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Biomedical subjects

G Richard

Publications and source records attributed to G Richard.

At least 55 records · Page 3Linked to original sources

The spectrum of pathogenic mutations in SPINK5 in 19 families with Netherton syndrome: implications for mutation detection and first case of prenatal diagnosis.

The Comèl-Netherton syndrome is an autosomal recessive multisystemic disorder characterized by localized or generalized congenital ichthyosis, hair shaft abnormalities, immune deficiency, and markedly elevated IgE levels. Life-threatening complications during infancy include temperature and electrolyte imbalance, recurrent infections, and failure to thrive. To study the clinical presentations of the Comèl-Netherton syndrome and its molecular cause, we ascertained 19 unrelated families of various ethnic backgrounds. Results of initial linkage studies mapped the Comèl-Netherton syndrome in 12 multiplex families to a 12 cM interval on 5q32, thus confirming genetic homogeneity of Comèl-Netherton syndrome across families of different origins. The Comèl-Netherton syndrome region harbors the SPINK5 gene, which encodes a multidomain serine protease inhibitor (LEKTI) predominantly expressed in epithelial and lymphoid tissues. Recently, recessive mutations in SPINK5 were identified in several Comèl-Netherton syndrome patients from consanguineous families. We used heteroduplex analysis followed by direct DNA sequencing to screen all 33 exons and flanking intronic sequences of SPINK5 in the affected individuals of our cohort. Mutation analysis revealed 17 distinct mutations, 15 of which were novel, segregating in 14 Comèl-Netherton syndrome families. The nucleotide changes included four non-sense mutations, eight small deletions or insertions leading to frameshift, and five splice site defects, all of which are expected to result in premature terminated or altered translation of SPINK5. Almost half of the mutations clustered between exons 2 and 8, including two recurrent mutations. Genotype-phenotype correlations suggested that homozygous nucleotide changes resulting in early truncation of LEKT1 are associated with a severe phenotype. For the first time, we used molecular data to perform prenatal testing, thus demonstrating the feasibility of molecular diagnosis in the Comèl-Netherton syndrome.

Adolescent↗

[Optical coherence tomography in geographic atrophy--a clinicopathologic correlation].

BACKGROUND: Optical coherence tomography was used for the examination of patients with geographic atrophy in different stages of age-related macular degeneration. Always compared with biomicroscopy and fluorescein angiography [4,7,8,10]. PATIENTS AND METHODS: 37 patients with geographic atrophy (n = 55 eyes) out of 150 with AMD (n = 169 eyes) were examined. The results of biomicroscopy, fluorescein angiography, optical coherence tomography and histological knowledge in age-related macular degeneration were studied. RESULTS: Fluorescein angiography always identified geographic atrophy and in 13.5% the findings additionally were similar to an occult choroidal neovascularisation with circular hyperpigmentation. Geographic atrophy shows a significant thinning of the neurosensory retina of 135 microns as an average in optical coherence tomography (p < 0.0005) which did not correlate to visual acuity. Typically an enhanced vertically sharp demarcated reflectivity of the choroid is found because of the lacking pigment epithelium. 43% of the geographic atrophies were identified by optical coherence tomography. 3 out of 55 eyes (5%) in optical coherence tomography only show macular holes additionally to geographic atrophy. CONCLUSION: A typical pattern of reflectivity is found by optical coherence tomography with enhanced reflectivity of the choroid because of lacking pigment epithelium and significant thinning of the fovea. Atypical macular holes moreover are found in 5% neither appearing in biomicroscopy nor in fluorescein angiography. An occult choroidal neovascularisation can be differentiated by optical coherence tomography from geographic atrophy because there is no spindle-like thickening of the pigment epithelium and no enhanced choroidal reflectivity. In borderline cases optical coherence tomography may be helpful and therapeutically decisive in differentiating geographic atrophy and occult choroidal neovascularisation and in detecting atypical macular holes.

Aged↗

trans-dominant inhibition of connexin-43 by mutant connexin-26: implications for dominant connexin disorders affecting epidermal differentiation.

