Effect of hen's egg yolk on capacitation and acrosome reaction of diluted canine spermatozoa.

UNLABELLED: The aim of this study was to investigate the influence of progesterone, cholesterol and calcium (Ca(2+)) in an egg-yolk-containing extender on capacitation and acrosome reactions (AR) of diluted canine spermatozoa during 4 days of cooled-storage. For this purpose, we first investigated the effect of supplementation of a Tris-citrate-fructose buffer (TCF) with progesterone in a final concentration of 0.1, 0.2 and 1.0 microg progesterone/ml TCF-diluted semen. We then compared the effects of TCF and the same buffer-containing 20% egg yolk (TCF-EY). In egg yolks and the TCF-EY, progesterone was measured by enzyme immunoassay, cholesterol by enzymatic colorimetry and Ca(2+) by flame atomic absorption spectrophotometry. For both experiments, ejaculates from eight dogs were used. For the comparison of diluents, one ejaculate was divided and one half diluted with TCF, the other with TCF-EY. One half of each TCF- and TCF-EY-diluted sample was evaluated immediately (D1), the other after storage for 4 days at +4 degrees C (D4). In diluted semen, motility and viability were measured by a computer assisted sperm analyzer (CASA; Sperm Vision, Minitüb, Germany), capacitation and AR were evaluated with a modified chlortetracycline assay (CTC) and the AR additionally by flow cytometry. RESULTS: Supplementation of progesterone revealed, that between D1 and D4, total and progressive motility decreased with all progesterone concentrations, while viability as well as percentage of capacitated and acrosome reacted spermatozoa stayed constant. Progesterone-, cholesterol- and Ca(2+) concentrations in egg yolks were 524.8+/-131.4 ng/g, 13.9+/-2.03 mg/g and 1.27+/-0.17 mg/g, respectively. In the TCF-EY-diluent, the respective values were 210.9 ng/g, 2.52 mg/g and 1.1mg/g. In TCF-semen, at D1, motility and viability were significantly higher than in TCF-EY-samples (p<0.05), however at D4, no significant differences were detectable. Further, in TCF-semen, percentages of spermatozoa with intact membranes decreased significantly (p<0.05) and capacitated spermatozoa increased (p<0.05), which was not seen in TCF-EY-samples. In all samples, low percentages of AR were detected and after 4 days, the highest value of AR in TCF-EY-samples was 5.3% on average, as detected by flow cytometry. We therefore conclude that progesterone from egg yolk in routine extenders does not substantially influence semen longevity or AR of canine semen during cold-storage for 4 days. In contrary, egg yolk seems to prevent a significant increase in capacitated spermatozoa.

Antegrade balloon dilatation of nasolacrimal duct obstruction in adults.

AIMS: To determine the efficacy of antegrade balloon dilatation of postsaccal lacrimal stenosis in adults. METHODS: Balloon dilatation was performed in a series of 30 patients with complete nasolacrimal duct obstructions and epiphora. Obstruction was diagnosed by canalicular irrigation and transcanalicular endoscopic examination of the lacrimal pathway. Except for four cases in which general anaesthesia was applied, the procedure was performed under local anaesthesia. The Lacricath balloon catheter set was used. Silicone intubation was performed simultaneously. The time at which the tubes were removed depended on the findings at postoperative follow up but was, at the earliest, 3 months postoperatively. RESULTS: Success was objectified by irrigation and was evaluated subjectively at each follow up examination according to Munk's scale. In all cases the procedure could be performed with subsequent silicone intubation. Three months postoperatively 89.9% of all cases were positive on simple irrigation, and subjective success was also registered (Munk's grade 0 or 1). At 6 months 70% of all cases were positive on irrigation, again with subjective success (Munk's grade 0 or 1). One year postoperatively 73.3% of all procedures showed subjective success (two successful redilatations would raise the success rate to 79.9%). CONCLUSION: Retrograde as well as antegrade dilatation has been reported to be more or less successful in partial nasolacrimal obstruction. Although the procedure is used as primary treatment in cases of complete obstruction, it can still be performed under local anaesthesia on an outpatient basis. Long term observation will be required to prove the sustained effect of this procedure.

