bluuebunny/crossref_metadata_2025 huggingface dataset:
Semen Analysis in the Usher Syndrome Type 2A
Authors: van Aarem A. , Wagenaar M. , Tonnaer E. , Pieke Dahl S. , Bisseling J. , Janssen H. , Bastiaans B. , Kimberling W. , Cremers C.
Published: April 2003

Semen analysis in patients with Usher syndrome suggested that defective connecting cilia axonemes may be involved in the irreversible, progressive loss of photoreceptors in Usher’s syndrome. In the framework of clinical genetic research into Usher syndrome, a pilot study was set up to test these findings. The semen of 6 Usher 2A patients was analysed. The fertility status of the study group of Usher 2A patients was evaluated, including semen analysis, supplemented by electron microscopic examination of the spermatozoa. Except for a significantly increased pH value, no abnormalities were found in the functional semen analysis, whereas electron microscopy revealed microtubular tail abnormalities. The latter finding was of little relevance, however, in view of the normal motility of the spermatozoa observed in these patients. There were no fertility problems in our group of Usher 2A patients, nor have any been mentioned in Usher patients in general. Earlier study findings were not supported by our data.
Aspirin Intolerance in Patients with Chronic Sinusitis
Authors: Gosepath Jan , Hoffmann Frank , Schäfer Dirk , Amedee Ronald G. , Mann Wolf J.
Published: April 2003

Aspirin intolerance in patients with chronic sinusitis is often a cause of early recurrence of symptoms after surgical treatment. This study assesses 84 patients who were tested for acetylsalicylic acid intolerance after presenting with symptoms like chronic rhinosinusitis, sometimes bronchial asthma, coexisting allergies or a history of aspirin sensitivity. Nasal polyposis was found in a majority of cases, often recurrent after previous surgery. The levels of eicosanoids such as peptido-leukotrienes and prostaglandin E2 were analyzed in isolated blood cells and compared with a healthy control group. Aspirin-intolerant patients showed elevated basal levels of peptido-leukotrienes and reduced basal levels of prostaglandin E2. Test results were graded in a system ranging from positive (68%), signifying aspirin intolerance, to borderline (18%) and negative results (14%). After screening patients with clinical findings indicating a possible aspirin intolerance, the results of this investigation reveal a strong correlation between the clinical symptomatology and the in vitro parameters of eicosanoid levels in isolated blood cells, indicating the need to induce aspirin tolerance to reduce the risk of recurrent rhinosinusitis.
Diagnosis and Physiopathology of Laryngeal Deposits in Autoimmune Disease
Authors: Hosako-Naito Yuki , Tayama Niro , Niimi Seiji , Aotsuka Shinichi , Miyaji Mamiko , Oka Teruaki , Fujinami Matuyo , Kitahara Nobuo
Published: April 2003

We report the clinical features and pathology of a previously unreported form of vocal fold disease seen in 4 patients, 3 of whom were diagnosed as autoimmune disease. The characteristic features of these lesions were found as bilateral transverse lesions in the mid portion of the vocal folds. The patients had dysphonia and diplophonia. Stroboscopic examinations showed 180-degree phase shifts between the anterior and posterior portion of the vocal folds. Case 1 had systemic lupus erythematosus, case 2 had Hashimoto’s thyroiditis, and case 3 had progressive systemic sclerosis. Prior to the onset of hoarseness, autoimmune antibody titers were increased. These cases need differential diagnosis from vocal fold nodules and cysts. Two cases were recurrent after endoscopic surgery, 1 recurring 3 times. Glucocorticoid was effective in preventing the recurrence in the early phase. Recurrence may have occurred because the surgical strategy was the same as that used for vocal fold nodules or cysts.
Intraoperative Mitomycin C in Endolymphatic Sac Surgery for Ménière’s Disease: A Pilot Study
Authors: Yazawa Yoshiro , Suzuki Mikio , Kitano Hiroya , Kitajima Kazutomo
Published: April 2003

The long-term results of endolymphatic sac (ES) shunt surgery in Ménière’s disease have been reported to be unsatisfactory compared to the short-term results, probably because of fibrosis and/or reclosure of the incised ES. To solve this problem, we tried to apply mitomycin C (MMC) intraoperatively to the incised ES as used in trabeculectomy for glaucoma. MMC has an antiproliferative effect as well as an antineoplastic effect. Thus, it is expected that MMC would have a benefit to prevent rapid fibrosis and/or reclosure of the ES. MMC did not show any ototoxicity in our animal study and we started a clinical trial after being approved by the Ethics Committee for Human Research at our university. Fourteen patients with Ménière’s disease underwent ES mastoid shunt surgery with intraoperative application of MMC and they were followed up more than 6 months. No patients experienced vertigo, although some complained of slight dizziness. Five patients out of 14 showed remarkable hearing improvement by more than 10 dB, accompanied with decrease of tinnitus.
A Possible Case of Saccular Endolymphatic Hydrops
Authors: Seo Toru , Yoshida Keiko , Shibano Akiko , Sakagami Masafumi
Published: April 2003