Dominant mutations of GJB2-encoding connexin-26 (Cx26) have pleiotropic effects, causing either hearing impairment (HI) alone or in association with palmoplantar keratoderma (PPK/HI). We examined a British family with the latter phenotype and identified a new dominant GJB2 mutation predicted to eliminate the amino acid residue E42 (DeltaE42) in Cx26. To dissect the pathomechanisms that result in diverse phenotypes of dominant GJB2 mutations, we studied the effect of three Cx26 mutants (DeltaE42, D66H and R75W) identified in individuals with PPK/HI, and another (W44C) present in individuals with non-syndromic HI on gap junctional intercellular communication. We expressed mutant Cx26 alone and together with the epidermal connexins Cx26, Cx37 and Cx43 in paired Xenopus oocytes, and measured the intercellular coupling by dual voltage clamping. Homotypic expression of each connexin as well as co-expression of wild-type (wt) Cx26/wtCx43 and wtCx26/wtCx37 yielded variable, yet robust, levels of channel activity. However, all four Cx26 mutants were functionally impaired and failed to induce intercellular coupling. When co-expressed with wtCx26, all four mutants suppressed the wtCx26 channel activity consistent with a dominant inhibitory effect. However, only those Cx26 mutants associated with a skin phenotype also significantly (P<0.05) inhibited intercellular conductance of co-expressed wtCx43, indicating a direct interaction of mutant Cx26 units with wtCx43. These results demonstrate, for the first time, a trans-dominant negative effect of Cx26 mutants in vitro. Furthermore, they support a novel concept suggesting that the principal mechanism for manifestation of dominant GJB2 mutations in the skin is their dominant interference with the function of wtCx43. This assumption is further corroborated by our finding that Cx26 and Cx43 focally colocalize at gap junctional plaques in affected skin tissue of two carriers of DeltaE42.

Adolescent↗

The fate of heterotopically grafted neural precursor cells in the normal and dystrophic adult mouse retina.

PURPOSE: To study the integration and differentiation of heterotopically transplanted neural precursor cells in the retina of adult mouse mutants displaying apoptotic degeneration of photoreceptor cells. METHODS: Neural precursor cells were isolated from the spinal cord of transgenic mouse embryos ubiquitously expressing enhanced green fluorescent protein. Cells were expanded in vitro and transplanted into the retina of adult wild-type and age-matched beta2/beta1 knock-in mice. Beta2/beta1 knock-in mutants display apoptotic death of photoreceptor cells and were generated by placing the cDNA of the beta1 subunit into the gene of the beta2 subunit of Na,K-ATPase. The integration and differentiation of grafted cells in recipient retinas was studied 1 or 6 months after transplantation. RESULTS: Mutant retinas contained more donor-derived cells than wild-type hosts. Moreover, in mutants, donor cells integrated into deeper retinal layers. In both genotypes, grafted cells differentiated into astrocytes and oligodendrocytes. Only a few ganglion cell axons were myelinated by donor-derived oligodendrocytes 1 month after transplantation, whereas extensive myelination of the nerve fiber layer was observed 6 months after transplantation. Unequivocal evidence for differentiation of grafted cells into neurons was not obtained. CONCLUSIONS: Heterotopically transplanted neural precursor cells are capable of integrating, surviving, and differentiating into neural cell types in normal and dystrophic retinas of adult mice. The particular environment of a pathologically altered retina facilitates integration of transplanted precursor cells. In principle, neural precursors may thus be useful to substitute for or replace dysfunctional or degenerated cell types. Results of the present study also indicate that replacement of retinal cell types is likely to require more appropriate donor cells, such as retinal precursor cells.

Actins↗

[Programmed cell death (apoptosis) in excised subretinal neovascularization].

