombarded with transgenes [i.e. a hygromycin phosphotransferase ( hpt) gene plus one of a wheat thioredoxin h ( wtrxh), a barley NADP-thioredoxin reductase ( bntr), a maize Mutator transposable element ( mudrB) or beta-glucuronidase ( uidA; gus) gene], 28 independent transgenic events were obtained after an 8- to 12-week selection period, giving a 6.5% transformation frequency. Of the 28 independent events, 17 (61%) were regenerable. Co-transformation of the second introduced transgene was detected in 81% of the transgenic lines tested. Stable integration and expression of the foreign genes in T(0) plants and T(1) progeny were confirmed by DNA hybridization, western blot analyses and germination tests.

Comparison of visual field defects between primary open-angle glaucoma and chronic primary angle-closure glaucoma in the early or moderate stage of the disease.

The visual field data (Humphrey C24-2) of early to moderate stage glaucoma between primary open-angle glaucoma (POAG) and chronic primary angle-closure glaucoma (PACG) patients was compared. Eleven POAG (11 eyes) and 14 chronic PACG patients (14 eyes) were matched with respect to age, sex, and disease severity. Although the mean deviation, pattern standard deviation, and short-term fluctuation were similar between the two groups, the mean corrected pattern standard deviation was higher in the POAG patients (p=0.048). When the numeric pattern deviation data (STATPAC II) was used for point-wise between-group comparisons, more depressed paracentral points were found in POAG patients. Different patterns were identified in visual field damages between the chronic PACG and POAG patients. The chronic PACG patients showed more generalized field loss than the POAG patients.

Effect of cataract extraction on blue-on-yellow visual field.

PURPOSE: It is assumed that cataract can influence the results of blue-on-yellow perimetry. However, actual sensitivity change in blue-on-yellow perimetry before and after cataract surgery has not been fully demonstrated. METHODS: Prospective. SETTING: Institutional. STUDY POPULATION: Twenty-two eyes of 22 consecutive patients without ocular pathology, other than cataract, known to influence visual field undergoing cataract surgery. OBSERVATION PROCEDURES: Both white-on-white (W-W) and blue-on-yellow (B-Y) perimetries. MAIN OUTCOME MEASURES: Global indexes including mean deviation and other pertinent data of white-on-white and blue-on-yellow perimetries were compared before and after cataract surgery. RESULTS: Mean deviation was improved after cataract surgery in both white-on-white perimetry (from -6.88 dB to -3.36 dB, P <.0001) and blue-on-yellow perimetry (from -12.22 dB to -3.64 dB, P <.0001, paired t test). However, the mean difference between preoperative and postoperative mean deviation in blue-on-yellow perimetry (8.58 +/- 3.96 dB) was significantly higher than that of white-on-white perimetry (3.52 +/- 2.69 dB; P <.0001, unpaired t test). That is, the change in mean deviation was greater in blue-on-yellow perimetry than in white-on-white perimetry by a factor of 2.4 times. CONCLUSION: Cataract causes predominantly a general reduction of sensitivity in both blue-on-yellow and white-on-white perimetries, and the general reduction of blue-on-yellow sensitivity is far greater than that of white-on-white sensitivity.

Expression of mucosal cyto-chemokine mRNAs in patients with Helicobacter pylori infection.