The vestibular-evoked myogenic potential (VEMP) is expected to be one of the examinations for saccular function. We report a case of dysequilibrium possibly due to a saccular disorder. A 27-year-old female came to our clinic complaining of severe recurrent dysequilibrium spells for few hours on September 21, 1998. The first attack was only vertigo with bilateral tinnitus for 8 days before. She did not show possible vestibular findings on a routine vestibular test. An electrochochleogram showed the dominant negative summating potential (SP) on both sides, and the furosemide test showed negative findings on both sides. A VEMP test indicated a low amplitude in the right ear, which improved to the normal response after furosemide injection. The amplitude was increased by furosemide administration even in the left ear. These findings suggested bilateral endolymphatic hydrops in the cochlea and saccule especially on the right side. The VEMP test may reveal saccular hydrops, which differs from the conventional Ménière type vertigo.
Nonlinear Mechanics of the Inner Ear and Its Relation to Otoacoustic Emissions: Two Steps on the Way to a Mathematical Model of DPOAE Generation
Authors: Ganz Michael , Berg Michael F.
Published: April 2003

Among clinical users of the registration of distortion product otoacoustic emissions (DPOAE), the understanding of the basic causality and interpretation of the phenomenon is not yet widely spread, nor is the expected influence of the middle ear and ear canal clear. On the other side, the effort in mathematical modeling of middle and inner ear structures is driven very far by now. We are convinced, though, that the essentials of an effect as DPOAE generation must be understandable from quite simple models. In a first step de Boer’s one-dimensional model was adopted and expanded by a weak frictional and a weak elastic nonlinearity, respectively. By means of perturbation theory the weakly nonlinear problem is converted in an approximation series of linear problems. So it is solvable by the common methods of linear differential equations (DEs), above all the superposition principle can be used. At the same time a structure of causality is introduced: Sources for outgoing waves are in first order approximation formed by incoming waves, and so they can be localized. The calculations show clearly that of all six cubic distortions only the 2f1 – f2 term does have a source in its ‘allowed’ region and so can travel outward. We can use the calculated DPOAE to study the influence of middle ear, external ear canal and probe plug. Some problems remain: the weakly nonlinear model in first order does not give account for proper Ldp = f(L1, L2) and Ldp = f(f2/f1) dependency, nor does it deliver additional sources or the effect of additional suppressor tones. In a second step, therefore, we replace de Boer’s simple model basilar membrane (BM) by a doubly resonant, coupled tectorial/basilar membrane (TM/BM) system. By feedback now we introduce a strong nonlinearity, which we can mathematically care for by an iterative feedback loop. The algorithm shapes the incoming waves according to strong compressive nonlinearity. More relastic incoming waves yield better source terms, and after optimization of the mistuning function between TM and BM the model now is able to deliver qualitatively correct Ldp (L1,L2) and Ldp(f2/f1) dependencies.
A Physiological Ear Model for the Emulation of Masking
Authors: Baumgarte Frank
Published: April 2003

Auditory masking is exploited in technical audio systems for transmission and storage to improve the perceived audio quality. In this respect, perception models are necessary for a masked threshold prediction to quantify the threshold of audibility for a given signal or distortion. The presented ear model emulates those parts of the auditory sound processing which are relevant to masking. The physiological modeling approach is not limited to simple masker-test-signal configurations as usually employed in psychoacoustical measurements, but it is valid for complex signals as well, which is desired in the aforementioned applications. The ear model includes a nonlinear active cochlear model which emulates the basic mechanisms effective in masking: excitation and suppression. Nonlinear mechanical amplification by means of the outer hair cells plays a crucial role in proper modeling of these effects. In addition to physiological measurements at the mechanical level of sound processing in the cochlea, classical masking patterns and nonlinear masking effects are demonstrated by the ear model.
Ultrastructural Evidence for Protection of the Outer Hair Cells of the Inner Ear during Intense Noise Exposure by Application of the Organic Calcium Channel Blocker Diltiazem
Authors: Heinrich Ulf-Rüdiger , Maurer Jan , Mann Wolf
Published: April 2003

Outer hair cells could be protected during intense noise exposure by the application of the calcium channel blocker diltiazem given before and after noise treatment. After various experimental approaches, the ultrastructural morphology was analysed for the different animal populations in the basal part of the second turn of the cochlea, which was the most destroyed area after an acute noise trauma caused by a gun shot (sound pressure at the ear drum 156 dB, frequency maximum between 4 and 6 kHz). Compared to untreated control specimens (experimental animal group I), the outer hair cells in the basal part of the second turn of the cochlea were mostly destroyed without any diltiazem application but after intense noise exposure (group II) or when the calcium channel blocker was given only before (group III) or only after an intense noise exposure (group IV). Only cellular debris of these receptor cells could be identified within this region of the cochlea at the electron-microscopic level. After diltiazem application before and after the intensive noise exposure (group V), the general morphology of the tissue was much better conserved and almost no destroyed outer hair cells were found. But nevertheless, the electron-microscopic analysis revealed that the cellular fine structure was altered. Compared to untreated control specimens, small invaginations were sometimes seen in the basolateral membranes of outer hair cells, and also the cuticular structures of the Deiters cells forming the lamina reticularis bulged into the outer hair cells. To get more information about possible physiological alterations, the amounts of precipitable calcium within the outer hair cells were determined by an image-processing system for the five different populations (groups I–V). The values obtained were compared to the calcium content in specimens which had received diltiazem without any additional noise exposure (group VI) and with an animal population which was exposed to a pure tone for 5 min at 90 dB (group VII). The results are discussed in respect of outer hair cell protection preventing morphological damage and/or physiological alterations.
Real-Ear to Coupler Difference in Patients with Ear Drum Perforation
Authors: Liu Tien-Chen , Lin Kai-Nan
Published: April 2003