Bacteriology and antibiotic therapy in congenital nasolacrimal duct obstruction.

AIMS: To determine the current bacteriology of mucopurulent discharge in congenital nasolacrimal duct obstruction (CNDO), the in vitro response to different antibiotics and clinical effectiveness of the antibiotics used to relieve babies from mucopurulent discharge. METHODS: A clinical study evaluated the effectiveness of local antibiotic agents clinically and in vitro. 50 samples were obtained from the lacrimal sac in 47 young children with CNDO. The patients' mean age was 21.45 +/- 17.09 months. The cultures were incubated and the infectious agents isolated. Sensitivity testing was performed in each case, testing 10 different local antibiotics. A control group of 10 babies expected for cataract surgery was constituted. RESULTS: Cultures were positive for bacteria from 72.64% of the samples. 73 isolates were recovered from the 50 samples. The bacterial species most frequently cultured was Streptococcus pneumoniae, representing 35.4% of the isolates, followed by Haemophilus influencae (19.6%). The sensitivity testing revealed ofloxacin and tetracycline to be the most effective drugs as monotherapy. Clinically the combination of bacitracin and neomycin, primarily used in half of the patients as initial therapy, was successful in curing the dacryocystitis in 82.5% of all patients. CONCLUSION: Chronic dacryocystitis due to CNDO is associated with an equal proportion of Gram positive and negative bacteria, which can be treated with a high effectiveness by a combination drug of bacitracin and neomycin.

Endoscopic laser recanalisation of presaccal canalicular obstruction.

AIM: To document the results of erbium (Er)-YAG laser treatment in presaccal canalicular obstruction in combination with the use of a flexible endoscope. METHODS: For the first time an Er-YAG laser (Schwind, Sklerostom) was attached to a flexible endoscope (Schwind, Endognost) and used to recanalise a stenosis of the upper, lower, or common canaliculus. In 17 patients (mean age 41.5 (SD 11.9) years), 19 treatments (two bilateral) were performed. In all cases the scar was observed using the endoscope and was excised by laser ablation. A silicone intubation was performed in all cases. In addition to the endoscopy an irrigation was performed to prove the intactness of the lacrimal pathway system after laser treatment. RESULTS: Membranous obstructions with a maximum length of 2.0 mm (14 procedures) in the canaliculus were opened easily using the laser, and the silicone intubation was subsequently performed without difficulty. Scars thicker than 2.0 mm could not be opened safely without canaliculus penetration (five procedures). Irrigation was positive in all cases up to the end of a 6 month period, providing the tubes remained in place. The maximum follow up is now 17 months (minimum 8 months) and in 16 cases (84.2%) the canaliculi are still intact. CONCLUSION: Endoscopic laser treatment combined with silicone intubation enables us to recanalise presaccal stenoses of canaliculi under local anaesthesia up to a scar thickness of 2.0 mm. Best results can be achieved in cases where much tissue can be saved. Under such conditions this procedure can substitute for more invasive surgical techniques, especially a conjunctivo-dacryocystorhinostomy (CDCR).

Long-term results after surgical basal cell carcinoma excision in the eyelid region.

