Molecular machinery providing copper bioavailability for spermatozoa along the epididymial tubule in mouse.

Progressive functional maturation of spermatozoa is completed during the transit of these cells through the epididymis, a tubule structure connecting a testicle to a vas deferens. Epididymal epithelial cells by means of their secretory and absorptive functions determine a highly specialized luminal microenvironment containing multiple organic and inorganic components. The latter include copper ions, which due to their redox properties are indispensable for critical homeostatic processes occurring in spermatozoa floating in different part of epididymis but can be potentially toxic. Main purpose of our study was to determine epididymal region-dependent expression and localization of copper transporters ensuring a tight control of copper concentration in epididymal fluid. We also aimed at identifying proteins responsible for copper uptake by spermatozoa and verifying whether this process is coordinated with copper supply to superoxide dismutase 1 (SOD1), a copper-dependent antioxidant enzyme. Our study identifies two ATPases-ATP7A, ATP7B and Slc31a1, major copper importers/exporters depending on their differential expression on epididymal polarized epithelial cells of the caput, corpus, and cauda. Next, ceruloplasmin seems to be a chief protein transporting copper in the epididymal fluid and providing this biometal to spermatozoa. The entry of copper to germ cells is mediated by Slc31a1 and is correlated with both expressions of copper chaperone for superoxide dismutase (CCS), copper chaperone directly providing copper ions to SOD1 and with the expression and activity of the latter. Our results outline a network of cooperating copper binding proteins expressed in epididymal epithelium and in spermatozoa that orchestrate bioavailability of this microelement for gametes and protect them against copper toxicity.

[Analysis of 100 cases of retinal detachment treated with conventional surgery in the years 1997-1998]. / Analiza 100 przypadków odwarstwienia siatkówki leczonych konwencjonalnymi metodami w latach 1997-1998.

PURPOSE: The observed increasing number of severe cases of retinal detachment was the ground of the attempt to state the causes of this phenomenon. MATERIAL AND METHODS: Retrospective evaluation of 100 consecutive cases of retinal detachment operated on with conventional methods between September 1997 and October 1998 was performed. The following data were analysed: sex and age of patients, period between the first symptoms and time of diagnosis and surgery. The clinical characteristics of retinal surgery comprise: extent of detachment, macular involving, type and number of breaks, degenerations and PVR. RESULTS: In the majority of cases very severe retinal detachments were observed, including 3-4 quadrants, with large tears or multiple breaks, with PVR B or C. Duration of the detachment ranged from several days in very few cases to several months, on average lasting several weeks. Attachment of the retina was achieved in 80 cases, improvement of the visual acuity in 47. CONCLUSION: Our data indicate the necessity to improve the early diagnosis of retinal detachment and changes predisposing to its development.

[Arteriovenous communication of the retina--case report]. / Polaczenie tetniczo-zylne siatkówki--opis przypadku.

In this paper I have described a rare case of typical arteriovenous communication of the retina (ACVR), also called racemose haemangioma, but with some atypical traits. A case of a young white 22-year-old woman is presented. AVCR was recognised in her left eye, II stage according to Archer's classification, therefore neither intracranial haemangioma nor vascular decompensation of retina was observed. In the standard ophthalmic examination, including automated perymetry and fluorescein angiography, the following phenomena were not seen yet or they were rarely described: pigmented muffs on some changed vessels, mainly nasal location of AVCR, parallel AVCR existed between both great and precapillar vessels and macular changes: yellowish ring surrounded the central fovea. Visual acuity of the affected eye was 1.0 and the patient did not complain about it.

Analysis of the trinucleotide CAG repeat from the human mitochondrial DNA polymerase gene in healthy and diseased individuals.

The human nuclear gene (POLG) for the catalytic subunit of mitochondrial DNA polymerase (DNA polymerase gamma) contains a trinucleotide CAG microsatellite repeat within the coding sequence. We have investigated the frequency of different repeat-length alleles in populations of diseased and healthy individuals. The predominant allele of 10 CAG repeats was found at a very similar frequency (approximately 88%) in both Finnish and ethnically mixed population samples, with homozygosity close to the equilibrium prediction. Other alleles of between 5 and 13 repeat units were detected, but no larger, expanded alleles were found. A series of 51 British myotonic dystrophy patients showed no significant variation from controls, indicating an absence of generalised CAG repeat instability. Patients with a variety of molecular lesions in mtDNA, including sporadic, clonal deletions, maternally inherited point mutations, autosomally transmitted mtDNA depletion and autosomal dominant multiple deletions showed no differences in POLG trinucleotide repeat-length distribution from controls. These findings rule out POLG repeat expansion as a common pathogenic mechanism in disorders characterised by mitochondrial genome instability.