The aim of this study is to investigate the effects of ear drum perforation on real-ear to coupler difference (RECD) in adults. RECD was measured using a probe tube microphone system in 22 patients with ear drum perforations. Twenty-two normal subjects served as controls. For normal subjects, RECD was in good agreement with the values reported in the literature. For the perforated ears, the RECD was up to 8 dB smaller in the frequency range from 0.5 to 1 kHz. There was no significant difference at frequencies below 0.25 kHz and above 1 kHz. A much larger intersubject variability was found in the experimental group. The mean intersubject standard deviation was 4.4 dB in the experimental group as contrasted with 2.2 dB in the control group. Neither the equivalent ear canal volume nor the perforation size appeared to be correlated with the degree of RECD reduction over lower frequencies. These results strongly suggest the need for individual RECD measurements, rather than using the average normal RECD, to appropriately compensate for the reduced transmission of lower-frequency sounds in fitting hearing aids for patients with ear drum perforations.
Relationship between Neurological Asymptomatic Vertigo and the Vertebrobasilar System as Revealed by Magnetic Resonance Angiography
Authors: Seo Toru , Tominaga Satoru , Sakagami Masafumi
Published: April 2003

Many patients suffering from vertigo have no neurological symptoms except for a positional nystagmus. Vertigo without any neurological findings has not been thought to be a vertebrobasilar (VB) attack. The purpose of this study is to clarify the relationship between vertigo without any neurological symptoms and the VB system using magnetic resonance angiography (MRA). MRAs of the VB system were examined in 31 patients with positional nystagmus of unknown origin (PNU) and in 14 patients with benign paroxysmal positional vertigo (BPPV) as control. MRA was performed with a 1.5-tesla system using the two-dimensional time-of-flight technique. Abnormalities such as elongation, bending, narrowing or obstruction of the artery were detected in 27 patients of the PNU group and 8 patients of the BPPV group. A significant difference in the abnormalities was noticed between the two groups (p < 0.05; χ2 test). This result is almost similar to those of previous studies using conventional vertebral angiography and digital subtraction angiography. Thus, MRA is useful in examination of the VB system. Moreover, our study suggested that the positional nystagmus might result from VB ischemia, even if there were no other neurological signs.
The Effects of Chronic Electrical Stimulation on Laryngeal Muscle Physiology and Histochemistry
Authors: Zealear David L. , Billante Cheryl R. , Chongkolwatana Cheerasook , Rho Young S. , Hamdan Abdul-Latif , Herzon Garrett D.
Published: April 2003

The present study examined the effects of functional neuromuscular stimulation (FNS) on posterior cricoarytenoid (PCA) muscle physiology and histochemistry. In 4 canines, 10 cm of the recurrent laryngeal nerve was resected. A patch electrode array was implanted for PCA stimulation. FNS was applied to 2 canines for a period of 4 weeks with 2 additional animals serving as nonstimulated controls. Results indicated that FNS increased PCA muscle contractility over the period of intervention but had no effect on contraction speed. FNS also protected the muscle from atrophy by preventing muscle weight loss and type 2 fiber deterioration. Finally, it rescued muscle fibers from ensuing fibrosis.
The Effects of Chronic Electrical Stimulation on Laryngeal Muscle Reinnervation
Authors: Zealear David L. , Billante Cheryl L. , Chongkolwatana Cheerasook , Herzon Garrett D.
Published: April 2003

The present study examined the effects of functional neuromuscular stimulation (FNS) on reinnervation of the posterior cricoarytenoid (PCA) muscle. In 4 canines, the recurrent laryngeal nerve (RLN) was sectioned and anastomosed and a patch electrode array implanted for stimulation and recording at multiple PCA sites. Following implantation, FNS was applied to 2 canines for a period of 6 weeks. Two additional animals served as nonstimulated controls. In each animal, histomorphometric analysis of the RLN was used to assess the quality of nerve regeneration and the potential for muscle reconnection. The magnitude of reinnervation was monitored by electromyographic (EMG) potentials evoked by RLN stimulation. The appropriateness of reconnection was determined by the pattern of spontaneous EMG activity and recovery of vocal fold abduction. Results of this preliminary study indicated that FNS caused an overall repression of reinnervation. However, the repression preferentially inhibited reconnection by foreign nerve fibers, promoting selective reinnervation and preventing synkinesis.
Hereditary Sensorineural Hearing Loss of Unknown Cause Involving Mitochondrial DNA 1555 Mutation
Authors: Iwasaki Satoshi , Tamagawa Yuya , Ocho Shuji , Hoshino Tomoyuki , Kitamura Ken
Published: April 2003

We report on a family with maternally inherited sensorineural hearing loss, in which no history of aminoglycoside injection and no other specific etiology could be identified in any member. A 1555 A-to-G mutation of mitochondrial DNA was found in all members demonstrating hearing loss. The hearing in the propositus and his sister was severely impaired at a younger age than that in the mother. This case suggests that the 1555 point mutation of mitochondrial DNA has potential to promote inherited nonsyndromic hearing loss without any known etiology.
Intratympanic and Systemic Dexamethasone for Ménière’s Disease
Authors: Hirvonen T.P. , Peltomaa M. , Ylikoski J.
Published: April 2003

