Morphometric analyses in patients treated with subthreshold laser photocoagulation for central serous chorioretinopathy.

BACKGROUND AND OBJECTIVE: To analyze changes in selected parameters in optical coherence tomography (OCT) after subthreshold laser coagulation (ST-LP) in patients with central serous chorioretinopathy (CSCR). MATERIALS AND METHODS: Fifty-four eyes of 49 patients with CSCR were included in the study. Each patient underwent therapy with ST-LP with a frequency-doubled Neodym-YAG Laser and OCT imaging. In OCT the thickness of the central subfield, cube volume, average cube thickness, volume under the ETDRS grid, and average thickness under the ETDRS grid were collected. RESULTS: Decreases in total OCT volume and central retinal subfield thickness were statistically significant (p < 0.05). Possible correlations were observed between visual acuity at V3 (3 months after ST-LP) and Baseline and between central retinal subfield thickness at V1 (4 weeks after ST-LP) and visual acuity at BL. CONCLUSION: A decrease in retinal thickness and retinal volume could be shown after ST-LP. Central retinal subfield thickness measured by OCT could be a more sensitive measure than mean retinal thickness or macular volume for early detection of disease recurrence occurring in some patients 3 months after ST-LP.

[New Possibilities in Retinal Diagnostics Using OCT Angiography]. / Neue Möglichkeiten in der retinalen Diagnostik mittels OCT-Angiografie.

BACKGROUND: Instruments for using OCT angiography (OCTA) in daily clinical practice have recently become available. The aim of this paper is to report the possibilities, advantages and limitations of OCTA in the clinical diagnosis of diseases of the posterior segment of the eye. PATIENTS/METHODS: Patients with diabetic retinopathy, retinal vascular occlusions, and age-related macular degeneration who had been assigned to fluorescein angiography (FA) additionally underwent an AngioPlex™-OCTA examination, which captures a 6 × 6 mm scanning area centred on the fovea. If deemed necessary, 3 × 3 mm volume scans were created in regions of interest. The findings of FA and OCTA were correlated and compared. RESULTS: The OCTA procedure took only a few seconds, was easily integrated into the standard OCT diagnostic imaging procedure, and delivered highly detailed, three dimensional images of the entire microvasculature of the retina and choroid. Microvascular changes, such as microaneurysms, intraretinal microvascular abnormalities, non-perfused areas, alterations in the foveal avascular zone (FAZ) and neovascularization were reliably detected. Overall, OCTA results were in good agreement with the results of the FA. Additionally, OCTA provided more detailed and three dimensional information and thus permitted a better assessment of the spatial extension of microvascular abnormalities. Due to OCTA's limited scanning area, vascular alterations in the mid-periphery were detected only if their location had already been determined by FA. Although OCTA does not show leakage, macular oedema can be diagnosed from OCTA, together with OCT thickness measurements. CONCLUSION: OCTA provides important three dimensional information on vascular alterations and is already an indispensable diagnostic method. As the procedure takes just a few seconds and can be performed non-invasively, OCTA is well suited as a monitoring method and may allow early diagnosis. In this sense, prospective studies are required to determine precise OCTA analytical strategies for specific diseases. It is very likely that OCTA will revolutionise the diagnosis of retinal and choroidal diseases; however, it is not yet clear estimated to what extent it will replace FA.

Effect of E. coli Nissle 1917 on post-inflammatory visceral sensory function in a rat model.

OBJECTIVE: Visceral hyperalgesia (VH) plays a key role for the manifestation of functional gastrointestinal (GI) disorders. In a subgroup of patients, the initial manifestation is preceded by GI inflammation. Recent studies have demonstrated an improvement of inflammation and symptoms during treatment with Escherichia coli Nissle 1917 (EcN). AIM: We aimed to characterize the effects of EcN on visceral sensitivity in a rat model of post-inflammatory VH. METHODS: Male Lewis rats underwent colorectal instillation of trinitrobenzenesulphonic acid (TNBS) plus an equal amount of ethanol (test group) or physiological saline solution (control group). After 28, 35 and 42 days, standardized colorectal distensions were performed and the visceromotor reflex (VMR) of abdominal wall muscles was quantified by electromyographic recording. From day 28 onwards, EcN was administered in drinking water. RESULTS: After TNBS, a significant increase of VMR was observed compared with saline controls over all study days. Administration of EcN reduced the TNBS-induced hyperalgesia [EcN: 863+/-125 microV vs placebo: 1258+/-157 microV (P<0.05)] at day 35, while there were no significant alterations at any other study day. CONCLUSION: The EcN administration caused a significant reduction of VH. Whether EcN might play a role in the treatment of post-infectious functional bowel disorders remains to be investigated in further studies.

Specific proliferative and antibody responses of premature infants to intestinal colonization with nonpathogenic probiotic E. coli strain Nissle 1917.