AIMS: To evaluate the data for patients with basal cell carcinoma (BCC) in the eyelid region, to demonstrate histologically controlled tumour excision, and to prove the efficacy of the treatment on the basis of long term observations. METHODS: Retrospective analysis of 382 microscopically controlled BCC excisions in the eyelid apparatus (350 patients) in a follow up study over 5.7 (SD 1.1) years. Tumour location, tumour size, and histological results were recorded. The same procedure was followed for recurrences. Follow up examinations were carried out 1, 3, 6, and 12 months after the operation, and then annually for a further 4 years or longer. RESULTS: A recurrence rate of 5.36% was observed after the primary operation. 60.3% of first recurrences occurred in the medial canthus, 41.2% showed in depth extension, and sclerosing types were overly represented at 35.3%. After the second operation the recurrence rate increased to 14.7% and reached 50% after a third and fourth operation. CONCLUSIONS: The greatest risk of recurrence exists for BCCs of the medial canthus with in depth extension, and for sclerosing types. The recurrence rate increases after every operation. For high risk cases, consideration should be given to adjuvant treatment such as radiotherapy.

[Manifestation of Wegener's granulomatosis as lacrimal sac tumor]. / Manifestation einer Wegenerschen Granulomatose als Tränensacktumor.

HISTORY AND SIGNS: This is a report on a patient, who presented an ulcerated lesion (1.5 x 1.2 cm) in the lacrimal sac area. The lesion had existed for 3 months before the patient went to an ophthalmologist. The lacrimal pathway was rinsable all the time. THERAPY AND OUTCOME: The patient was treated by antibiotics and leukase cones at first. But the lesion showed no tendency to heal under this therapy. The patient was sent to the ETN department for further investigations. At this time the suspicion of a malignant process was discussed. In the meantime the lesion became epithelialized, but no closure could be reached at any time. The CT scans showed a bone defect of 1 cm of the lamina papyracea. Tumorous tissue passed through this defect reaching the ulcer at the medial canthal angle. A biopsy was been taken and revealed granulomatous tissue typical for a Wegener's granulomatosis. The patient was sent to an Internal medicine department, but he died one month later due to systemic disorders of this disease. CONCLUSIONS: Although an acute dacryocystitis is most often the reason for a lacrimal sac swelling and inflammation, in all cases of rinsable lacrimal pathway and long term lacrimal sac ectasy a neoplastic or a granulomatous reason has to be excluded.

[Blepharospasm and acupuncture--initial results of a treatment trial]. / Blepharospasmus und Akupunktur--erste Ergebnisse eines Behandlungsversuches.

Blepharospasm is a dystonia of the orbicularis muscle. The background is still unknown, but it appears together with organic and psychic diseases. The therapy with botulinum toxine is symptomatically but efficient. Because of the relaxing effect on the psychological and autonomous nervous disorders we performed a pilot study with acupuncture. There was a neurological observation first, then a ophthalmologic examination of the ocular surface. Blepharospasm was measured by the Visual Analogue Scale (VAS), the social disorders by the Elston-Score. We performed acupuncture treatment 10 times once weekly. The used points were local points with higher sensitivity, and points with empiric relaxation effect. 5 patients were included. The mean of Elston score was 3 before acupuncture and 4 after acupuncture. The VAS increased from 33.3 to 69/100 points. But there were fluctuations of dystonias. In conclusion acupuncture maybe a good method for this difficult disease additionally.

[Frontalis suspension with "expanded polytetrafluoroethylene (ePTFE) strips" in congenital ptosis]. / Frontalissuspension mittels "expanded- Polytetrafluoroethylene (ePTFE)-Streifen" bei kongenitaler Ptose.

BACKGROUND: The Frontalis Suspension Technique is indicated in cases with minimal or no levator function. At the beginning sutures were used as sling material and after further modifications suture material was replaced by autologous or homologous fascia lata. In the last years ePTFE has proved to be a very suitable sling material. PATIENTS AND METHODS: Since ePTFE is very biocompatible, it was used for this modified frontalis suspension technique. The anterior tarsal surface is exposed and a small tunnel is created between the skin incision and the second incision superior to the brow. An 0.3 mm thin ePTFE strip is induced into the tunnel and connects the upper lid with the frontalis muscle. The ePTFE Soft Tissue Patch must be exactly adapted to the tarsus and has to be deeply sutured to the frontalis muscle below the brow incision. Since 1994 17 modified frontalis suspension procedures have been performed on 14 patients. RESULTS: The functional and cosmetic result were good in nearly all patients. No implant had to be removed during the follow up period. DISCUSSION: The new technique of frontalis suspension using a ePTFE strip guarantees a regular upper lid lifting by the axial and direct connection of the anterior tarsal surface with the frontalis muscle.