Intratympanic and systemic dexamethasone treatment of Ménière’s disease (MD) was evaluated in a prospective study. Seventeen patients (6 men and 11 women) with MD (5 right-sided, 11 left-sided and 1 bilateral) were treated with three 0.2- to 0.4-ml injections of intratympanic dexamethasone hyaluronate (16 mg/ml) during a week and with an initial intramuscular dexamethasone injection of 15 mg. Most of patients were in stage 3, and the mean duration of MD was 5.3 years. Pure-tone and speech audiometry and the symptom scale of the patients were followed up for 1 year after the treatment. Symptoms of aural fullness, hearing loss, tinnitus and vertigo did not improve significantly. However, sufficient control of vertigo was achieved in 76% of the patients. In conclusion, no definite treatment effect has yet been shown for intratympanic and systemic dexamethasone treatment. Therefore, the clinical use of dexamethasone in MD needs further investigation.
Primary Malignant Melanoma Arising from the Base of the Tongue
Authors: Misawa Hayato , Yanagita Noriyuki , Iwagaki Toshinori , Asahi Yukari , Yokoi Hisashi , Kato Kazuo , Qian Bin , Shamoto Mikihiro
Published: April 2003

A rare case of malignant melanoma of the base of the tongue in a 65-year-old Japanese woman is described. The primary and metastatic tumors were treated by surgery, chemohormone and immunotherapy. There was no recurrence for 5 years after the initial diagnosis. Electron microscopy suggested that the melanosomes of the melanoma cells in the cervical lymph nodes were less mature than those in the primary melanoma cells; melanosome immaturity was consistent with a more aggressive quality.
Mutants of p53 Gene Presence in Laryngeal Carcinoma and Adjacent Histopathologically Normal Tissue
Authors: Jin Xiaojie , Zhou Liang , Zhao Aiguang
Published: April 2003

Possible differences were analyzed between histopathological and molecular biological findings in laryngeal cancer tissue and its adjacent normal tissues. p53 gene mutations were detected in histopathologically normal tissue both within 0.5 cm of and more than 0.5 cm distant from the nearest cancerous tissue. Since the p53 mutation was common in both cancer tissue and adjacent normal tissue, the gene mutation may play an important role in laryngeal carcinogenesis. Radical excision of the tumor is defined by histopathological limits, but ‘normal’ tissue may nevertheless contain the gene mutation, giving rise to the danger of further cancer development. We suggest that the determination of the surgical margin should be based on a combination of histopathology and molecular biological findings.
Reversible Tobramycin-Induced Bilateral High-Frequency Vestibular Toxicity
Authors: Walsh Rory M. , Bath Andrew P. , Bance Manohar L.
Published: April 2003

We report an unusual case of tobramycin-induced bilateral high-frequency vestibular toxicity with subsequent clinical and objective evidence of functional recovery. In those patients with a clinical presentation suggestive of aminoglycoside-induced bilateral vestibular toxicity (ataxia and oscillopsia) and normal low-frequency (ENG-caloric) responses, high-frequency rotation chair testing should be performed to exclude a high-frequency vestibular deficit.
The New Imaging-Based Classification for Describing the Location of Lymph Nodes in the Neck with Particular Regard to Cervical Lymph Nodes in Relation to Cancer of the Larynx
Authors: Som Peter M. , Curtin Hugh D. , Mancuso Anthony A.
Published: April 2003

For over five decades, the principle landmarks used in cervical nodal classification were clinical and defined either by palpation or found at the operative table. However during the past two decades, sectional imaging has consistently improved its quality and resolution and it has been shown that imaging can identify deep structures and adenopathy not amenable to palpation. Such disease can alter planned operative or radiation fields. In the April 1999 issue of the Archives of Otolaryngology – Head Neck Surgery, for the first time an imaging-based classification was published that gave precise anatomic landmarks for use in classifying metastatic cervical adenopathy. This classification was developed in consultation with head and neck surgeons so that the nodal levels classified by this imaging-based system would correspond closely with the nodal levels determined by utilizing the most commonly employed clinically-based classifications. This article describes this imaging-based classification and demonstrates its use with axial diagrams.
The Pathology of Neck Dissection in Cancer of the Larynx
Authors: Devaney Stephanie L. , Ferlito Alfio , Rinaldo Alessandra , Devaney Kenneth O.
Published: April 2003

Cancer of the larynx is a common problem in a head and neck oncological surgical practice; as such, pathology departments supporting such surgical practices will examine cervical lymph node dissection specimens with some frequency. Issues to be settled among pathologists and surgeons include – How precise an anatomic dissection of the specimen is called for? What histological features of the specimen will be of most use to the clinicians who are devising a course of postoperative therapy for the patient? What sorts of methods are needed to identify the maximum number of micrometastases which may be lurking within the lymph nodes of the specimen? Is there a role for routine application of special techniques – such as immunohistochemistry or molecular biology – in the analysis of these specimens? While the answers to these questions are likely to vary somewhat from one center to another, patients are best served when these questions are discussed amongst the respective physicians before surgical procedures are undertaken, rather than after the fact.
Quantification of Osteoclastic Resorption of the Bovine Otic Capsule in vitro by an Enzyme-Linked Immunosorbent Assay
Authors: Frisch Thomas , Foged Niels Tækker , Sørensen Mads Sølvsten , Bretlau Poul
Published: April 2003

