bluuebunny/crossref_metadata_2025 huggingface dataset:
Protective Role for bcl-2 in Experimentally Induced Cell Death of Bovine Corneal Endothelial Cells
Authors: Joo Choun-Ki , Cho Kyung-Sun , Kim Hyun-Eui , Choi Jun Sub , Oh Young J.
Published: April 2003

To characterize the pattern of cell death and to investigate the potential role of bcl-2 in a death paradigm of corneal endothelial cells, primary cultures of bovine corneal endothelial (BCEN) cells were first established and treated with 0.01–1 µM staurosporine, a nonspecific protein kinase inhibitor. The pattern of BCEN cell death induced by staurosporine was apoptotic in nature, characterized by shrinkage of the cytoplasmic membrane, nuclear condensation and DNA fragmentation. Cotreatment of BCEN cells with Z-VAD-fmk (a caspase inhibitor) but not cycloheximide (a protein synthesis inhibitor) prevented staurosporine-induced cell death. To investigate the potential role of bcl-2, BCEN cells were transferred with a eukaryotic expression vector containing anti-apoptotic bcl-2 cDNA and characterized by reverse transcription-polymerase chain reaction (RT-PCR; BCEN/bcl-2). As measured by the MTT reduction assay after treatment with staurosporine, the survival rate of BCEN/bcl-2 cells was 48.0 ± 4.8% compared to 7.4 ± 2.1% in control BCEN cells. As determined by light microscopy, apoptotic changes such as nuclear condensation and apoptotic bodies were largely attenuated in BCEN/bcl-2 cells after staurosporine treatment although arborization of processes and rounding up of the cell body were not affected by overexpression of bcl-2. These results suggest that staurosporine induces apoptosis in a cycloheximide-independent but caspase-dependent manner and bcl-2 acts as a negative regulator in staurosporine-induced apoptosis of BCEN cells.
Does Convergence, Not Accommodation, Cause Axial-Length Elongation at Near?
Authors: Bayramlar Hüseyin , Çekiç Osman , Hepşen İbrahim F.
Published: April 2003

To determine whether convergence rather than accommodation has a primary effect on the changes in axial length and other biometric components during near fixation, we measured the anterior chamber depth, lens thickness, vitreous length and axial length in the right eyes of 124 young male subjects while their left eyes focused at distance (6 m) and near (20 cm). The measurements were performed before and after cycloplegia in the right eye, so we aimed to study biometric components of the eye in the states of accommodation and nonaccommodation, but converging at near. While the left eye focused at near, the axial length increased significantly with and without cycloplegia (p < 0.0005 and p < 0.0005). The vitreous length was the main increasing ocular biometric component at near both with and without cycloplegia (p < 0.044 and p = 0.001, respectively). At near, there was no difference between two mean axial length and two vitreous length measurements both with and without cycloplegia (p = 0.672 and p = 0.595, respectively). Under cycloplegia, anterior chamber depth also increased significantly at near fixation (p = 0.012). Axial elongation at near fixation, mainly due to an increase in vitreous length, may result from the effect of accommodative convergence rather than accommodation itself. Much use of convergence, not accommodation, may be one of the contributing factors in adult onset and adult progression of myopia.
Cataract Development in 12-Month-Old Rats Fed a 25% Galactose Diet and Its Relation to Osmotic Stress and Oxidative Damage
Authors: Ohta Yoshiji , Yamasaki Takashi , Niwa Takafumi , Goto Hajime , Majima Yoshinao , Ishigruo Isao
Published: April 2003

We attempted to clarify the pattern of cataract development in 12-month-old rats fed a 25% galactose diet and to assess the relation of cataract development with osmotic stress and oxidative damage. In lenses of 12-month-old male Wistar rats fed a 25% galactose diet over an 8-month period, suture accentuation appeared at 6 months of galactose feeding and then opacities developed from the anterior subcapsular cortex toward the posterior subcapsular cortex, reaching the nuclear region at 8 months of galactose feeding. Increases in lens galactitol and lipid peroxide contents and a decrease in lens reduced glutathione content occurred at 4, 6 and 8 months of galactose feeding. The increase in lens lipid peroxide content and the decrease in lens reduced gluthatione content were accelerated with an increase in feeding period, while the increase in lens galactitol content was decelerated. An increase in lens water content and a decrease in lens protein content occurred at 6 and 8 months of galactose feeding. The lens vitamin E content increased at 6 months of galactose feeding and this increase was concomitant with increases in serum vitamin E and total cholesterol concentrations. The serum lipid peroxide concentration increased at 4 and 6 months of galactose feeding. The present results indicate that in lenses of 12-month-old rats fed a 25% galactose diet, suture accentuation appears initially and then opacities develop from the anterior subcapsular cortex toward the posterior subcapsular cortex, finally reaching the nuclear region. These results also suggest that in the galactosemic aged rats, osmotic stress would mainly contribute to cataract formation, while oxidative damage could be linked to both cataract formation and progression, although an increase in lens vitamin E content occurs during the cataract development.
Steroid Sensitivity and Postoperative Course of Seven Patients with Behçet’s Disease
Authors: Tanaka Takao , Suzuki Jun , Yamakawa Naoyuki , Usui Masahiko
Published: April 2003

Systemic administration of steroids to patients with Behçet’s disease in Japan is often followed by a worsening in ocular symptoms. The sensitivity of 7 patients with Behçet’s disease to steroids which were administered during and after intraocular surgery was therefore measured, and their postoperative course was reviewed. Three of the 7 showed a low steroid sensitivity, and the frequency of postoperative inflammatory episodes and the intraocular pressure were both higher for this group than for the 4 who showed high steroid sensitivity.
Subretinal Injection of Rod Outer Segments Leads to an Increase in the Number of Early-Stage Melanosomes in Retinal Pigment Epithelial Cells
Authors: Peters S. , Kayatz P. , Heimann K. , Schraermeyer U.
Published: April 2003