[Recent developments in lacrimal duct endoscopy]. / Neuere Entwicklungen der Tränenwegsendoskopie.

BACKGROUND: At the moment digital dacryocystography yields the most exact results in lacrimal diagnostics. The main disadvantage lies in the dependency of high tech X-ray systems and an angiography unit, which raise enormously the costs of the examination. Recently a new diagnostic system of the lacrimal pathway is available. PATIENTS AND METHODS: Miniaturizing of endoscopes and new developments in the fiberoptic technology made it possible to introduce these mini-endoscopes into the canaliculi and perform antegrade examination of canaliculi, lacrimal sac and ductus nasolacrimalis. 86 patients agreed to this examination. With this examination a direct visualisation of the mucous membrane of the lacrimal pathways and possible pathological alterations were possible. RESULTS: In case of axial illumination and syringing of the lacrimal system good visibility can be achieved and an exact examination is possible. Especially for endoscopy modified probes of Bangerter provide an exact examination at narrow areas and areas of bendings too. Different reasons for stenosis of the lacrimal pathways could be identified, especially 2 cases of a lacrimal sac tumor. CONCLUSIONS: The use of this new miniendoscope provides a very good examination under direct visualisation of the lacrimal pathway independent of a high tech X-ray equipment. The goal is to use this new endoscope on the one hand only for diagnostic reasons as endoscope and on the other hand in combination with a laser fiber to perform an antegrade laser dacryocystorhinostomy.

[Congenital toxoplasmosis retinochoroiditis after primary infection of the mother in pregnancy]. / Konnatale Toxoplasmose retinochoroiditis nach Primärinfektion der Mutter in der Schwangerschaft.

An infection with Toxoplasma gondii during pregnancy means that the unborn baby may be infected as well. In addition to the risk that the baby may be born dead, central chorioretinitis could also occur. Pregnant women with primary infection were treated with spiramycin or a combination of pyrimethamine and sulfadiacine. A total of 153 children (1 month-1 year) were examined twice with particular attention to the macula region. Only two cases showed central retinochoroiditis. In one case the treatment was insufficient, and in the other the time between the serological examinations was too long. None of the other children showed any central pathological changes within the follow-up period. Screening methods, the appropriate treatment and the results are presented.

[Monoca silicone intubation of the lacrimal ducts after retrograde dilatation]. / Monoka-Silikonintubation der Tränenwege nach retrograder Dilatation.

The causes of stenosis in the area of the nasolacrimal duct in adults are extremely variable. In general, the symptoms may vary, but most cases exhibit a common factor of circumstances in the prestenotic area which favor recurring inflammation. The treatment of these disorders is limited to either a conservative therapeutic approach to control inflammation, or surgically invasive measures. By using balloon catheters, which are usually applied in PTCA (percutaneous transluminal coronary angioplasty), dilatation of the relative postsaccal stenosis is obtained with the assistance of radiography. An exact diagnosis by means of various testing methods, including digital dacryocystography for the detailed localization and documentation of any pathological changes, is decisive for success. Only in cases of incomplete, postsaccal stenosis is retrograde balloon dilation of the distal nasolacrimal duct incated. The guide wire, designed for the PTCA balloon catheter set, is introduced via the canaliculus to the nasal cavity antegradely and caught with a thin hook and pulled from the naris. This is done under visual control using an image converter. The balloon catheter is retrogradely threaded over the guide wire. The balloon is then placed at the site of the pathological stenosis under X-ray control and dilated with high pressure. To assure permeability of the system, a monocanalicular silicone intubation has to be made immediately following this procedure. This procedure has been performed successfully on six patients; Follow-up time ranged from 6 to 22 months. The initial results are encouraging and sustain hope that this minimally invasive, interdisciplinary technique represents a new alternative in the treatment of incomplete, postsaccal lacrimal stenosis.