The aim of this study was to analyze the influence of oral administration of E. coli Nissle 1917 on the systemic humoral and cellular immunity in premature infants. Thirty-four premature infants were colonized with E. coli Nissle 1917 in a randomized, placebo-controlled blinded clinical trial. Stool samples of infants were analyzed repeatedly for the presence of the administered strain. The proliferative response to bacterial antigens of E. coli origin was measured in whole blood of 34 colonized infants and 27 noncolonized controls. E. coli colonization induced a significant increase in the proliferation of blood cells cultivated with bacterial components of E. coli Nissle 1917 and another E. coli strain in colonized infants as compared with noncolonized controls. Significantly higher amounts of specific anti-E. coli Nissle 1917 antibodies (Ab) of immunoglobulin (Ig)A isotype and nonspecific polyclonal IgM were found in the blood of colonized infants compared to noncolonized placebo controls. We concluded that the oral application of E. coli Nissle 1917 after birth significantly stimulates specific humoral and cellular responses and simultaneously induces nonspecific natural immunity.

The impact of nonnormality on full information maximum-likelihood estimation for structural equation models with missing data.

A Monte Carlo simulation examined full information maximum-likelihood estimation (FIML) in structural equation models with nonnormal indicator variables. The impacts of 4 independent variables were examined (missing data algorithm, missing data rate, sample size, and distribution shape) on 4 outcome measures (parameter estimate bias, parameter estimate efficiency, standard error coverage, and model rejection rates). Across missing completely at random and missing at random patterns, FIML parameter estimates involved less bias and were generally more efficient than those of ad hoc missing data techniques. However, similar to complete-data maximum-likelihood estimation in structural equation modeling, standard errors were negatively biased and model rejection rates were inflated. Simulation results suggest that recently developed correctives for missing data (e.g., rescaled statistics and the bootstrap) can mitigate problems that stem from nonnormal data.

TP53 DNA contact mutations are selectively associated with allelic loss and have a strong clinical impact in head and neck cancer.

Recent studies have suggested that different mutation types within the core domain of the tumour suppressor protein p53, i.e. DNA contact mutations and structural mutations, confer different biological properties. We have analysed in 86 head and neck squamous cell carcinomas (HNSCC), whether these p53 mutation types have a differential clinical impact. Thirty-seven missense mutations were identified. Thirteen of these (36%) were DNA contact mutations, occurring in the L3 loop, in the H2 loop sheet helix motif, in the S10 beta strand and in Zinc binding residues. Microsatellite marker analysis revealed a selective association between these mutations and the loss of wild-type alleles (100% LOH vs 50% LOH in tumours with structural mutations; P=0.0034, Fisher's exact, 2-tailed). In comparison to structural mutations or to the absence of mutations in the core domain, DNA contact mutations were associated with higher tumour stages (84.6% vs 62%), a higher incidence of lymph node metastasis (91.7% vs 56%; P=0.014, Fisher's exact, 2-tailed), a shortened recurrence-free survival (8.1 months vs 23.7 months, P=0.047, log rank test) and overall survival (11 months vs 29.2 months; P=0.003, log rank test). The latter was also the case when only stage IV tumours were analysed (P=0.0055, log rank test). These data indicate that in HNSCC, TP53 DNA contact mutations confer a strong selection pressure to eliminate wild-type alleles, and that they result in an accelerated tumour progression and reduced therapeutic responsiveness.

Aberrant p21(CIP1/WAF1) protein accumulation in head-and-neck cancer.

p21(CIP1/WAF1) is an inhibitor of cyclin-dependent kinases and, in normal tissues including squamous epithelia, has been associated with cell-cycle exit and differentiation. As shown in this pilot study, however, the majority of head-and-neck squamous-cell carcinomas (HNSCC) display aberrant p21(CIP1/WAF1) expression: of 42 tumors analyzed by immunohistochemical staining, 28 (67%) over-expressed the p21(CIP1/WAF1) protein. Accumulation of p21(CIP1/WAF1) was independent of the histological grade of the tumors as well as the genetic status of the p53 gene. In many cases, most notably in poorly differentiated or undifferentiated HNSCC, p21(CIP1/WAF1)-positive cells were actively proliferating tumor cells, since they also expressed proliferating-cell nuclear antigen (PCNA) and Ki-67. Accumulation of p21(CIP1/WAF1) occurred through a post-transcriptional mechanism since, in contrast to immunohistochemical analysis of the p21(CIP1/WAF1) protein, in situ hybridization showed no increase of mRNA levels as compared with cells in normal mucosa (n = 25). Clinically, among the patients with p21(CIP1/WAF1)-over-expressing tumors, there was increased recurring disease (p = 0.03; chi2-test), shortened disease-free survival (p = 0.0019; log-rank test) and shortened overall survival (p = 0.0071; log-rank test). These in vivo data indicate that in many HNSCC, accumulated p21(CIP1/WAF1) is compatible with increased tumor-cell proliferation, and they provide preliminary evidence that p21(CIP1/WAF1) may be of prognostic and predictive significance.