BACKGROUND: Helicobacter pylori-induced destruction of the gastroduodenal mucosal barrier is initiated with mucosal infiltration of inflammatory cells. Cytokines and chemokines have been suggested to play important roles in the migration and activation of these inflammatory cells into the mucosa. The present study aimed to investigate expression rates of cyto-chemokine mRNAs using gastric mucosal biopsy specimens. METHODS: In 98 patients infected with Helicobacter pylori, mucosal mRNA expression rates of cytokines (IL-1 beta, IL-6, and IL-10), C-C chemokines (macrophage inflammatory protein 1 alpha (MIP-1 alpha), and macrophage inflammatory protein 1 beta (MIP-1 beta), monocyte chemotactic and activating factor (MCAF), regulated on activation, normal T cell expressed and presumably secreted (RANTES)) and C-X-C chemokines (IL-8 and growth regulated a (GRO-alpha)) were examined using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The expression rates of mRNA for IL-8, GRO-alpha, MIP-1 alpha and RANTES were significantly more increased in H. pylori-positive patients than in H. pylori-negative patients. However, the expressions of IL-1 beta, IL-6 and IL-10 mRNA were statistically not different between two groups. After eradication of H. pylori, expressions of mRNA for three cytokines (IL-1 beta, IL-6 and IL-10), four C-C chemokines (MIP-1 alpha, MIP-1 beta, MCAF and RANTES) and two C-X-C chemokines (IL-8 and GRO-alpha) were significantly decreased. CONCLUSION: These results suggest that C-X-C chemokines and some C-C chemokines play important roles in H. pylori-associated peptic ulcer diseases.

Metabolic adaptations to dietary fat malabsorption in chylomicron-deficient mice.

A mouse model of chylomicron deficiency was recently developed; these mice express a human apolipoprotein (apo) B transgene in the liver but do not synthesize any apoB in the intestine. Despite severe intestinal fat malabsorption, the mice maintain normal concentrations of plasma lipids and liver-derived apoB 100-containing lipoproteins. We investigated the metabolic mechanisms by which plasma lipid levels are kept normal. De novo lipogenesis (DNL) and cholesterogenesis were measured by mass isotopomer distribution analysis (MIDA). Plasma non-esterified fatty acid (NEFA) fluxes and hepatic re-esterification of labelled plasma NEFA were also measured. Hepatic and plasma triacylglycerol (TG) concentrations and plasma NEFA fluxes were not different between chylomicron-deficient mice and controls. The contribution from DNL to the hepatic TG pool was only modestly higher in chylomicron-deficient mice [12+/-2.1% (n=7) compared with 3.7+/-1.0% (n=9); means+/-S.E.M.], whereas cholesterogenesis was markedly elevated. The fractional contribution from plasma NEFA to hepatic TG was greatly elevated in the chylomicron-deficient animals (62% compared with 23%). Accordingly, 73% of hepatic TG was neither from DNL nor from plasma NEFA in controls, presumably reflecting prior contribution from chylomicron remnants, compared with only 26% in the chylomicron-deficient group. The long-term contribution from DNL to adipose fat stores reached approximately the same steady-state values (approximately 30%) in the two groups. Body fat accumulation was much lower in chylomicron-deficient animals; thus, whole-body absolute DNL was significantly lower. We conclude that plasma and hepatic TG pools and hepatic secretion of apoB-containing particles are maintained at normal levels in chylomicron-deficient mice, not by de novo fatty acid synthesis, but by more avid re-esterification of plasma NEFA, replacing the normally predominant contribution from chylomicrons, and that some dietary fat can be absorbed by apoB-independent mechanisms.

Streptococcal keratitis after myopic laser in situ keratomileusis.

A 24-year-old healthy male underwent uncomplicated laser in situ keratomileusis (LASIK) in left eye. One day after the surgery, he complained of ocular pain and multiple corneal stromal infiltrates had developed in left eye. Immediately, the corneal interface and stromal bed were cleared, and maximal antibiotic treatments with fortified tobramycin (1.2%) and cefazolin (5%) were given topically. The causative organism was identified as 'Streptococcus viridans' both on smear and culture. Two days after antibiotic therapy was initiated, the ocular inflammation and corneal infiltrates had regressed and ocular pain was relieved. One month later, the patient's best corrected visual acuity had returned to 20/20 with -0.75 -1.00 x 10 degrees, however minimal stromal scarring still remained. This case demonstrates that microbial keratitis after LASIK, if treated promptly, does not lead to a permanent reduction in visual acuity.

Clarifying the nomenclature for primary angle-closure glaucoma.