[Risk factors for age-related macular degeneration]. / Czynniki ryzyka zwyrodnienia plamki zwiazanego z wiekiem.

PURPOSE: To present the results of examinations of the risk factors for age-related macular degeneration (AMD) carried out in the last 3 years. MATERIAL AND METHODS: Our clinical material comprised 119 patients, 69 women and 50 men, aged 43-85, mean age 70 years. Using classification according to the worse eye, there were 20 patients with drusen, 27 with atrophic changes and 72 with exudative form and AMD. The following risk factors were evaluated: age, sex, body mass index, history of general medical conditions, cigarette smoking, sun exposure, family history of AMD and ocular conditions such as iris color, lens opacities, hyperopia, gerontoxon and changes in retinal vessels. RESULTS: The significant relationships were found between the development of AMD and the age of patients, as well as between the advanced forms of AMD and the history of cardiovascular diseases and sclerotic changes in retinal vessels, 87% of examined patients have light iris and 52% body mass index above 26. CONCLUSIONS: The studies confirmed the role of age in the development of AMD and indicated cardiovascular disturbances, increased body mass index and light iris as the possible risk factors for AMD that are most worth further studying. The special attention should be also paid to drusen as the risk factor of AMD.

[Conventional methods in retinal detachment surgery]. / Konwencjonalne metody w chirurgii odwarstwienia siatkówki.

PURPOSE: To evaluate the usefulness of conventional methods in retinal detachment (RD) surgery basing on the relationship between factors characterizing clinical picture of the RD, the type and course of surgical procedures and the retinal reattachment. MATERIALS AND METHODS: 252 patients (252 eyes), operated on in the last 5 years in our clinic were enrolled in the studies. There were 120 men and 132 women, aged 9 to 83, mean 53. In all cases scleral buckling procedures with silicone band or sponge and cryocoagulation of the breaks were used. The relationship between retinal reattachment and the following parameters were examined: visual acuity, retinal degenerations, myopia, extent of the detachment, number and kinds of breaks, macular involvement, state of vitreous, PVR, intraocular pressure, extent of scleral buckling, subretinal fluid drainage and intra- or post-operative complications. RESULTS: Retinal reattachment was achieved in 82% of the eyes with one operation and additionally in 7% after reoperation. The significant relationship was found between: visual acuity, intraocular pressure, extent of the detachment, number of breaks, PVR, extent of scleral buckling and retinal reattachment. CONCLUSION: PVR is a significant cause of failure in RD surgery with conventional methods and PVR grade C is the threshold beyond which the percentage of reattachments decreases to about 50%. Value of other risk factors, determining severity of RD is not certain for prognosis. Cerclage with silicone band is a method of choice in the majority of RD with PVR grade B and C. Our observations do not confirm the reports of severe complications caused by cerclage, which might be connected with excessive tightening of the band.

Yeast mitochondrial NAD(+)-dependent isocitrate dehydrogenase is an RNA-binding protein.

We have previously described the characterisation of an abundant mitochondrial protein (p40) that binds specifically to 5'-untranslated leaders of mitochondrial mRNAs in yeast. p40 consists of two polypeptides with M(r) of 40 and 39 kDa. Limited sequence analysis of p40 identifies it as the Krebs cycle enzyme NAD(+)-dependent isocitrate dehydrogenase (Idh). Both enzyme and RNA-binding activities are specifically lost in cells containing disruptions in either IDH1 or IDH2, the nuclear genes encoding the two subunits of the enzyme, thus conclusively identifying p40 as Idh and showing that both activities are dependent on the simultaneous presence of both subunits. Although we still must ascertain whether and how either function of Idh is regulated and whether the two functions are compatible or mutually exclusive, this combination of dehydrogenase activity and RNA-binding in a single protein may be part of a general regulatory circuit linking the need for mitochondrial function to mitochondrial biogenesis.

[Value of silicone sponge in retinal detachment surgery]. / Wartosc gabki sylikonowej w operacjach odwarstwienia siatkówki.