Retrograde dilation of postsaccal lacrimal stenosis.

The causes of nasolacrimal duct stenosis in adults can vary greatly. In general, the symptoms can also vary, but most cases share a tendency toward recurring inflammations in the prestenotic area. The treatment of these disorders is limited to either conservative therapy to control inflammation or surgically invasive measures. By using balloon catheters, usually applied in percutaneous transluminal coronary angioplasty (PTCA), dilation of the relative postsaccal stenosis can be performed under radiographic control. An exact diagnosis using various testing methods, including digital dacryocystography for detailed localization and documentation of any pathologic changes, is decisive to success. Only in cases of incomplete postsaccal stenosis is retrograde balloon dilation of the distal nasolacrimal duct indicated. A guide wire, designed for the PTCA balloon catheter set, is introduced via the canaliculus to the nasal cavity antegradely and caught with a thin hook and pulled from the naris, under visual control with an image converter. The balloon catheter is retrogradely threaded over the guide wire. The baloon is then placed at the site of the pathologic stenosis under radiographic control and dilated with high pressure. To ensure the permeability of the system, monocanalicular silicone intubation has to be performed immediately afterwards. This procedure has been performed successfully on 6 patients with a follow-up of 6 to 27 months. These initial results give rise to the hope that this minimally invasive, interdisciplinary technique represents a new alternative in the treatment of incomplete postsaccal lacrimal stenosis.

Interaction of carbonyl cyanide 3-chlorophenylhydrazone with cytochrome c oxidase.

Cytochrome c oxidase binds protonophore carbonyl cyanide 3-chlorophenylhydrazone (CCCP) with high affinity. There are 1.46 high-affinity binding sites per cytochrome c oxidase for CCCP with dissociation constant 2.7 x 10(-7) mol/l. The bond between the CCCP and cytochrome c oxidase accomplishes through the group on cytochrome c oxidase with pKa 6.64 and is based on the electrostatic interaction. Interaction of CCCP with low-affinity binding sites of cytochrome c oxidase induces the shift of the anion CCCP spectrum to UV-region. The similar effect is characteristic for CCCP interaction with protons. Lipophilic non-dissociated derivative NCH3CCP is not binding to cytochrome c oxidase.

Gore-Tex soft-tissue patch frontalis suspension technique in congenital ptosis and in blepharophimosis-ptosis syndrome.

The frontalis suspension technique is the surgical method of choice in ptosis patients with a levator function of 2 mm or less. This type of ptosis is found unilaterally or bilaterally in isolated congenital ptosis and mostly bilaterally in blepharophimosis-ptosis patients. The frontalis suspension technique is frequently used in children being operated on because of congenital ptosis, where the excision of the fasciae latae can be avoided so as not to risk scarring the donor area. Since the new heterologous material polytetrafluoroethylene (Gore-Tex soft-tissue patch) has been available (1986), it has been used on our ptosis patients with minimal levator function instead of the formerly much used fasciae latae. Thirty-seven frontalis suspension procedures have been performed in 26 patients, of whom 6 were children who suffered from bilateral blepharophimosis-ptosis syndrome. All ptosis procedures were performed as Fox pentagons, and in the combined cases, the medial canthoplasty was carried out as a bilateral one-stage procedure. The postoperative healing was free of complications. Three years postoperatively one patient did exhibit rejection of one of the two implanted Gore-Tex slings. In all other patients, the functional and cosmetic results were satisfying, and high biocompatibility of the Gore-Tex soft-tissue patch material could be shown.

Dynamic documentation of digital dacryocystography.