The bony shell surrounding the inner ear is known to have a very pronounced centripetal inhibition of remodelling in vivo, with almost no bone turnover immediately adjacent to the perilymphatic spaces and a gradually increasing turnover rate towards outer parts of the bony otic capsule. By the use of in vitro markers of bone resorption, including an enzyme-linked immunosorbent assay for quantification of type I collagen degradation and a colorimetric enzyme assay for quantification of osteoclast tartrate-resistant acid phosphatase activity, this study demonstrates that there are no ex vivo differences in bone matrix resorption between the inner and outer parts of the otic capsule when exposed to seeded osteoclasts from rabbits. Thus, the unique spatial distribution of perilabyrinthine bone turnover is not caused by a shift in resorbability from inner to outer capsular bone that is due to inherent bone quality differences particular to these bone compartments. More likely, the sustained action of some intravital ‘field force’, originating from the inner ear spaces, is responsible for the unique spatial distribution of the otic capsular bone turnover found in vivo. Though the character of this force is not yet defined, it is appealing to relate it to the large electromagnetic potential gradient present in the inner ear.
Acquired and Congenital Cholesteatoma: Determination of Tumor Necrosis Factor-alpha, Intercellular Adhesion Molecule-1, Interleukin-1-alpha and Lymphocyte Functional Antigen-1 in the Inflammatory Process
Authors: Akimoto Rika , Pawankar Ruby , Yagi Toshiaki , Baba Shunichi
Published: April 2003

The molecular and cellular factors resulting in the pathologic features of acquired and congenital cholesteatomas are not completely known. Recently, proinflammatory cytokines like interleukin-1α (IL-1α) and tumor necrosis factor-alpha (TNF-α) have been shown to induce bone resorption, in vitro. To elucidate the key molecules involved in bone resorption and cell infiltration associated with cholesteatoma, we examined the in vivo levels of IL-1α and TNF-α, intercellular adhesion molecule-1 (ICAM-1) and lymphocyte functional antigen-1 (LFA-1) in acquired and congenital cholesteatomas, by reverse transcriptase-polymerase chain reaction, immunohistochemistry, and ELISA. Increased levels of IL-1 and TNF-α were detected in both types of cholesteatomas as compared to normal skin. Increased ICAM-1 expression and LFA-1+ cells were detected in acquired but not congenital cholesteatoma. Strong correlation was detected between TNF-α and bone resorption in both types of cholesteatoma, and between TNF-α and ICAM, TNF-α and severity of infection, or cell infiltration in acquired cholesteatoma. No correlation existed between various parameters and IL-1α. These results suggest that TNF-α may play a crucial role in the pathogenesis of both acquired and congenital cholesteatomas by regulating bone resorption and cell infiltration.
Superficial Siderosis of the Central Nervous System
Authors: Takasaki Kenji , Tanaka Fujinobu , Shigeno Koichiro , Kanda Yukihiko , Kawajiri Ippei , Tashiro Tetsuya , Kobayashi Toshimitsu
Published: April 2003

This is a case of superficial siderosis of the central nervous system (SSCN). The diagnosis of SSCN was based on the result of T2-weighted magnetic resonance imaging and on suggestive clinical manifestations. The pure-tone audiogram showed bilateral progressive sensorineural hearing loss with a poor speech discrimination score and Jerger type IV. The remarkable elevation of the detective threshold of cochlear microphonics on electrocochleography was found and distortion product otoacoustic emission (DPOAE) showed no response: These electrophysiologic examinations, including electrocochleography and DPOAE, revealed that the progressive sensorineural hearing loss in this case was caused by both retrocochlear and cochlear damages.
Epithelial-Myoepithelial Carcinoma of the Nasopharynx
Authors: Imate Yuji , Yamashita Hiroshi , Endo Shiro , Okami Kenji , Kamada Tomomi , Takahashi Masahiro , Kawano Hiroo
Published: April 2003

We present a patient with a rare epithelial-myoepithelial carcinoma of the nasopharynx with a typical biphasic histopathologic pattern. Immunohistochemical findings support the concept that the inner cells differentiate to ductal epithelium and the outer cells to myoepithelium. In this patient complete excision of the tumor and reconstruction with a pedicled sternocleidomastoid myocutaneous flap were performed. A DNA diploid pattern from flow cytometric study indicates a favorable prognosis. There was no recurrence nor metastasis for 55 months.
Immunoreactivity of Adventitial Matrix Fibrils of Normal and Aneurysmal Abdominal Aorta with Antibodies against Vitronectin and Fibrinogen
Authors: Hirose Hitoshi , Ozsvath Kathleen J. , Xia Shichao , Gaetz Harold P. , Tilson M. David
Published: April 2003

Purpose: We have reported that immunoglobulin G (IgG) harvested from specimens of aneurysmal abdominal aorta (AAA) is immunoreactive with a fibrillar component of the matrix of the aortic adventitia. In further studies we have reported the partial amino acid sequence of a 40-kDa protein, purified from the adventitia of the human aorta, which we have called aortic aneurysm antigenic protein-40 kDa (AAAP-40). AAAP-40 has homologies with bovine microfibril-associated glycoprotein-36 kDa (MAGP-36). Both AAAP-40 and MAGP-36 have homologies to fibrinogen beta (FB-b) and vitronectin (VN). The purposes of the present experiments were (1) to determine whether antibodies against VN and fibrinogen are immunoreactive with elements of the normal and/or aneurysmal aortic wall, and (2) to determine whether these antibodies are immunoreactive with soluble extracts of aortic proteins. Methods: Paraffin-embedded tissue sections of normal and aneurysmal aorta were probed with polyclonal rabbit antihuman VN and antihuman FB-b antibodies. Immunoblots of soluble aortic proteins were evaluated with the same antibodies. Histochemical preparations with Gomori’s aldehyde fuchsin and elastin-von-Gieson solutions were also performed. Results: Anti-VN and FB-b antibodies reacted with matrix fibrils in the aortic adventitia in both normal and aneurysmal specimens, with a distribution that resembles the appearance of fibrils that are stained by Gomori’s reaction. By comparison to normal adventitial fibrils, fibrils in the specimens from AAA appeared fragmented, coiled, and frayed. Antibodies against VN and FB-b were immunoreactive in immunoblots with a soluble aortic protein of molecular weight ∼40 kDa, consistent with the migration of AAAP-40. Conclusions: There appear to be immunodeterminants in AAAP-40 that are recognized by polyclonal antibodies against VN and FB-b.
72-kD (MMP-2) and 92-kD (MMP-9) Type IV Collagenase Production and Activity in Different Histologic Types of Lung Cancer Cells
Authors: González-Avila Georgina , Iturría Cecilia , Vadillo Felipe , Terán Luis , Selman Moisés , Pérez-Tamayo Ruy
Published: April 2003