Lincoff's development of silicone sponge and substitution cryosurgery for diathermy in the mid-1960s, was the beginning of the minimized surgery for retinal detachment, limited to the surgery of the breaks. The aim of this paper is to present our experiences with these kinds of operations, with special attention to the complications causing the necessity of removing the sponge. Our studies concerned the patients who underwent retinal detachment surgery, in the Ophthalmological Clinic in Cracow in the years 1983-1992, using scleral buckling procedures with silicone sponge. In 152 cases circumferential and in 36 radial local scleral buckling procedures were performed; in 30 cases local buckling was added to encirclement with silicone band. The complications, attributed to the sponge, were observed in 9.7% of cases. Anterior segment inflammatory reactions, often sterile were found in 15 eyes, in 7 of them it was necessary to remove the sponge. In 13 eyes the sponge was removed on account of its extrusion. In none of the cases removal of the sponge influenced the retinal attachment. Our studies confirmed, that silicone sponge is a very good material for scleral buckling procedures.

[Drainage of subretinal fluid in our clinical studies]. / Drenaz plynu podsiatkówkowego w swietle wlasnych badan.

The studies comprised 531 patients (544 eyes) who underwent retinal detachment surgery in the Ophthalmological Clinic in Cracow, in the years 1983-1992. Drainage of subretinal fluid was performed in 54.6% of the eyes and in this group attachment of the retina was achieved in 72.4% of cases; in the group of patients with nondrainage operations it was 74.5%. Intraocular haemorrhages, the only complications, which may be attributed to the drainage of subretinal fluid were found in 18.9% of eyes with drainage and in 8.1% without; this difference was statistically significant (p < 0.01). Statistically significant relationship (p < 0.01) was also found between subretinal fluid drainage and severity of the retinal detachment. The results showed that there were no significant relationships between drainage of subretinal fluid and the results of surgery and that it is indicated first of all in severe cases of retinal detachment in which method of choice is an encirclement procedure.

[Relationship between results of retinal detachment surgery and method of attachment]. / Zaleznosc miedzy wynikami leczenia operacyjnego odwarstwienia siatkówki a rodzajem blokady otworów.

Basing on the clinical material of the years 1984-1991 the authors have compared the results of retinal detachment surgery in two groups of patients, depending on the method of adhesion. Diathermy was used in 182 eyes (group I) and cryosurgery in 274 (group II). Retinal attachment was achieved in 69% of the eyes in group I and in 74% in group II. Inadequate closing of the breaks was the cause of failure in 22.8% of the cases in group I and in 30.6% in group II. Various changes of the macula were observed in 31% of the eyes in group I and in 24.5% in group II. Statistically significant relationships were found between method of adhesion and: development of PVR (p < 0.05), appearance of pigmentary changes in the eye ground (p < 0.01) and cicatrization of the breaks (p < 0.01). Stronger cicatrization was observed after cryosurgery as well as the pigmentary changes in the fundus which have not influenced the results. The studies revealed that no significant differences depending on the method of adhesion were found in results of retinal detachment surgery.

[A case of Stickler's syndrome]. / Przypadek zespolu Stickler'a.

The authors presented a case of Stickler's syndrome diagnosed in 56-year-old woman referred to the clinic because of cataract in both eyes. After surgery typical vitreo-retinal changes mainly extensive lattice degenerations, pigmentary lesions and preretinal membranes in vitreous were found. Orofacial and musculoskeletal abnormalities confirmed diagnosis.

[Use of photocoagulation during surgery for retinal detachment]. / Zastosowanie fotokoagulacji w operacjach odwarstwienia siatkówki.

On the basis of a 10-years clinical material comprising 95 eyes operated for retinal detachment in the Department of Ophthalmology in Krakow the authors evaluated the usefulness of xenon and laser coagulation as a method complementing the blockade of the retinal holes. It had been used in 18 p.c. of all operated eyes attaining in this group of cases the re-apposition of the retina in 61 p.c. The efficacy of the coagulation depended on the adherence of the retina and showed itself as formation during the surgery of visible foci and the development of a distinct, pigmented scar. There were no complications connected with photocoagulation as well the xenon as the laser one; all this encourages to employ both these methods.

[Probability of successful operation in very severe retinal detachment using scleral buckling procedure]. / Prawdopodobienstwo powodzenia operacji w bardzo ciezkich odwarstwieniach siatkówki przy stosowaniu technik wglabiajqcych twardówke.