Our study was performed to test the hypothesis that subretinally injected protein can induce melanogenesis in the retinal pigment epithelium (RPE). Rod outer segments (ROS) were isolated from cattle eyes and injected into the subretinal space of Long Evans rats. Five days after surgery, the injected eyes were investigated by electron microscopy. The number of early-stage melanosomes and small melanin granules was compared in injected and noninjected eyes. It was found that the injected ROS were phagocytized by the RPE cells, and the number of early-stage melanosomes in the RPE was significantly increased in injected eyes compared to eyes without injection. The ROS-containing endosomes fused with melanolysosomes in which melanogenesis took place. The increased number of early-stage melanosomes indicates new formation of melanin.
Evaluation of the Circulation in the Retina, Peripapillary Choroid and Optic Disk in Normal-Tension Glaucoma
Authors: Sugiyama Tetsuya , Schwartz Bernard , Takamoto Takenori , Azuma Ikuo
Published: April 2003

Computerized image analysis, including fluorescein angiography, was used to evaluate the retinal, choroidal and optic disk blood flow in 16 patients with normal-tension glaucoma (NTG) and to correlate this measurement with visual fields, retinal vessel width, optic disk pallor and blood pressure (BP). The angle of the ascending slope of the fluorescein dye curve was measured as an index of blood flow from the densitometric and time curves of the fluorescein angiograms in the optic disk, peripapillary choroid, retinal artery and vein for each quadrant. While the ascending slope as well as the retinal vessel width were most reduced in the inferior and nasal regions, the mean threshold was lowest in the superior and nasal quadrants. There were positive significant correlations between artery width and threshold value, between angles of slopes and pallor. In addition, systolic BP had a negative correlation with pallor, and diastolic BP had a positive one with slope in the choroid. These results indicated the probable association of a decrease in retinal, choroidal and optic disk blood flow in the inferior and nasal quadrants as well as vessel width in the inferior nasal quadrant with visual field loss in the superior quadrant, and also demonstrated an increasing blood flow for enlargement of pallor. A decrease in BP was found to be related to reduced blood flow in choroid and optic disk impairment.
Linoleic Acid Cytotoxicity to Bovine Lens Epithelial Cells: Influence of Albumin on Linoleic Acid Uptake and Cytotoxicity
Authors: Trimborn Michael , Iwig Martin , Glanz Dagobert , Gruner Michael , Glaesser Dietmar
Published: April 2003

The high cytotoxicity of linoleic acid (LA) to cultured bovine lens epithelial cells is correlated with high uptake rates for the fatty acid (FA). Both, LA uptake and LA cytotoxicity strongly increase with the increasing LA-to-albumin molar ratio in the culture medium. Cellular uptake and cytotoxicity of LA can be competitively inhibited with the noncytotoxic palmitic acid. The findings may be of interest in view of the low albumin concentration in aqueous humor, resulting in extremely low buffering capacities for free FAs including LA, oleic acid and other cytotoxic cis-unsaturated free FAs, which are strongly raised in pathological situations like diabetes mellitus.
Effect of Rabbit Aqueous Humor Obtained after Cataract Surgery on Collagen Gel Contraction Induced by Bovine Lens Epithelial Cells
Authors: Kurosaka Daijiro , Kato Katsuhiko , Oshima Takeshi , Kurosaka Hiroyo , Yoshino Mami , Nagamoto Toshiyuki
Published: April 2003

We evaluated the effect of specimens of pre- and postoperative aqueous humor on the contraction of collagen gels, and the effect of transforming growth factor-β2 (TGF-β2) in postoperative aqueous humor. Rabbit aqueous humor was collected preoperatively and on postoperative day 7. Bovine lens epithelial cells (LECs) were cultured in collagen gel in F-12 nutrient mixture supplemented with 5% fetal bovine serum that contained 10% aqueous humor obtained under various conditions. Gel area was determined on day 4. Gels cultured with the medium that contained phosphate-buffered saline showed a statistically significant contraction after 4 days. Aqueous humor from aphakic or pseudophakic eyes significantly increased contraction, with both specimens having a similar effect. Approximately 60% of the contractile effect of the postoperative aqueous humor was neutralized by anti-TGF-β2 antibody. However, the promoting effect of the aqueous humor sampled postoperatively was less than that sampled preoperatively. Although the aqueous humor obtained postoperatively increased the contractility of the LECs, with the level of TGF-β2 apparently responsible for much of its effect, the effect was less than that observed in the aqueous humor obtained preoperatively.
Plasma Membrane Calcium ATPase Gene Expression in Bovine Lens Epithelium
Authors: Bian Lijun , Zeng Junwen , Borchman Douglas , Paterson Christopher A.
Published: April 2003

Plasma membrane calcium adenosine triphosphatase (Ca2+-ATPase) is an energy-dependent protein responsible for transporting cytosolic calcium across the plasma membrane. Multiple plasma membrane Ca2+-ATPase isoforms are expressed from four genes (PMCA1–4) and alternative mRNA splicing. We have studied PMCA gene expression in bovine lens epithelium tissues by reverse transcription-polymerase chain reaction, Southern blot, and Northern blot hybridization. All four PMCA genes are expressed in the lens epithelium, the PMCA3 transcript being the most abundant. The transcripts for PMCA1, PMCA2, and PMCA4 exist in decreasing order of abundance. There is no evidence for the expression of any novel PMCA genes in bovine lens epithelium.
Expanded Polytetrafluoroethylene Episcleral Implants Used as Encircling Scleral Buckling
Authors: Korobelnik J.F. , D’Hermies F. , Chauvaud D. , Legeais J.M. , Hoang-Xuan T. , Renard G.
Published: April 2003