BACKGROUND: The occurrence of apoptosis in choroidal neovascularizations (CNV) has only been described previously in a few cases. However, little is known about the extent and function of apoptosis. We analyzed the incidence of apoptosis in order to find new options in the therapy of CNV. Particular attention was given to the length of time that the process existed before surgery in patients with age-related macular degeneration (AMD). PATIENTS AND METHODS: In 34 patients (18 women, 16 men) ranging in age from 20 to 91 years (mean 70.3 years), CNV was detected by angiography. The majority of patients (n = 29) with CNV had AMD, which in most patients affected both eyes. In two patients CNV was due to post-traumatic proliferative vitreoretinopathy, in two other patients CNV occurred after a penetrating bulbus injury, and in one patient pseudoxanthoma elasticum was found. Thirty-four CNV membranes were excised by pars plana vitrectomy via retinotomy, fixed in formalin and embedded in paraffin. Sections were stained with PAS and H&E and examined microscopically. The in situ cell-detection kit (TUNEL) was used for immunohistochemical detection of apoptotic cells from fragmented DNA. RESULTS: In 74% (n = 25) of all patients and in 76% (n = 22) of patients with AMD, disseminated or focal apoptotic cells were detectable in the connective tissue, retinal pigment epithelium and in the vascular endothelium. In AMD patients with apoptotic cells, the SNV existed 1-12 months (mean 5.5 months) before surgical intervention. In patients without apoptotic cells, the CNV were present 5-36 months (mean 17 months) before surgery. At the time of surgery, the loss of reading ability had on average been going on 3 months; in patients with apoptotic cells, the mean time from the onset of this symptom was 2.4 months (1 week to 11 months); and in patients without apoptotic cells it was 5 months (1-12 months). CONCLUSION: Programmed cell death as a regulating pathomechanism was observed in all tissue parts of CNV with variable extension. In patients with AMD, a correlation existed between the length of time that CNV existed before surgery and the incidence of apoptotic cells. Apoptosis was found more frequently in recent CNV than in long-standing lesions. The various activity found in CNV suggests that the success of treatment may depend on the moment of intervention. To characterize the role of apoptosis further inducing and inhibiting factors have to be analyzed.

Adult↗

[Endophthalmitis after cataract surgery: predisposing factors, infectious agents and therapy].

BACKGROUND: Infectious exogenous endophthalmitis is a serious complication after cataract surgery. Despite modern pharmacological and surgical methods, its treatment is still difficult. PATIENTS AND METHODS: In a retrospective study the records of all patients treated for endophthalmitis following cataract extraction at the Department of Ophthalmology of the University Hospital in Hamburg between January 1989 and December 1997 were assessed. RESULTS: Of 36 patients treated for endophthalmitis, 29 (80.6%) had been referred. In 14 (38.9%) of these 29 patients endophthalmitis had occurred after outpatient cataract surgery. Seven patients (19.4%) had been treated as inpatients at the University Eye Hospital Eppendorf. Vitrectomy was performed in 80.6% of the cases. An infectious agent was isolated from 50% of diagnostic probes. The most common organism isolated were coagulase-negative staphylococci (4 cases). Predisposing factors for the development of endophthalmitis were diabetes (27.8%), intraoperative loss of vitreous (19.4%), application of systemic steroids (13.9%) and wound dehiscence (11.1%). Of 27 patients (75%) followed up, 16 (59.3%) had a final visual acuity of 20/400 or better (mean 20/40). An enucleation had to be performed in 4 patients (13.8%). CONCLUSION: In this study, approximately 60% of patients with endophthalmitis following cataract extraction had their globes preserved and a good visual outcome after appropriate surgical interventions.

Adrenal Cortex Hormones↗

[Ultrasound biomicroscopy in pigmentary glaucoma].

PURPOSE: To evaluate the anatomical relationships of the iris in pigmentary glaucoma before and after laser iridotomy and to evaluate the effect on intraocular pressure. METHODS: Ultrasound biomicroscopy (UBM, Humphrey-Zeiss) of the anterior segment was performed in 28 eyes of 28 patients (20 male, 8 female, mean age 43 years, mean untreated IOP 24.3 mmHg) with pigmentary glaucoma before and after laser iridotomy. The slope of intraocular pressure was documented. Mean follow-up was 9 months. For statistical analysis the Wilcoxon test was used. RESULTS: Ten out of 28 eyes showed iridozonular contact and concavity of the midperipheral iris. Laser iridotomy resulted in a significant pressure drop (P < 0.05) in these 10 eyes (24.6 mmHg to 18.3 mmHg). Eighteen eyes, however, did not show iridozonular contact and intraocular pressure did not drop sufficiently (P > 0.05; 25.1 mmHg to 23.1 mmHg) after laser iridotomy. CONCLUSION: The results show that iridozonular contact does not exist in every patient with pigmentary glaucoma. Therefore, it seems possible that more than one pathogenic mechanism is involved in pigmentary glaucoma. In patients with iridozonular contact, however, laser iridotomy significantly reduces intraocular pressure.