The aim of this study was to study the efficacy of operation of scleral invagination in the treatment of very severe retinal detachment with proliferative vitreoretinopathy (PVR). The investigations comprised 36 eyes operated in the Department of Ophthalmology in Krakow in the period 1986-1990. The authors performed in 24 patients a encircling operation with a silicone band, in 12-meridional or parallel extrascleral silicone sponge implants. Reattachment of the retina was achieved in 58 p.c. of eyes and an improvement of the visual acuity in 47 p.c. The performed analysis showed a substantial dependence of the attained effect of surgery on the grade of advancement of the PVR; it enabled to ascertain that the limit of possibilities of the scleral buckling procedure determines PVR stadium C3.

[Late results of surgical treatment of retinal detachment by the cerclage method using a silicone band]. / Odlegle wyniki leczenia operacyjnego odwarstwienia siatkówki metoda opierscienienia tasma sylikonowa.

Presented are the results of control examination performed in 121 patients in the period of 3 to 8 years after retinal detachment operation by the silicone band encircling method. Evaluated were: the anatomical reattachment, function of the eye and the cosmetic effect. The retina reattached in 63.2 p.c. of eyes. The visual acuity for far was at least 0.1 in over one half (55 p.c.) of eyes and for near 2.0 or more in 78.5 p.c. Among the persons examined by means of Amsler's test and Panel D-15 test a correct response was attained by -- respectively -- 44.7 p.c. and 76.1 p.c. of the examinees. A very good cosmetic effect was achieved in 44.9 p.c. and a good one in 47.2 p.c. of eyes.

Determinants of correct res site alignment in site-specific recombination by Tn3 resolvase.

In site-specific recombination reactions catalyzed by Tn3 resolvase, the right and left arms of the res site are always religated to the correct partner. This poses the problem of how resolvase aligns the two sites correctly for the cleavage/religation reaction. We show that the "accessory" binding subsites II and III of res are important for correct alignment of the adjoining crossover subsite (subsite I). Deletion of subsites II and III from one of the two res sites removes a barrier to recombination between incorrectly aligned crossover subsites. Correct alignment does not require any DNA sequence asymmetry in the crossover subsite, DNA supercoiling, or covalent linkage of the two res sites. Our results suggest that correct subsite I alignment is determined by local, resolvase-mediated interactions of subsites II and III of both partners, consistent with a current model of the synapse. Surprisingly, the topological selectivity for intramolecular resolution in a supercoiled substrate does not require subsites II and III in both recombination partners.

L-malate oxidation by the electron transport fraction of Azotobacter vinelandii.

The membrane-bound l-malate oxidoreductase of Azotobacter vinelandii strain O was found to be a flavoprotein-dependent enzyme associated with the electron transport system (R(3)) of this organism. The particulate R(3) fraction, which possessed the l-malate oxidoreductase, carried out the cyanide-sensitive oxidation of l-malate, d-lactate, reduced nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate, succinate, cytochrome c, tetramethyl-p-phenylenediamine, and p-phenylenediamine, with molecular O(2) as the terminal electron acceptor. d-Malate was not oxidized, but l-malate was oxidized to oxalacetate. Phenazine methosulfate (PMS), vitamin K(3), K(3)Fe(CN)(6), nitro blue tetrazolium, and dichloroindophenol all served as good terminal electron acceptors for the l-malate oxidoreductase. Cytochrome c was a poor electron acceptor. Extensive studies on the l-malate oxidase and PMS and K(3) reductases revealed that all were stimulated specifically by flavine adenine dinucleotide and nonspecifically by di- or trivalent cations, i.e., Ca(++), Ba(++), Mn(++), Mg(++), Fe(+++), Ni(++), and Al(+++). All these activities were markedly sensitive to ethylenediaminetetraacetate (EDTA). The V(max) values for the l-malate oxidase, PMS, and vitamin K(3) reductases were, respectively, 3.4, 15.1, and 45.5 mumoles of substrate oxidized per min per mg of protein at 37 C. Spectral studies revealed that the Azotobacter R(3) flavoprotein and cytochromes (a(2), a(1), b(1), c(4), and c(5)) were reduced by l-malate. l-Malate oxidase activity was sensitive to various inhibitors of the electron transport system, namely, p-chloromercuriphenylsulfonic acid, chlorpromazine, 2-n-heptyl-4-hydroxyquinoline-N-oxide, antimycin A, and KCN. Minor inhibitory effects were noted with the inhibitors 4,4,4-trifluoro-1-(2-thienyl)-1,3-butanedione, rotenone, and Amytal.