To investigate tissue changes induced by the implantation of expanded polytetrafluoroethylene (e-PTFE) used as encircling scleral buckling, an experimental and histopathological study was performed in rabbits. The eyes of 12 rabbits were implanted during 20–128 days with e-PTFE episcleral implants. Of these, 8 eyes were uncomplicated and enabled us to perform the histopathological study, 2 others were discarded for too short a time of implantation and, in 2 additional eyes, the episcleral implant was exposed with a wide conjunctival erosion. Surrounding the implant, a constant encapsulation was combined with numerous giant cells, forming a granuloma caused by irregularities of the implant outline. Its porous structure allowed a peripheral colonization by fibrovascular tissue. Scleral thinning and scleral invagination were the main scleral changes. No change of the conjunctiva made us suspect that a granuloma occurred before it was found histologically. The granulomatous changes did not seem to modify the apparently good experimental tolerance of the material after its implantation.
Epidemiological Study of Nontraumatic Phakic Rhegmatogenous Retinal Detachment
Authors: Ivanišević Milan , Bojić Lovro , Eterović Davor
Published: April 2003

We studied the epidemiological characteristics of nontraumatic phakic rhegmatogenous retinal detachment (RRD) in a defined population of Split-Dalmatia County, Croatia. A total of 278 eyes of 272 patients developed RRD during an 11-year period, 1988–1998, with a population of 465,947. The annual incidence was 5.4 per 100,000 population. The mean age of patients was 58.3 years, and the sex distribution corresponded with that in the general population. There was insignificant preponderance of right eye involvement. Bilaterality was observed in 2.2%. The presence of myopia was diagnosed in 46.9% eyes.
Tear Nitrite and Nitrate Levels as Nitric Oxide End Products in Patients with Behçet’s Disease and Non-Behçet’s Uveitis
Authors: Mirza G. Ertuğrul , Karaküçük Sarper , Er Mustafa , Güngörmüş Naci , Karaküçük İnci , Saraymen Recep
Published: April 2003

To evaluate the role of nitric oxide (NO) in ocular inflammation, tear nitrite and nitrates (NN) as NO end products were determined in 11 patients with Behçet’s disease (BD) and in 11 with non-Behçet’s uveitis (NBU) during the active and remission stages and in 12 healthy controls. Median (with the range) NN levels were 82.29 (59.60–98.25) µmol/l in the active and 98.25 (52.88–246.92) µmol/l in the remission stage of BD; 88.17 (25.99–116.73) µmol/l in the active and 83.00 (31.04–250.28) µmol/l in the remission stage of NBU and 109.17 (88.17–158.74) µmol/l in the controls. The NN levels in the active stage of BD and NBU were significantly decreased when compared to the controls (p < 0.05; Kruskal-Wallis test). Decreased NN levels at the activation stage may be caused by the rapid transformation of the NO to peroxynitrites, which are highly oxidizing and cytotoxic substances.
Retinal Cells Produce TIMP-1 and TIMP-2 in Response to Cyclic Mechanical Stretching
Authors: Namba Masako , Matsuo Toshihiko , Shiraga Fumio , Ohtsuki Hiroshi
Published: April 2003

Purpose: Tissue inhibitors of metalloproteinases (TIMPs) are crucial for the maintenance of retinal extracellular matrix such as interphotoreceptor matrix and internal limiting membrane. This study is to examine whether retinal cells respond to mechanical stretching and produce TIMPs. Methods: Chick retinal adherent cells in near confluency were exposed to mechanical stretching of the bottom of a 6-cm Petri dish at the maximum magnitude of 4,500 microstrain and at a cycle of 30 s for 72 h. TIMP-1 and TIMP-2 levels in the medium at 24, 48 and 72 h after the beginning of stretching were measured by enzyme immunoassay, and their expression was examined by immunohistochemistry. Results: The number of retinal cells during the 72-hour period of stretching did not change significantly both in the stretched group and in the nonstretched control group. Retinal cells in the stretched group produced significantly larger amounts of TIMP-1 and TIMP-2 at 48 h after stretching, compared with nonstretched controls (p = 0.0163 and p = 0.047, respectively, Mann-Whitney U test). Immunohistochemically, a large part of retinal cells in nonstretched Petri dishes was positive for glial fibrillary acidic protein, indicative of glial cells, while some small foci of cells were positive for neuron-specific enolase, indicative of neurons. Fluorescent double labeling demonstrated that both glial cells and neurons were positive for TIMP-1 and TIMP-2. Conclusion: Chick retinal cells, most of which were glial cells mixed with a small number of neurons, produced TIMP-1 and TIMP-2. Their production was enhanced by cyclic mechanical stretching.
The Expression of Multiple Cytokines and Inducible Nitric Oxide Synthase in Experimental Melanin-Protein-Induced Uveitis
Authors: Kim Hae-Sook , Yoon Sungjoo Kim , Joo Choun-Ki
Published: April 2003