Despite tremendous development in the field of glaucoma, there is confusion in the nomenclature system for classification, gonioscopy, and mechanisms of primary angle-closure glaucoma. In this article, we critically review the various nomenclature systems and suggest some modifications to the current classifications. These include four diagnostic categories: 1) angle-closure glaucoma suspect; 2) angle-closure hypertension; 3) chronic angle-closure glaucoma; and 4) acute angle-closure glaucoma. Gonioscopy is valuable for evaluating primary angle-closure glaucoma but more precise descriptions of gonioscopic observations are needed. Because of racial differences in the prevalence and mechanisms of primary angle-closure glaucoma, worldwide population-based study is necessary. Newer technologies, such as ultrasound biomicroscopy, show promise for the study of primary angle-closure glaucoma.

Dilated, miotic-resistant pupil and laser iridotomy in primary angle-closure glaucoma.

We analyzed 22 eyes with primary angle-closure glaucoma that underwent initial laser iridotomy to determine which factors could lead to subsequent trabeculectomy. Twenty-two eyes were divided into two groups: (1) the eyes in which intraocular pressure (IOP) could be controlled by iridotomy and/or topical medication (iridotomy success group, 15 eyes) and (2) the eyes that underwent trabeculectomy to control IOP in spite of a patent opening (iridotomy failure group, 7 eyes). The clinical variables between the two groups were analyzed. Age, sex, visual field defect, presenting IOP and cup/disk ratio were not significantly different between the iridotomy success and failure groups. However, presence of peripheral anterior synechiae (PAS) greater than 50% was found more frequently in the iridotomy failure group as compared with the iridotomy success group (4/7 vs. 1/15, p = 0.02). Dilated, miotic-resistant pupils were observed only in the iridotomy failure group (4/7 vs. 0/15, p = 0.004). PAS greater than 50% and dilated, fixed pupils were observed in these same cases (4 eyes). Our results suggest that laser iridotomy may not be helpful in cases with dilated and miotic-resistant pupils with formation of extensive PAS.

[Recent perceptions about chromosome pairing in meiosis]. / Neue Erkenntnisse über die Chromosomenpaarung in der Meiose.

Recent results from meiotic research in animals are presented with emphasis on the pairing behaviour of autosomes und sex chromosomes. Synaptonemal complex preparations were primarily utilized, that allowed observations of the pairing behaviour of the sex chromosomes is quite remarkable. The structure and function of the synaptonemal complex is discussed and the localization of the pseudo autosomal region on the sex chromosomes is described.

Laser assisted in situ keratomileusis for high myopia.

BACKGROUND AND OBJECTIVE: We present the clinical results of laser assisted in situ keratomileusis (LASIK) to correct myopia of more than -10.00 diopters (D). PATIENTS AND METHODS: Eighteen eyes are included in this study. Pre-operative spherical equivalent was -16.40 D, range from -10.50 to -21.50 D. Hinge corneal cap techniques were used with automated microkeratome (Steinway, USA). The thickness of the flap was 130 microns. The stromal bed was ablated with the OmniMed II excimer laser, using the standard MKM program (Summit Technology, Inc., Waltham, MA). The ablation zone was between 4.2 and 5.0 mm. RESULTS: At 6 months postoperatively, the mean spherical equivalent was +0.14 D with a range of +3.50 to -2.00 D. A result within 2.00 D of the desired correction was achieved in 93% of eyes, while a result within 1.00 D was achieved in 47%. Uncorrected visual acuity was improved in all cases. One eye lost greater than two lines of pre-operative best corrected vision due to macular hemorrhage. CONCLUSION: These data show that the LASIK procedure appears to be effective for correcting high myopia.

The effect of flat anterior chamber on the success of trabeculectomy.