In this study we examined the production of gelatinases A and B (MMP-2 and MMP-9), and their natural inhibitors TIMP-1 and TIMP-2 in cell lines derived from different histologic types of lung cancer. Gelatinolytic activity was measured by zymography and radiolabeled gelatin degradation. Immunocytochemistry and Western blot analysis were performed to corroborate the presence of immunoreactive MMP-2, MMP-9, TIMP-1 and TIMP-2 proteins. The highest gelatinolytic activity was identified in the cell extracts from a small-cell carcinoma cell line. MMP-9 was observed in all samples as a proenzyme, while MMP-2 was present as zymogen in the squamous-cell and in the small-cell carcinomas, and in its active form in one squamous-cell carcinoma cell line. TIMPs were also present in the neoplastic lung cell lines. TIMP-1 was observed in the media of all cells as a 21-kD band, and as TIMP-1 polymers with the exception of the small-cell carcinoma samples. TIMP-2 was found as higher-order molecular immunoreactive complexes that may correspond to proMMP-2/TIMP-2 complexes. These results demonstrate that lung neoplastic cells produce both MMP-2 and MMP-9 and their inhibitors, with the small-cell carcinoma cell extracts showing the highest enzymatic activity. This gelatinolytic activity fits well with the clinical metastatic behavior of this type of lung cancer.
Definition of Early Progenitors and Functional Maturation of Human Natural Killer Cells: Requirements for Cytocidal Activity
Authors: Vaz Fatima , Hoffman Ronald , Almeida-Porada Graca , Ascensao Joao L.
Published: April 2003

We examined the in vitro development of human natural killer (NK) cells to define the earliest hematopoietic progenitors which could give rise to NK cells. Our data indicates that NK cells arise from the Thy 1+ subpopulations of CD34+ marrow cells. In cultures with IL2 we generated up to 96.5% NK cells from CD34+ Thy 1+ and in cultures with IL7, cells bearing NK cell surface markers were also observed but were devoid of functional activity. We also addressed the functional integrity of these cultured cells by examining the appearance of granzyme A, serine esterase, Met-ase and perforin and correlating these with lytic activity. Granzyme A mRNA could be detected in IL-2-cultured cells and serine esterase activity as well as perforin were also found at the single cell level. Human Met-ase (Hu-Met-1) was not detected at any time of analysis. Granzyme expression occurs early in cells cultured with IL2 but its presence did not correlate with lytic function; CD56+ cells grown in the presence of IL7 express these enzymes and perforin after longer periods of culture, but lack the ability to lyse NK targets.
Cellular Sequences in Stealth Viruses
Authors: Martin W. John
Published: April 2003

Cloned DNA obtained from the culture of an African green monkey simian cytomegalovirus-derived stealth virus contains multiple discrete regions of significant sequence homology (p values ranging from 4 × 10–3to 1 × 10–20) to portions of known human cellular genes. The stealth virus was cultured from a patient with chronic fatigue syndrome (CFS). Earlier studies had revealed considerable sequence heterogeneity within DNA fragments isolated from virus-infected cells. A set of polymerase chain reaction (PCR) primers generated different PCR products when tested on stealth virus cultures from 4 patients with CFS. Several of the PCR products also contain regions of significant partial homology to distinct cellular sequences, including sequences repetitively expressed throughout the cellular genome. Stealth viruses may play an important role in the origins and in the genetic diversity of both viral and cellular sequences.
Fluorescent in situ Hybridization Detection of HER-2/neu Gene Amplification in Rhabdomyosarcoma
Authors: Mark Hon Fong L. , Brown Stephen , Sun Ci-Lin , Samy Mangala , Afify Alaa
Published: April 2003

Embryonal rhabdomyosarcoma is the most common malignant soft-tissue tumor in childhood, comprising 45–50% of childhood sarcomas. Cytogenetic studies of this tumor are rare. In view of the paucity of cytogenetic data on this cancer and based on the finding of HER-2/neu gene amplification in a number of cancers that was detected mostly using the traditional technique of immunohistochemistry, we decided to conduct a pilot study to investigate whether HER-2/neu gene amplification in this tumor can be detected using the newer technique of fluorescent in situ hybridization (FISH). Archival tissues of rhabdomyosarcoma were retrieved and FISH using an HER-2/neu probe was undertaken on formalin-fixed paraffin-embedded tissue sections using a protocol optimized for our laboratory at Rhode Island Hospital. Out of 9 cases of rhabdomyosarcoma studied to date, 1 case clearly showed HER-2/neu gene amplification. Thus, FISH is a sensitive technique suitable for the detection of oncogene amplification and the delineation of tumor heterogeneity in this tumor. Future experiments utilizing additional specimens from our centers as well as from other laboratories will be needed to extend the finding in the present pilot study.
Interpreting Data on Lymphocyte Subsets in the Blood of HIV Patients – Organ Distribution, Proliferation and Migration Kinetics Are Critical Factors
Authors: Pabst R. , Rosenberg Y.J.
Published: April 2003