We investigated the kinetics of multiple cytokines and inducible nitric oxide synthase (iNOS) in experimental uveitis induced by bovine melanin protein (BMP) for the proper treatment of uveitis. Experimental uveitis was induced in male Lewis rats by injection of BMP. The levels of various inflammatory cytokines and iNOS mRNAs were semiquantified by the reverse-transcriptase reaction followed by PCR. The uveitis was started to develop at approximately day 14 and peaked around 21 days after immunization. The signs of uveitis disappeared by 4 weeks after immunization. When the inflammation was severest, TNF-α, IL-1α, IL-10, IFN-γ and iNOS mRNA increased to their peak, which varied with the degree of induction and different time course. We concluded that both cytokines and iNOS might modulate the inflammation at different states of experimental melanin-protein-induced uveitis. Their combination will be necessary for an effective treatment of inflammation.
Tegmental Defects and Cerebrospinal Fluid Otorrhea
Authors: Valtonen Hannu J. , Geyer Carl A. , Tarlov Edward C. , Heilman Carl B. , Poe Dennis S.
Published: April 2003

Congenital tegmental defects that present as unsuspected cerebrospinal fluid (CSF) otorrhea are diagnostic and therapeutic challenges. We reviewed 5 such patients to determine an optimal strategy for evaluation. Five patients presented with watery otorrhea, 4 of them after ventilation tube placement, and only 1 with rhinorrhea. The preoperative analysis of middle ear effusion for β2-transferrin was positive in 2/4, equivocal in 1/4 and false negative in 1/4. Computerized tomography (CT) revealed nonspecific tegmental defects in all 5 patients. Magnetic resonance imaging (MRI) demonstrated meningoencephalocele in 3/5 and dural irregularity in 1/5. Tegmental defects were confirmed at surgery in all cases, demonstrating meningocele or arachnoid granulations in 2/5 and encephalocele in 2/5 patients. We recommend a combination of β2-transferrin analysis to verify CSF, high resolution CT (axial and coronal planes) to diagnose tegmental defects, and MRI (multiplanar) to evaluate the type of herniation. A combination mastoid and middle fossa approach for definitive repair is suggested.
Possibility of Reciprocal Infection of Methicillin-Resistant Staphylococcus aureus between Medical Personnel and Patients Undergoing Middle Ear Surgery
Authors: Lee Eun-Soo , Song Jong-Seok , Hwang Soon-Jae , Suh Han-Kyu , Cheong Hee-Jin
Published: April 2003

The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection after middle ear surgery has recently increased in our hospital. In this study we tried to determine whether the strains of MRSA isolated from infected patients are identical to those obtained from medical personnel, to prove a reciprocal transmission between medical personnel and patients. Surveillance bacterial cultures of medical personnel were performed from the anterior nares and from the fingertip. Molecular epidemiological studies, ribotyping and pulsed-field gel electrophoresis (PFGE) were used to compare the 12 MRSA strains obtained from carriers among medical personnel with the 60 strains identified from patient’s otorrhea. Six different MRSA strains were identified from ribotyping, and three subtypes from PFGE. There was a particular subtype which was the most frequently identified strain found in both medical carriers and patients. Postoperative MRSA infection rates after the treatment of medical carriers and application of preventive procedures decreased from 11.9 to 5.7%. These findings suggest that the MRSA transmissions have occurred between medical personnel and patients.
Dislocation of the Incus into the External Auditory Canal after Mountain-Biking Accident
Authors: Saito Takehisa , Kono Yoko , Fukuoka Yoko , Yamamoto Hideyuki , Saito Hitoshi
Published: April 2003

We report a rare case of incus dislocation to the external auditory canal after a mountain-biking accident. Otoscopy showed ossicular protrusion in the upper part of the left external auditory canal. CT indicated the disappearance of the incus, and an incus-like bone was found in the left external auditory canal. There was another bony and board-like structure in the attic. During the surgery, a square-shaped bony plate (1 × 1 cm) was found in the attic. It was determined that the bony plate had fallen from the tegmen of the attic. The fracture line in the posterosuperior auditory canal extending to the fossa incudis was identified. According to these findings, it was considered that the incus was pushed into the external auditory canal by the impact of skull injury through the fractured posterosuperior auditory canal, which opened widely enough for incus dislocation.
Expression of Inducible Nitric Oxide Synthase in the Cochlea following Immune Response in the Endolymphatic Sac of Guinea Pigs
Authors: Watanabe Ken-ichi , Tomiyama Shunichi , Jinnouchi Ken , Pawankar Ruby , Yagi Toshiaki
Published: April 2003

Immunohistochemical study for inducible nitric oxide synthase (iNOS or NOS II) in the cochlea of guinea pigs was performed after the injection of keyhole limpet hemocyanin (KLH) into the endolymphatic sac. Morphological changes were observed in the cochlea of all animals after the injection of KLH. Increased iNOS expression was detected in the lateral wall, organ of Corti and ganglion cells. It is known that high levels of nitric oxide can lead to inner ear dysfunction. Our results suggest that iNOS may mediate the inner ear disturbance as seen in endolymphatic hydrops.
Botryoid Rhabdomyosarcoma of the Nose: Pitfalls of Pathology
Authors: Caversaccio Marco , Stauffer Edouard
Published: April 2003

A 2-year-old Swiss boy was referred to our hospital because of an obstructive sleep apnea syndrome with suspected recurrent choanal polyp. During surgery, a polypoid soft mass was found in the right nose and extirpated. The histology of the resected polypoid tumor seemed compatible with a polyp; however, immunohistochemistry revealed a botryoid rhabdomyosarcoma. Rhabdomyosarcomas are rare. The most important differential diagnoses for rhabdomyosarcoma are discussed and methods available for differentiating are included.
General Considerations on Distant Metastases from Head and Neck Cancer
Authors: Ferlito Alfio , Rinaldo Alessandra , Buckley J. Graham , Mondin Vanni
Published: April 2003