The authors reviewed 70 eyes of 59 patients who underwent trabeculectomy. We divided the eyes into the flat anterior chamber group (23 eyes) and formed anterior chamber group (47 eyes, control group). The influence of clinical variables on the development of flat anterior chamber, and the effect of flat anterior chamber on the success of trabeculectomy were evaluated. The two groups were similar in terms of many variables, but the preoperative intraocular pressure (27.1 +/- 15.4 vs. 18.1 +/- 8.7 mmHg), incidence of cataract progression (9/18 vs. 10/44) and choroidal detachment (5/23 vs. 2/47) were higher in the flat anterior chamber group (p < 0.05). The overall success rates for the follow-up period of 30 months were similar between the two groups (p > 0.05). However, the short-term (2 to 8 months) postoperative success rates were lower in the flat anterior chamber group (p < 0.05). The development of flat anterior chamber following trabeculectomy can be reduced by lowering the preoperative intraocular pressure, and flat anterior chamber may be harmful to the postoperative course of trabeculectomy.

Multizone photorefractive keratectomy for myopia of 9 to 14 diopters.

We performed photorefractive keratectomy (PRK) using new multizone software of the Summit Omnimed excimer laser (Summit Technology, Inc., Waltham, Mass.) on thirteen highly myopic eyes. Average preoperative myopia was -10.94 diopters (D) (range -8.60 D to -13.50 D). Mean postoperative spherical equivalent refraction was -0.15 D (range +1.50 D to -4.00 D), at 3 months postoperatively. Uncorrected visual acuity of all the eyes improved markedly. There was no serious complication except one eye that lost two lines of spectacle corrected visual acuity possibly due to decentration. Though we need longer follow-up data and a larger sample size, the new multizone software seems to be effective for correcting high myopia.

A balanced autosomal reciprocal translocation in an azoospermic bull.

This appears to be the first reported case of a bull with a balanced autosomal reciprocal translocation associated with azoospermia. The analysis includes somatic chromosome banding, conventional meiotic analysis, and electron microscopy of synaptonemal complexes (SCs). The karyotype of the bull was found to be 60,XY,rcp(8;13)(q11;q24). Electron microscopy of SCs in microspread pachytene spermatocytes revealed a high incidence of terminal asynapsis of the smallest arm of the quadrivalent. Most quadrivalents with such asynapsis and few with nonhomologous synapsis showed associations with the XY sex bivalent, leading to complete meiotic arrest at late pachynema. Except for one diakinetic cell, no diplotene and subsequent stages were encountered in air-dried meiotic preparations. The presence of degenerating primary spermatocytes in SC preparations, as well as in testicular sections, and the absence of spermatozoa in ejaculates confirm the chromosomally derived male sterility of the bull. X-chromosome reactivation, evidenced by the cytomorphological reversal of associated sex bivalents, appeared to be the initial step in the degeneration of spermatocytes. Consequently, the formation of a separate, fully developed XY body, which was previously demonstrated on the periphery of spermatocyte nuclei in fertile bulls, could not be attained in this case. Extensive end-to-end association of autosomal bivalents in meiotically arrested, as well as degenerating, spermatocytes was a consistent and unique observation of this study. Such associations may lead to enhanced reactivation of the X chromosome.

A reciprocal whole-arm translocation, rcp(1;6)(1p6p;1q6q) in a boar, localization of the breakpoints, and reassignment of the genes for glucose phosphate isomerase (GPI) and calcium release channel (CRC).

A reciprocal whole-arm translocation between chromosomes 1 and 6 in a Swiss Large White boar with reduced fertility was identified by the use of different staining techniques in mitotic metaphase cells, synaptonemal complex analyses, and meiotic chromosome preparations. The karyotype of this boar was demonstrated to be 38,XY,rcp(1;6)(1p6p;1q6q). To further localize the breakpoints more precisely and determine the precise gene locations, several in situ hybridization experiments were performed with a chromosome 1 centromere-specific probe and two other gene probes. The breakage and reunion points of both chromosomes were located in the centromeric regions. The genes for glucose phosphate isomerase and calcium release channel were mapped to 6cen----q12.