Adult↗

[Prolonged wound healing after perforating keratoplasty. CPAP hyperbaric ventilators for sleep apnea as a risk factor].

Nasal hyperbaric respiration devices used at night to counteract sleep apnea may represent a concealed cause of delayed superficial wound healing following perforating keratoplasty. We report the cases of two patients whose respirators produced a continuous flow of pressurized air (nCPAP) onto the operated eyes, leading to conjunctival hyperemia, recurrent corneal erosion, infiltration of the puncture track and the cornea, and superficial punctate keratopathy. Microbiological investigation revealed no clearly identifiable pathogens. Healing improved consistently only after the pressurized air system had been modified so that no air escaped from the nostrils and the patients had been instructed in disinfection of the moisturizing system.

Adult↗

Effect of perfluorodecalin on human retinal pigment epithelium and human corneal endothelium in vitro.

BACKGROUND: Perfluorocarbon liquids are useful intraoperative tools in complicated vitreoretinal surgery. They are usually removed at the end of the procedure, but small amounts may remain in the eye. Recently, contradictory results have been reported on the damage in association with residual perfluorocarbon liquids in the eye. This study examined the effects of perfluorodecalin on human retinal pigment epithelium and corneal endothelium in vitro. METHODS: Vitality and proliferative capacity of cell cultures were measured after incubation with perfluorodecalin. Vitality of cell cultures were measured using the Life-Dead assay. Cell proliferation was determined by measuring incorporation of 5-bromo-2'-deoxyuridine into cellular DNA. Furthermore, endothelium of organ-cultured human corneas was examined after incubation with perfluorodecalin by photodocumentation. RESULTS: Both cell types showed less extinctions in the Life-Dead assay after incubation with perfluorodecalin. After removing perfluorodecalin from the cultures, cells showed the same capacity of proliferation as the control cells. Compared to control corneas, perfluorodecalin induced a decrease in endothelial cell density. In four corneas, endothelial cell necrosis was observed. CONCLUSION: Decreasing extinctions in the Life-Dead assay after incubation with perfluorodecalin can be interpreted as showing a decreasing amount of vital cells. Because cell proliferation showed no significant changes the results suggest that perfluorodecalin may not be directly toxic to cells in vitro. It may exert an indirect or mechanical effect on cell function by impeding the normal metabolic exchange between endothelium and medium. Based on these results perfluorodecalin should be completely removed after operation.

Cell Count↗

The spectrum of mutations in erythrokeratodermias--novel and de novo mutations in GJB3.

Intercellular channels in skin are a complex and functionally diverse system formed by at least eight connexins (Cx). Our recent molecular studies implicating Cx defects in inherited skin disorders emphasize the critical role of this signaling pathway in epidermal differentiation. Erythrokeratodermia variabilis (EKV) is an autosomal dominant genodermatosis with a striking phenotype characterized by the independent occurrence of transient localized erythema and hyperkeratosis. The disease maps to 1p34-p35, and recently we identified the causative gene GJB3 encoding Cx31. We have now investigated GJB3 in two families and three sporadic cases with EKV, and report three new heterozygous mutations. In a sporadic case, we detected a mutation leading to substitution of a conserved phenylalanine (F137L) in the third transmembrane domain, which likely interferes with the proper assembly or gating properties of connexons. In another family, all three affected individuals carried two distinct mutations on the same GJB3 allele. However, only a de novo heterozygous missense mutation replacing arginine 42 with proline (R42P) co-segregated with the disease, while a 12 bp deletion predicted to eliminate four amino acid residues in the variable carboxy terminal domain of Cx31 was also found in clinically unaffected relatives but not in 90 unaffected controls. Including the previously published mutations, in toto, five different missense mutations have now been detected in 6 out of 17 families investigated by our laboratory, all of which presumably affect the cytoplasmic amino terminal and transmembrane domains of Cx31. In contrast, two mutations linked to progressive high-tone hearing impairment were located in the second extracellular domain, suggesting that the character and position of Cx mutations determine their phenotypic expression in different tissues. However, the phenotypic spectrum of GJB3 mutations seems not to include progressive symmetric erythrokeratodermia, another dominant genodermatosis with overlapping features, since no mutations were found in six unrelated families tested.