Mortality in head and neck cancer is due to locoregional disease, distant metastases or intercurrent disease. As treatment of the primary tumor and cervical metastases has improved, the proportion of deaths from co-morbidity and from distant metastases has increased. Distant metastases almost invariably herald a poor prognosis in head and neck cancer with an average survival of 4.3–7.3 months and treatment is usually palliative. Reliable detection is important to prevent inappropriate treatment. The risk is related to the site, stage and histology of the primary tumor and the presence of cervical metastases. Early detection and treatment of cervical metastases may prevent distant metastases. Accurate staging of tumors helps to identify high-risk tumors that should be specifically investigated for distant metastases.
Distant Metastases from Sinonasal Cancer
Authors: Lund Valerie J.
Published: April 2003

Distant metastases from sinonasal malignancy are generally a rare event except in the terminal stages of the diseases and many patients die from recurrence before the secondaries become clinically manifest. Sinonasal neoplasia covers a diverse range of pathologies, some of which have a greater tendency to spread than others, in particular adenoid cystic carcinoma, malignant melanoma and some of the sarcomas. Notwithstanding this, the frequency with which systemic metastases occurs is such that screening at presentation could not be regarded as cost-effective and is consequently only instituted in the presence of specific symptoms.
The Treatment of Distant Metastases in Head and Neck Cancer – Present and Future
Authors: Buckley J. Graham , Ferlito Alfio , Shaha Ashok R. , Rinaldo Alessandra
Published: April 2003

At the present time the occurrence of distant metastases in patients with head and neck squamous cell carcinoma means that lifespan is measured in months. In most instances treatment is purely palliative. Isolated lung metastasis can be successfully removed with long-term disease control in selected patients. Radiotherapy can be useful for palliation of bone metastases and occasionally lung or brain metastases. Chemotherapy does not have a major impact at the present time except for the treatment of metastases from nasopharyngeal cancer. Palliative symptomatic care, along with appropriate pain control, is essential since pain management is very important in these patients. A significant change in the survival of patients with head and neck cancer is only likely to occur by the development of new approaches to treatment. Blocking tumor angiogenesis and treatment based on genetic abnormalities or cell surface receptors offer the two strategies that are most likely to be successful.
Allopurinol Attenuates Endolymphatic Hydrops in the Guinea Pig Cochlea
Authors: Shinomori Yusuke , Kimura Robert S.
Published: April 2003

Based on the hypothesis that Ca2+ overload in the scala media may produce endolymphatic hydrops and generate free oxygen radicals (FOR), allopurinol, a xanthine oxidase inhibitor and free radical scavenger, was administered to guinea pigs after the surgical obliteration of the endolymphatic duct. Allopurinol was given intraperitoneally (50 mg/kg/day) for 15 days starting 1 day prior to the surgical blockage procedure. Measurements from histological serial sections of these temporal bones showed that the total volume of the scala media was significantly reduced (p = 0.007) compared with control hydropic ears. There was an indication of reduced incidence of atrophy in sensorineural structures and stria vascularis. These findings suggest that allopurinol may attenuate the development of endolymphatic hydrops and cell damage by preventing the formation of FOR or scavenging FOR. This study may lead to a new aspect of treatment for Menière’s disease.
Correlation between MRI Findings and Second-Look Operation in Cholesteatoma Surgery
Authors: Kimitsuki T. , Suda Y. , Kawano H. , Tono T. , Komune S.
Published: April 2003

Two-staged intact canal wall tympanoplasty is a common operation for treatment of middle ear cholesteatoma. MRI provides better tissue differentiation of the middle ear and/or mastoid, which often become occupied with soft density tissue after the first operation. If MRI was able to detect the presence of a recurrent or residual cholesteatoma with sufficient sensitivity and specificity, this may facilitate a decrease in the number of second-look procedures. This study compared MRI findings to surgical findings at second-look surgery and calculated the correlation rates between the two sets of findings. Thirty ears having undergone intact canal wall tympanoplasty for cholesteatoma at the initial operation were examined by MRI prior to the second look. Otoscopic findings of the tympanic membrane were nonsuspect in all cases. The true positive rate was 11/30 (37%) and the true negative rate was 10/30 (33%), leading to a radiosurgical correlation of 70%, whereas the false positive rate was 6/30 (20%) and the false negative rate was 3/30 (10%). This indicates that 30% of the MRI findings were incorrect. Therefore, at the present time, MRI does not appear as a likely replacement for second-look surgery in cases of intact canal wall tympanoplasty.
Vertigo and Electronystagmography in Uni- and Bilateral Ménière’s Disease
Authors: Mateijsen D.J.M. , Hengel P.W.J. , Kingma H. , Oreel M.A. , Wit H.P. , Albers F.W.J.
Published: April 2003