Expression of mutated p53 occurs in tumor-distant epithelia of head and neck cancer patients: a possible molecular basis for the development of multiple tumors.

As in most other tumor types, expression of mutated or phenotypically altered p53 is a common occurrence in head and neck carcinogenesis. Since the prognosis for many head and neck tumor patients is severely affected by the occurrence of multiple primary and secondary tumors, we have analyzed the phenotype and genotype of p53 in squamous and respiratory epithelia either adjacent to or at significant distance from the primary tumors. Many tumor patients showed multifocal overexpression of the p53 protein in a variety of these epithelia. Overexpression of p53 correlated with increased proliferation and dedifferentiation, as demonstrated by immunohistochemistry and in situ hybridization using histone H3 and cytokeratin-specific probes. Polymerase chain reaction-single-strand conformation polymorphism analysis and sequencing of p53 DNA, amplified from these biopsies after immunostaining and microdissection, confirmed and extended these findings. We have identified different mutations in p53 in different tumor-distant epithelia from the same patients. The data indicate that mutation of p53 is an early event in head and neck carcinogenesis, preceding signs of overt neoplasia, and that different mutations in p53 in multiple foci may provide a molecular basis for the development of multiple tumors.

Renal nerves affect rate of achieving sodium balance in spontaneously hypertensive rats.

The spontaneously hypertensive rat (SHR) has an elevated efferent sympathetic nerve activity, suggesting that the renal handling of sodium and water may be altered. This study evaluated the renal neurogenic influence on the rate of achieving sodium balance in adult SHRs and Wistar-Kyoto (WKY) rats after either a step increase or step decrease in fixed sodium intake. Conscious, unrestrained rats with either innervated or denervated kidneys were initially placed on a low-sodium (0.3 mEq/d) or high-sodium (5.0 mEq/d) intake by intravenous infusion. Hourly urinary sodium excretion was determined 24 hours before and 72 hours after sodium intake had been increased from low to high or decreased from high to low. After either step change in fixed sodium intake, both innervated SHRs and innervated WKY rats achieved sodium balance within 24 hours. Similarly, the time course of achieving sodium balance was nearly identical between WKY rats with innervated and denervated kidneys after either switch in sodium intake. In SHRs receiving a step increase in sodium intake, both innervated and denervated kidneys increased urinary sodium excretion equally for 9 hours; however, at this time, innervated SHRs continued to increase sodium excretion rapidly, whereas denervated rats were delayed in a further response. Thus, innervated SHRs achieved sodium balance approximately 18 hours sooner than denervated SHRs. Differences in urinary sodium excretion did not result from concomitant changes in plasma renin activity or mean arterial pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro transformation of the tremorgenic mycotoxin verruculogen.

A quantity of 50 mg of crystalline verruculogen was prepared from batch cultures ofAspergillus fumigatus for the use in thein vitro studies with S-9 liver fractions, feces suspensions, and cultures ofEscherichia coli.Incubation of verruculogen with S-9 liver fractions from swine resulted in the transformation of the parent compound into TR-2 toxin and 3 other more polar.TR-2 toxin was also shown to be the main transformation product when incubat-ing verruculogen with fecal suspensions or pure cultures ofE. coli (0149: K88). Incubation times of more than 2 hours led to a complete degradation of verruculogen as well as TR -2 toxin.

Organization and nucleotide sequence of two chromosomal genes for rat cytochrome c oxidase subunit VIc: a structural and a processed gene.

A rat genomic library was screened with a cDNA probe coding for rat liver cytochrome c oxidase subunit VIc (COX-VIc). Out of 16 clones mapped, four inserts were different and did not overlap. Two inserts that gave strong hybridization signals were characterized by sequence analysis. These data show that we have cloned two different genes for rat COX-VIc. The rat COX-VIc-2 gene contains four exons and three introns. The 5' boundary of the first exon was mapped by primer extension experiments. The nucleotide sequence of the first three exons is identical to the sequence of the rat liver cDNA probe; thus, we can conclude that this gene is expressed in rat liver. The second gene (COX-VIc-1) is 88% homologous to the rat liver cDNA and contains an open reading frame of 228 nucleotides capable of coding for a full-length COX-VIc polypeptide of 76 amino acids. This predicted translation product is 79% homologous to the rat liver peptide. The absence of introns and other factors, such as the presence of a 13-bp direct repeat and a poly(A) addition signal, indicate that this locus is a processed gene which may have evolved from spliced mRNA intermediates.