Lymphocytes stay in the blood only a short time before migrating to lymphoid and nonlymphoid organs. They represent only about 2% of all lymphocytes in the body. The ratio of CD4+/CD8+ lymphocytes in the blood depends on age and genetic influences. In HIV infection not only relative but also absolute numbers of lymphocyte subsets should be determined. The different effects of proliferation and apoptosis on lymphocytes in HIV infection have to be considered. Lymphocyte levels in the blood of HIV patients do not mirror alterations in the lamina propria of the gut or lymph nodes. The dynamic aspects of lymphocyte life span and migration during HIV infection and the progression to AIDS as well as the effects of treatment have to be taken into consideration to enable a meaningful interpretation of experimental data. More data from animal models of HIV infection are needed to study these kinetic aspects.
Analysis of Virologic and Immunologic Events in HIV Infection
Authors: Graziosi Cecilia , Pantaleo Giuseppe
Published: April 2003

A number of pathogenic events occurring immediately after the transmission of HIV lead to the establishment of chronic infection. In fact, despite the detection of vigorous virus-specific immune responses during primary infection, HIV is able to establish chronic infection in most of the cases. This is the result of several virologic and immunologic mechanisms that HIV has evolved to escape and/or to weaken virus-specific immune responses. Lymphoid organs represent the primary anatomic site for the establishment of chronic infection, and if highly active antiretroviral therapy is not initiated in early stage disease, there is a progressive destruction of lymphoid tissue that ultimately leads to the profound immunosuppression typical of AIDS. Therefore, analysis of lymphoid organs is crucial for the correct evaluation of the effectiveness of antiretroviral therapy in HIV infection.
Mucosal Immunodeficiency in HIV/SIV Infection
Authors: Zeitz M. , Ullrich R. , Schneider T. , Kewenig S. , Riecken E.-O.
Published: April 2003

The gastrointestinal tract is one of the major target organs for secondary infections and malignancies in HIV infection in humans indicating disturbed local immunologic defense mechanisms. Immunohistology and flow cytometric studies have demonstrated a more pronounced loss of CD4+ T cells in the intestinal mucosa compared to the peripheral blood in humans infected with HIV. In parallel, activated CD8+ T cells in the lamina propria are increased in this compartment. In SIV-infected nonhuman primates a very early loss of CD4+ T cells in the intestinal mucosa compared to the peripheral blood occurs already at 2 weeks after infection. Depletion and functional impairment of mucosal CD4+ T lymphocytes with consecutive altered cytokine secretion in HIV/SIV infection may explain the breakdown of the mucosal immune barrier leading to secondary opportunistic or nonopportunistic infections and secondary malignancies. In addition, due to the interrelation between the mucosal immune system and epithelium, these changes might be responsible for the partial small intestinal mucosal atrophy and maturational defects in enterocytes observed in HIV-infected patients.
Regulation of the Human glut4 Gene Expression in Tumor RD18 Cell Line
Authors: Chiaramonte Raffaella , Bartolini Enrico , Testolin Corrado , Comi Paola
Published: April 2003

We tested if glut4, the gene for muscle-specific glucose transporter, underwent some variations of expression in neoplastic cells. Our model was a rhabdomyosarcoma cell line (RD18) which retains the ability to differentiate along the myogenic pathway. Any definable changes of expression of glut4 in normal and RD18 cells were revealed by Northern blot analysis. In order to identify the transcriptional regulatory regions of the glut4 gene we performed a deletion analysis of the 5′ flanking region. The downregulation which we found in the expression of this gene in RD18 cells could be related with the activity of a negative regulatory element.
Tissue-Specific High-Level Expression of Human Endogenous Retrovirus-R in the Human Adrenal Cortex
Authors: Katsumata Kazuaki , Ikeda Hitoshi , Sato Masayuki , Harada Hiroshi , Wakisaka Akemi , Shibata Masahiko , Yoshiki Takashi
Published: April 2003

In an attempt to clarify the biological nature of a human endogenous retrovirus (HERV), HERV-R, which is a single-copy type of HERVs and is conserved as a full-length viral sequence, the expression of HERV-R mRNA in normal autopsied systemic organs was examined by Northern blot analysis. The expression showed different levels among individuals, with the adrenal glands expressing the highest level of HERV-R among all organs tested, except for the placenta. In various adrenal tumors, HERV-R was expressed at high levels in all cortical adenomas but less so in pheochromocytomas. In situ hybridization revealed the expression of HERV-R to be localized in all layers of the adrenal cortex, but not in the medulla. This high-level expression of HERV-R in the adrenal cortex may possibly relate to differentiation and/or steroid production by adrenocortical cells.
Prolonged Hematopoietic Chimerism in Normal Mice Transplanted in utero with Human Hematopoietic Stem Cells
Authors: Pixley John S. , Zanjani Esmail D. , Shaft Donna M. , Porada Christopher D. , Mackintosh F. Roy
Published: April 2003

We have previously reported prolonged hematopoietic chimerism in normal mice transplanted in utero with human fetal hematopoietic stem cells (HSC) by flow cytometry. We now further confirm the human origin of these cells by demonstrating human DNA in the marrow of one such chimeric mouse. We also examined 42 mice born after in utero transplantation with HSC enriched from human adult marrow cells. All live-born mice were treated with recombinant human growth factors. Twelve had human cells in the peripheral blood (range: 01.–2.93%). Thymic samples were positive in 3 cases. The bone marrow of 2 mice contained cells expressing human CD34 antigen. Light scatter characteristics support the presence of multilineage hematochimerism. Human IgM was present in 2 of 4 chimeric sera tested. Thus, normal mice transplanted in utero with human HSC may permit long-term engraftment and differentiation of the human HSC.
Expression of the MCP-1 Gene and the HPV 16 E6/E7 Oncogenes in Squamous Cell Carcinomas of the Cervix uteri and Metastases
Authors: Riethdorf Sabine , Riethdorf Lutz , Richter Nicole , Löning Thomas
Published: April 2003