In this study we performed standardized measurements of spontaneous nystagmus (SN), caloric tests and velocity step tests on a well-defined group of 92 Ménière patients as part of a diagnostic protocol. The aim of the study was to look for results that could contribute to the diagnosis, but more specifically to a possible classification, of Ménière’s disease. Relations with average hearing loss, shape of the audiogram, average hearing loss, duration and severity of perceived symptoms of disease and differences between uni-and bilateral Ménière’s disease were studied. Directional preponderance (DP) in unilaterally affected patients and labyrinthine preponderance (LP) in both uni- and bilaterally affected patients were significantly more often directed towards the unaffected (uni) or least affected (bi) side (p = 0.007, p = 0.001, p = 0.002, respectively). DP was correlated with LP (uni: R = 0.615, p < 0.001; bi: R = 0.438, p < 0.01), which means that the DP can be seen as a latent SN, directed to the unaffected side. With the caloric test parameters LP and DP it was possible to find unilateral weakness. In unilateral disease a duration of disease <2 years resulted in small asymmetry in time constant (Tasym), large LPs and LPs not only directed to the unaffected, but also to the affected side, whereas a longer duration of disease gave large Tasym and small LPs that were only directed to the unaffected side. Patients suffered more severe vertigo in the first two years of the disease. Studying the shape of the audiogram, it was found that patients having an LP to the affected side, suggesting that this was a hyperactive labyrinth belonging to a beginning of the disease, never had a flat audiogram. In this study the LP was the best diagnostic parameter to find unilateral weakness in patients with Ménière’s disease. Together with the duration of the disease, the severity of symptoms, the asymmetry in T and the shape of the audiogram, it was possible to find two subgroups in the group of patients with unilateral Ménière’s disease. Vestibular data in association with other test results of the diagnostic protocol may contribute to a more sophisticated classification of Ménière’s disease.
Intraoperative Identification of Regenerated Chorda tympani Nerve and Its Relationship to Recovered Taste Function
Authors: Saito Takehisa , Shibamori Yoshiyuki , Manabe Yasuhiro , Yamagishi Takahiro , Igawa Hideki , Yamamoto Takehito , Ohtsubo Toshio , Saito Hitoshi
Published: April 2003

This study demonstrated a simple method of repairing the severed chorda tympani nerve and a method of intraoperative identification of regenerated nerves, and evaluated taste function of regenerated nerves. Seven patients who underwent staged tympanoplasty and whose chorda tympani nerve was severed during primary surgery were evaluated. When the chorda tympani nerve was severed during primary surgery, proximal and distal stumps were anastomosed or approximated almost in the original position and fixed with fibrin glue on the temporal muscle fascia used to reconstruct the eardrum by the underlay method. During primary surgery, end-to-end anastomosis was possible in 3 patients but nerve gap defects remained in the other 4 patients. In all 7 patients, regenerated nerves were identified during secondary surgery not in the tympanic cavity but in the submucosal layer of the previously reconstructed eardrum. In all patients, complete or incomplete recovery of taste perception was observed by both the filter paper disk method and electrogustometry, suggesting that the regenerated nerves had actual taste function. From these results, it was concluded that the severed chorda tympani nerve could regenerate in the reconstructed eardrum even if nerve gap defects remained between the proximal and distal cut ends, when repair or approximation of the nerve was properly completed.
Minimally Invasive Application of Botulinum Toxin Type A in Nasal Hypersecretion
Authors: Rohrbach Saskia , Laskawi Rainer
Published: April 2003

We report on the effect of the local application of botulinum toxin A on nasal hypersecretion in a female patient with intrinsic rhinitis. 20 units of botulinum toxin A (Botox®) was inserted into each nostril using a small sponge in close contact with the lower and middle turbinates. The effect was scored by the patient and by rhinomanometry. Nasal hypersecretion diminished clearly 5 days after the treatment. The rhinomanometric flow increased 2 weeks after the application. No side effects occurred. We conclude that this minimal invasive method of local botulinum toxin application might be a very effective and safe option for the treatment of nasal hypersecretion of different etiologies.
Protein Kinase C-Mediated in vitro Invasion of Human Glioma Cells through Extracellular-Signal-Regulated Kinase and Ornithine Decarboxylase
Authors: da Rocha Adriana B. , Mans Dennis R.A. , Lenz Guido , Fernandes Andréia K. , de Lima Cleber , Monteiro Vanessa F. , Gonçalves Daniela , Moreira José Claudio F. , Brunetto Algemir L. , Rodnight Richard , Schwartsmann Gilberto
Published: April 2003

We investigated the involvement of protein kinase C (PKC) in the in vitro invasiveness of the A-172, U-87 and U-373 human glioma cell lines, as well as the role of ornithine decarboxylase (ODC) and/or extracellular-signal-regulated kinase (ERK) in the actions of PKC. Thus, cells were treated under serum-free conditions with the PKC activator phorbol 12-myristate 13-acetate (PMA), or with the PKC inhibitors bisindolylmaleimide I (GF 109203X) or calphostin C in the absence or presence of the ODC inhibitor D,L-α-difluoromethylornithine (DFMO), and/or the mitogen-activated protein kinase/extracellular-signal-regulated kinase inhibitor 2′-amino-3′-methoxyflavone (PD 098059). Subsequently, cells were assessed for membrane-type 1 matrix metalloproteinase (MT1-MMP) mRNA contents, 72-kD latent, and 59/62-kD activated matrix metalloproteinase 2 (MMP-2) in conditioned media, as well as invasiveness. For these purposes, we used Northern blot analysis, gelatine zymography, and an in vitro filter invasion assay, respectively. Data were related to those found with untreated cells. PKC activity was 2- to 3-fold stimulated by PMA (100 nM for 30 min), and about 2-fold inhibited by calphostin C (40 nM for 2 h) or GF 109203X (5 µM for 20 min). This was accompanied by a similar increase or decrease, respectively, in MT1-MMP mRNA expression, 59/62-kD MMP-2 activity, and in vitro invasion. Inhibition of ODC activity (about 2-fold by 24 h DFMO 5 mM), ERK activation (almost completely by 20 min PD 098059 50 µM), or both these enzymes simultaneously led to a reduction by about half in levels of MT1-MMP mRNA, 59/62-kD MMP-2 activity, and invasion in untreated as well as PMA-stimulated cells. The use of these compounds did not significantly alter the inhibitory effects of GF 109203X or calphostin C. Modulation of PKC and/or ERK activity resulted in corresponding changes in ERK and/or ODC activities, but interference with ODC affected neither ERK nor PKC. Our data suggest a regulatory role for PKC, in co-operation with ERK and ODC, in glioma cell invasion, by modulation of MT1-MMP mRNA expression and MMP-2 activation.
Cyclin D1, Retinoblastoma, p53, and Her2/neu Protein Expression in Preinvasive Breast Pathologies: Correlation with Vascularity
Authors: Heffelfinger Sue C. , Yassin Rawia , Miller Mary Ann , Lower Elyse E.
Published: April 2003