Connexins↗

Optical coherence tomography in uveitis patients.

PURPOSE: To evaluate optical coherence tomography in allergy-prone uveitis patients. METHODS: Thirty-four patients (43 eyes) with posterior uveitis (31 eyes) and intermediate uveitis (12 eyes) were evaluated by fluorescein angiography, indocyanine green angiography, and optical coherence tomography. Follow-up examinations used optical coherence tomography in allergy-prone patients. RESULTS: Optical coherence tomography identified epiretinal membranes, which were removed surgically (three eyes); persistent cystoid macular edema, which resolved with cytotoxic treatment (12 eyes); and juxtafoveolar membranes, which were treated by diode laser (six eyes) and excision (two eyes). CONCLUSION: Optical coherence tomography may provide useful information on complications developing in uveitis patients.

Adult↗

Detection of HPV-20, HPV-23, and HPV-DL332 in a solitary eyelid syringoma.

PURPOSE: To report evidence of many human papillomavirus types occurring in a solitary syringoma clinically appearing as a papilloma. METHODS: A 57-year-old man presented with a 10-year history of an upper eyelid tumor. Histopathology, human papillomavirus-nested polymerase chain reaction, human papillomavirus-DNA cloning into vector pCR2.1, sequencing, and computer-assisted evaluation were performed. RESULTS: Histopathology demonstrated a solitary benign syringoma. HPV-20 and HPV-23 were present in one clone each, and HPV-5-related HPV-DL332 was present in 9 clones. CONCLUSION: Many human papillomavirus types may be detected in an ocular syringoma.

DNA Primers↗

Ultra-rapid categorisation of natural scenes does not rely on colour cues: a study in monkeys and humans.

In a rapid categorisation task, monkeys and humans had to detect a target (animal or food) in briefly flashed (32 ms) and previously unseen natural images. Removing colour cues had very little effect on average performance. Impairments were restricted to a mild accuracy drop (in some human subjects) and a small reaction time mean increase (10-15 ms) observed both in monkeys and humans but only in the detection of food targets. In both tasks, accuracy and latency of the fastest behavioural responses were unaffected, suggesting that such ultra-rapid categorizations could depend on feed-forward processing of early coarse achromatic magnocellular information.

Adult↗

Connexins: a connection with the skin.

The intercellular signaling system mediated by connexin channels is crucial for maintaining tissue homeostasis, growth control, development, and synchronized response of cells to stimuli. This review summarizes the structure, assembly, and properties of the components of the complex and diverse connexin system, and their biological functions in skin. The importance of gap junctional intercellular communication for normal development and differentiation of human epidermis as well as the hearing function of the inner ear is illustrated by the examples of erythrokeratodermia variabilis and palmoplantar keratoderma associated with hearing loss. These autosomal dominant inherited disorders are caused by germline mutations in the connexin genes GJB3 (encoding connexin-31) and GJB2 (encoding connexin-26), respectively. Recent functional studies of individual connexin mutations suggest that they may exert a dominant inhibitory effect on normal connexin channel function and perturb gap junctional intercellular communication, resulting in phenotypic manifestation in patients with these disorders.

Cell Communication↗

Connexin mutations associated with palmoplantar keratoderma and profound deafness in a single family.

Recently, mutations in two gap junction genes, GJB2 and GJB3 (encoding Connexin 26 and Connexin 31, respectively), have been shown to underlie either inherited hearing loss and skin disease or both disorders. In this study, we have extended our analysis of a small family in which palmoplantar keratoderma and various forms of deafness is segregating. In addition to the previously described sequence variant M34T in GJB2, two other sequence variants were identified: D66H also in GJB2 and R32W in GJB3. As D66H segregated with the skin disease, it is likely to underlie the palmoplantar keratoderma. The other two gap junction variants identified may contribute to the type of hearing impairment and the variable severity of the skin disease in the family.

Connexin 26↗