Monocyte chemoattractant protein 1 (MCP-1) plays an important role in the recruitment of monocytes in solid tumors. Previously, for cervical carcinoma cell lines an inverse correlation between the expression of the MCP-1 gene and the human papillomavirus (HPV) oncogenes E6/E7 was described. In this study paraffin-embedded biopsy specimens from 25 squamous cell carcinomas of the cervix uteri were analyzed for the expression of the MCP-1 gene and the HPV 16 E6/E7 oncogenes by RNA/RNA in situ hybridization. Similar to in vitro analyses, in our material a negative correlation between the transcription of these genes in tumor cells could be demonstrated in 19/25 cases, whereas in 6 cases both MCP-1 gene and HPV oncogene transcription could be detected. Interestingly, the same results regarding MCP-1 gene and HPV oncogene expression were observed in the majority of corresponding metastases. On the other hand, in stromal cells MCP-1-specific transcripts could be detected in all cases. In most of them intra- and/or peritumoral macrophages were observed. Our findings support the hypothesis that HPV oncoproteins are negative regulators of MCP-1 transcription.
Monocyte-Dependent Stimulation of Human T Cells by Silicon Dioxide
Authors: Smalley David L. , Shanklin Douglas R. , Hall Mary F.
Published: April 2003

Mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation from randomly selected silicone breast implant recipients for testing. Restricted antibody to HLA-DR (28–33 kD) depleted the concanavalin A mitogenic response which was expected but failed to inhibit the proliferative response to silicon dioxide. Further testing with monoclonal antibodies to HLA-DP, -DQ, and a second -DR with specificity for the NS1 region of the MHC class II genome, all markedly inhibited proliferation of T cells despite otherwise adequate stimulation by concanavalin A or silicon dioxide. Monoclonal antibodies directed against B7-1 also inhibited proliferation of T cells following stimulation with concanavalin A or silicon dioxide. These results confirm the T-cell response to silicon dioxide is monocyte-dependent and not a superantigen as has been speculated.
Conventional and Molecular Cytogenetic Identification of a Variant Klinefelter Syndrome Patient with a Deleted X Chromosome
Authors: Mark Hon Fong L. , Feldman David , Sigman Mark
Published: April 2003

We report on the case of a 34-year-old patient with the Klinefelter syndrome and an unusual cytogenetic finding of a deletion involving the short arm of the X chromosome. This was confirmed with fluorescent in situ hybridization (FISH) using an X chromosome-specific whole chromosome painting probe. The patient presented with infertility. The only abnormal physical findings were atrophic testes with azoospermia and elevated levels of follicle-stimulating hormone and luteinizing hormone. This case represents a relatively mild manifestation of the Klinefelter syndrome. Previous reported cases were often associated with more severe phenotypes such as variable degrees of mental retardation and facial dysmorphism, hypothesized as due to the failure of X inactivation. The X inactivation center, located on Xq13, is presumably intact in our patient, who had a deletion involving only the short arm. The mild phenotype observed in our patient was found to be consistent with the conventional and molecular cytogenetic findings.
Synovial T Cell Receptor Heterogeneity in Early Arthritis
Authors: Fang Qiong , Sun Yan-Yang , El-Gabalawy Hani , Cai Wei , Ko Linda , Chin Howard , Arayssi Thurayya , Schumacher H.Ralph , Williams WilliamV.
Published: April 2003

Rheumatoid arthritis (RA) has been postulated to result from a synovial immune response to an unidentified antigen(s), which should be mirrored by the T cell response. Here we investigate the T cell receptor (TCR) repertoire in the synovial tissue of patients with arthritis of early to moderate duration. We developed a nested polymerase chain reaction (PCR) technique to examine the TCR repertoire of small biopsy specimens, and show that the method is highly sensitive. We apply this technique to synovial biopsies obtained from the knee joints of patients with early to moderate duration arthritis (average duration of arthritis 1 year, range 0.02–2.75 years). We examined biopsies from 5 normal individuals, 32 RA patients, 7 patients with seronegative spondyloarthropathy (Sp), and 12 patients with undifferentiated arthritis (UA). TCR message was detectable in 4/5 normals, 15/32 RA, 5/7 Sp, and 8/11 UA biopsies, with sampling error likely accounting for most negative biopsies. The average numbers of TCR Vβs detected per TCR-positive biopsy were 5.0 ± 3.7 for normals, 12.7 ± 8.4 for RA, 18.0 ± 7.4 for Sp patients, and 14.4 ± 10.2 for UA. Examination of TCR messages by single-stranded conformational polymorphism analysis showed similar proportions of dominant clones in the normals compared with the patients with inflammatory arthritis. Sequence analysis was performed on 33 dominant clones from 16 patients. Sequence alignment of the third hypervariable regions showed some evidence of disease-specific sequence clustering for Sp, while some RA sequences showed similarity to previously described motifs. These data indicate greater TCR heterogeneity in early Sp and UA compared with normal synovium. Disease-specific TCR sequences may occur in early RA and Sp.