Digital dacryocystography has proved to be the optimum method of x-ray diagnostics in connection with managing obstructions of the lacrimal pathways. We present 50 patients in whom digital dacryocystography was performed with a computer-controlled x-ray unit and a C-arc coupled to an image intensifier TV-system. A nonionic water soluble contrast medium was used. The bilateral contrast medium filling process appearing on the monitor was simultaneously recorded on videotape. Videocassettes, with patient data attached, were stored in a videotape library, providing repeatable diagnostic documentation and an excellent base for scientific analyses.

[Documentation of digital dacryocystography using video technique]. / Dokumentation der digitalen Dakryozystographie mittels Videotechnik.

Digital dacryocystography is considered the optimal radiodiagnostic method in obstructions of the lacrimal pathways. The examination is carried out by a computer-controlled X-ray unit with a C-arc attached to an image-intensifying TV system. In this way the advantages of this technique and of modern video recording are linked. The bilateral filling process of the contrast medium is recorded and the findings simultaneously recorded on videotape. Consequently, pictures of the dynamic flow are retrievable at any time. This provides good diagnostic documentation and an outstanding basis for any scientific analysis.

Spectrophotometric detection of the interaction between cytochrome c and heparin.

Heparin inhibits transport of electrons from reduced cytochrome c to cytochrome c oxidase. The effect is due to the interaction of heparin with cytochrome c. It has been observed that binding of heparin to the reduced or oxidized cytochrome c changes the spectrum of cytochrome c at the Soret region. Affinity chromatography of heparin in cytochrome c immobilized to thiol-Sepharose shows that commercial heparin is eluted in the low-affinity and high-affinity fractions. Both participate in the interaction with cytochrome c. Polylysine induces decay of the cytochrome c-heparin complex.

Circulating ICAM-1 levels in serum of uveitis patients.

Intercellular Adhesion Molecule-1 (ICAM-1) is a cytokine-inducible adhesion molecule expressed on cells of multiple lineages at sites of inflammation. Recently a truncated form of ICAM-1 has been discovered to be circulating in serum. This study reports on circulating serum (cICAM-1) levels in 132 uveitis patients (HLA-B 27 pos. acute anterior uveitis (AAU); HLA-B27 neg. anterior uveitis (AU); intermediate uveitis (IU); heterochromic cyclitis Fuchs (HCF); sarcoidosis; Toxoplasmosis). Measurement of circulating ICAM-1 serum levels was performed using a monoclonal antibody based ELISA, with healthy blood donors serving as the control group. Applying multiple variance analysis and the Student Newmann-Keuls test we found a statistically significant elevation of serum cICAM-1 level in the HLA-B 27 neg. AU group (n:31), in the IU group (n:25) and in patients with sarcoidosis (n:18). Serum levels of HLA-B27 pos. AAU patients, patients with HCF and patients suffering from ocular toxoplasmosis did not differ significantly from levels of the control group.

Binding of Zn(II) ions to alpha-lactalbumin.

The binding of Zn(II) ions to human and bovine alpha-lactalbumin has been studied by fluorescence, scanning microcalorimetry, and proteolytic digestion. The intrinsic tryptophan fluorescence spectrum of Ca(II)-loaded alpha-lactalbumin is insensitive to Zn(II) binding to the strong cation binding sites (Zn:protein ratios up to 20), yet the thermal denaturation transition, as detected by intrinsic fluorescence, is shifted toward lower temperatures. On the other hand, low concentrations of Zn(II) ([Zn]:[protein] less than 1) shift heat sorption curves toward lower temperatures. It was concluded that alpha-lactalbumin possess several relatively strong Zn(II) binding sites, which are filled sequentially, the process being accompanied by protein aggregation. The strongest Zn(II) binding (5 x 10(5) M-1) increases its susceptibility to tryptic and chymotryptic digestion, slightly decreases its affinity for the fluorescent probe, bis-ANS, and alters its interactions with UDP-galactose. Zn(II) binding to aggregated forms of alpha-lactalbumin increases its affinity to bis-ANS.