Objectives: Preinvasive breast pathologies show a degree of vascularization that correlates with risk of invasion. Recently, numerous oncogenes and tumor suppressor genes have been shown to regulate neovascularization. Therefore, we examined archival tissues of preinvasive breast pathologies by immunohistochemistry for alterations in the expression of four proteins, cyclin D1, retinoblastoma (Rb), p53, and Her2/neu, known to be important in breast tumorgenesis, and correlated these data with tissue vascularity. Methods: Vascularity was determined by immunologic detection of von Willebrand factor. For carcinoma in situ (CIS) both stromal vascularity (MVD) and vascular cuffing (MCD) were determined. Results: We found that cyclin D1 expression was increased in usual hyperplasia (11% of cases). Atypical hyperplasia, noncomedo CIS and comedo CIS were positive in 43, 49, and 57% of cases, respectively. Changes in Rb and p53 were rare in hyperplasia but occurred in 8 and 10% of CIS, respectively. Her2/neu protein was identified rarely in atypical hyperplasia and in both noncomedo and comedo ductal CIS. Neither Rb nor Her2/neu expression correlated with vascularity. p53 immunoreactivity correlated positively with both MCD and MVD. Cyclin D1 was negatively associated with MVD. Conclusion: These data suggest that p53 and cyclin D1 proteins may regulate the microvessel density of preinvasive breast pathologies.
DNA Topoisomerase II-Alpha Immunoreactivity as a Marker of Tumor Aggressiveness in Invasive Breast Cancer
Authors: Nakopoulou Lydia , Lazaris Andreas C. , Kavantzas Nikolaos , Alexandrou Paraskevi , Athanassiadou Pauline , Keramopoulos Antonios , Davaris Panayiotis
Published: April 2003

Objective: The nuclear enzyme DNA topoisomerase (topo) II breaks and rejoins DNA strands; its isoform topo IIα is associated with active cell proliferation of mammalian cells. The aim of this study was to examine the relationship between the expression of topo IIα and biological behavior markers in breast cancer. Methods: Formalin-fixed, paraffin-embedded tissue from 88 samples of infiltrating breast cancer was immunohistochemically stained for topo IIα. For each case, a topo IIα index was determined by image analysis. Similar indexes were available for Ki-67 protein, a known cell proliferation marker, and p53, bcl-2 and c-erbB-2 oncoproteins. Each case had been staged and graded and the patients had been followed up for a mean period of 61.62 months. Results: Elevated topo IIα immunopositivity (in >10% of malignant nuclei) was detected in 22 tumors, and this immunostatus was statistically associated with poor nuclear differentiation, absence of steroid hormone receptors, high Ki-67 immunoexpression, p53 protein accumulation and c-erbB-2 protein overexpression. Topo IIα expression was not linked with disease extent (stage or lymph node status). Neither proliferation marker (topo IIα or Ki-67) had any significant influence on the patients’ recurrence-free survival. Conclusion: From the above results, we conclude that topo IIα overexpression appears to be linked with cellular dedifferentiation and potentially aggressive tumor phenotype in invasive breast cancer.
Interleukin-1β Induces Different Gene Expression of Stromelysin, Aggrecan and Tumor-Necrosis-Factor-Stimulated Gene 6 in Human Osteoarthritic Chondrocytes in vitro
Authors: Stöve Johannes , Huch Klaus , Günther Klaus-Peter , Scharf Hanns-Peter
Published: April 2003

Objective: To analyze the gene expression of osteoarthritic chondrocytes cultured in alginate after stimulation with interleukin (IL)-1β. Methods: Chondrocytes were isolated from osteoarthritic cartilage obtained during total knee replacement by sequential enzymatic digestion. After suspension in alginate, cells were cultured with and without 100 pg/ml IL-1β. Quantitative RT-PCR reaction was used to estimate the messenger RNA (mRNA) of three different metabolites [tumor-necrosis-factor-stimulated gene 6 (TSG-6), stromelysin-1 (MMP-3) and aggrecan (AGG)]. Results:  After having shown the precision of quantitative PCR, this method allowed us to detect IL-1β-induced changes in mRNA of TSG-6, MMP-3 and AGG. MMP-3 was found to be the most abundant transcript, IL-1β induced a 12-fold upregulation of MMP-3 levels compared to control, and 7-fold of TSG-6. The AGG transcript level, indicating anabolic events, was found to be downregulated by between 2- and 3-fold. Conclusions: In our culture system, the response of osteoarthritic chondrocytes to IL-1β is preserved. Therefore, this system might be helpful for further investigation of the influences of drugs, cytokines and growth factors, for example, on the metabolism of chondrocytes at the level of gene transcription as the most basic level of regulation.
Cell Type-Specific mRNA Quantitation in Non-Neoplastic Tissues after Laser-Assisted Cell Picking
Authors: Bohle Rainer M. , Hartmann Elena , Kinfe Thomas , Ermert Leander , Seeger Werner , Fink Ludger
Published: April 2003

Objective: Cell type-specific mRNA quantitation can be reliably performed after harvesting less than 20 cell profiles from haemalaun-stained cryosections by laser-assisted cell picking. Up to now it has been unclear to what extent these techniques can be used to analyze differential gene expression in complex tissues. Methods: Using a rat model of experimental endotoxin priming of the lung various pulmonary cell types were microdissected from isolated perfused and ventilated rat lungs after aerosol lipopolysaccharide/interferon-γ stimulation. Results: Porphobilinogen deaminase housekeeping gene (PBGD) and nitric oxide synthase II (NOSII) mRNA in arterial endothelial cells (AEC), bronchiolar epithelial cells (BEC), alveolar septum containing monocytes/macrophages (AS+), alveolar septum without monocytes/macrophages (AS–) and intraluminar alveolar macrophages (AM) could be quantified by real-time RT-PCR. The strongest upregulation of NOSII mRNA occurred in AM, while minimal NOSII expression was detected in BEC, AS+ and AS–. In AEC NOSII mRNA was not detectable. Conclusion: The combination of laser microdissection and real-time RT-PCR is a valuable tool for the quantitative in situ characterization of differential gene expression within complex tissues.
The Effect of Renin-Angiotensin Axis Inhibition on Early Atherogenesis in LDL-Receptor-Deficient Mice
Authors: Sharabi Yehonatan , Grossman Ehud , Sherer Yaniv , Shaish Aviv , Levkovitz Hana , Bitzur Rafael , Harats Dror
Published: April 2003

Objective: The renin-angiotensin system may play a role in the development of atherosclerosis. Nevertheless, different results from studies attempting to attenuate the process by inhibiting the converting enzyme were equivocal, and in those who succeeded, blood pressure was lowered and/or the lipid profile was improved in addition to the inhibition of the renin-angiotensin axis. The aim of this study is to investigate the effect of low doses of fosinopril, a converting enzyme inhibitor, on the development of atherosclerosis in LDL-receptor-deficient mice. Methods: Three groups of 15 mice were fed a high-fat, high-cholesterol western diet. The three study groups received either distilled water (control group), or water supplemented with fosinopril 0.01 mg/kg/day (low-dose group) or with 0.1 mg/kg/day (high-dose group). Plasma aldosterone levels and lipid profiles were measured at the beginning and at the end of the study. After 10 weeks, the mice were sacrificed and the extent of atherosclerosis was assessed at the aortic sinus. Results: Plasma aldosterone levels did not change in the control group, but decreased significantly in both treated groups from 74.7 to 39.3 ng/ml in the low-dose group (p < 0.003) and from 70.7 to 33.6 ng/ml in the high-dose group (p < 0.001). The lipid profile at the end of the study showed significantly lower levels of cholesterol and triglycerides in the high-dose group as compared to the low-dose group (p < 0.05). There was no difference between the three groups regarding the area of atherosclerosis at the aortic sinus: 157,000 ± 34,000, 130,000 ± 58,000 and 145,000 ± 26,000 µm2 in the control, low-dose and high-dose groups, respectively. Conclusion: Inhibition of the renin-angiotensin-aldosterone axis by itself does not prevent the development of early atherosclerosis in LDL-receptor-deficient mice.
Ontogeny of Pig Discrete Peyer’s Patches: Distribution and Morphometric Analysis
Authors: Makala L.H.C. , Kamada T. , Nishikawa Y. , Nagasawa H. , Igarashi I. , Fujisaki K. , Suzuki N. , Mikami T. , Haverson K. , Bailey M. , Stokes C.R. , Bland P.W.
Published: April 2003

We investigated the development of lymphoid and non-lymphoid cells in discrete Peyer’s patches (PP) of the pig using immuno-histology and image analysis. In newborn piglets discrete PP were mainly populated by CD2+, CD3+ T cells, and major histocompatibility complex class II+ cells, many of which were of macrophage and dendritic cell lineage. Four days after birth, cells were localized in defined regions: the follicle; the inter-follicular area and the dome region. Compartmentalization within the follicle started about 6 days after birth. The first signs of secondary follicles were seen from about 14 days. The pig discrete PP attained their mature structure at about 3 weeks after birth. Here we show that despite the demonstration at birth of the cell types that support antigen processing and presentation, PP did not fully differentiate morphologically until at least this time when antigen can be handled in an efficient manner.
Chronologic Changes of Cerebral Ventricular Size in a Transgenic Model of Hydrocephalus
Authors: Hayashi Nakamasa , Leifer David W. , Cohen Alan R.
Published: April 2003

We have maintained a transgenic mouse model of hydrocephalus created to overproduce the cytokine, transforming growth factor-β1 (TGF-β1) in the central nervous system (CNS). The aim of the present study was to estimate the embryonic period when the transgenic mice would develop hydrocephalus, by investigating the chronological developmental changes of the cerebral ventricles. Qualitative analysis of ventricular size was performed on sections cut in the coronal plane of embryos at the 15th (E15) and 18th (E18) embryonic days, and postnatal mice aged 4 days (P4). The presence of the TGF-β1 transgene was determined by performing polymerase chain reaction (PCR) analysis. We have examined 24 embryos and 14 postnatal mice. By performing PCR analysis, the TGF-β1 transgene was determined to be present in 16 (42.1%). Five of 16 embryos at E15 carried the transgene, and showed a slight enlargement of the lateral ventricles. Three of 8 embryos at E18 carried the transgene, and had remarkable enlargement of the lateral ventricles. Eight of 14 pups at P4 carried the transgene, and 7 of 8 pups with the transgene developed hydrocephalus. In pups that were positive for the transgene, massive enlargement of the lateral ventricles was observed and there was an associated thinning of the overlying cerebral cortex. These results suggest that congenital hydrocephalus may develop at an important embryonic time period, which coincides with the stage of neural stem cell proliferation and differentiation in the CNS.