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Biomedical subjects

R Romeo

Publications and source records attributed to R Romeo.

At least 19 recordsLinked to original sources

Surgical treatment of nasal septal perforations. Our experience.

Septal perforations are difficult problems that the otolaryngologist has to solve. In fact, the otolaryngologist has to identify the cause, which in most cases is either iatrogenic or idiopathic, to decide upon the need for surgery, and select the most suitable surgical technique of those currently available, for the case under consideration. All surgical procedures, aimed at repair of nasal septal perforations, are based on two main principles, namely repair using mucosal, mucoperichondrial, and/or mucoperiosteal flaps from the nasal cavity, or with connective tissue autograft, to be interposed between the mucosal flaps. Surgical repair of septal perforation can be carried out using either the "closed technique" or "open technique". The advantage of the former is that it does not leave any external scar. However, drawbacks related to difficulties due to the narrow operating field may be encountered. Many surgeons prefer the "open" technique, as it offers a wider operating field, thus allowing better access to the superior and posterior margins of the perforation (especially in large and/or posterior perforations), and offering binocular vision. The present report focuses on a short critical examination of the various surgical procedures described in the literature, in the attempt to identify, based also on personal experience, the most suitable techniques to repair septal perforations. A novel technique is presented for the surgical treatment of some types of perforations, which has not, so far, been described in the literature.

Adult↗

Assessment of Cottle's areas through the application of a mathematical model deriving from acoustic rhinometry and rhinomanometric data.

OBJECTIVES: Each nasal area, as defined by Cottle, has a different influence on the nasal airflow. The longitudinal distribution of resistances in nasal cavities was calculated by the anterior rhinomanometry and acoustic rhinometry data. DESIGN: Dynamic study of Cottle's areas in normal subjects was carried out by rhinomanometry and acoustic rhinometry. SETTING: Study by the Department of Otolaryngology of the University of Rome-La Sapienza. PARTICIPANTS: Twenty-seven Caucasian adults in local and general healthy conditions took part and completed this study, with a total of 54 nasal cavities included because of negativity at ENT-examination and clinical history, with normal respiratory parameters at the rhinomanometry and acoustic rhinometry. MAIN OUTCOME MEASURES: We determined nasal and acoustic resistances, nasal volumes and cross-sectional surface areas, as defined by Cottle, using nasal endoscopy. The longitudinal distribution of nasal resistances was obtained by integrating experimental surface areas using a novel mathematical model. The estimation of the longitudinal nasal resistance variations as a result of a theoretical reduction of the surface areas. RESULTS: The reduction of the 2-3-1 areas (in this order of importance) showed the greatest influence on the nasal resistances with coefficients of determinations greater than 0.98, this being quite different from that of the areas 4 and 5 for quite smaller area reduction percentages. CONCLUSIONS: The areas 2-3-1 control the overall nasal resistance so the surgical procedures on these areas greatly influence the dynamics of nasal airflow. The mathematical model developed here gives useful information to nasal functional surgery and may be applied to other schemes of nasal cavity.

Acoustics↗

Burkitt-like lymphoma of the sphenoid sinus: case report.

Burkitt's lymphoma is a malignant endemic neoplasia with a mandibular localization, described for the first time in 1958, in African children. The World Health Organization classification recognises several variants of Burkitt's lymphoma; all are highly malignant B cell lymphomas. Besides Burkitt's sporadic, endemic lymphoma and Burkitt's lymphoma associated with AIDS, the World Health Organization classification includes an "atypical or pleomorphic" variant of Burkitt's lymphoma. This subtype includes those cases diagnosed as "Burkitt-like" lymphoma in the REAL (Revised European-American Classification of Lymphoid Neoplasm). The therapeutic protocol is similar to that used for classic Burkitt's lymphoma, with chemotherapy being standard treatment. Prognosis is extremely poor, with a mean survival of < 1 year. The case is described of a sinus-nasal "Burkitt-like lymphoma", originating within sphenoid sinus. The extremely rare localisation of this histological variant of Burkitt's lymphoma is stressed as well as the extremely aggressive nature of the neoplasm.

Antineoplastic Combined Chemotherapy Protocols↗

[Paralateral-nasal malignant schwannoma with rhabdomyoblastic differentiation (Triton tumor). Report of a case].

Malignant schwannoma with rhabdomyoblastic differentiation (Triton tumor) is a very rare variant of the malignant tumors arising in the peripheral nerve sheath. In 70% of the cases, the neoplasm is associated with Von Recklinghausen's neurofibromatosis; in the remaining 30% it is the sole morbid finding. Its biological behaviour is, in any case, more aggressive than that of simple malignant schwannoma; its course is rapid and its prognosis is dismal. Local recurrences are frequent, and distant metastases, preferentially situated in the lung and brain, are present in about 50% of the cases. Post-operative radiotherapy is always indicated. We report a case of a 64-year-old man who presented a right paralatero-nasal Triton tumor. Following exeresis, a cycle of high-dose radiotherapy was prescribed; the 10-year follow-up revealed no sign of recurrence or distant metastases.

Combined Modality Therapy↗

[Horizontal glottectomy according to Calearo-Teatini for the treatment of tumors involving the anterior commissure of the larynx: oncological and functional results].

Among the different surgical options proposed for the treatment of glottic neoplasms involving the anterior commissure is the Horizontal Glottectomy (HG) proposed in 1978 by Calearo and Teatini. This type of laryngectomy is now included with good reason among the reconstructive horizontal laryngectomies in that it calls for the complete removal of the vocal folds and corresponding thyroid cartilage, with reconstruction of the airways by means of a thyrocricopexy. The object of our study was to describe the indications, surgical technique and oncological and functional results on the basis of the experience acquired by the Authors in 21 patients affected by laryngeal carcinoma (juxta-commissural T1a with diffuse hyperkeratosis of the contralateral cord and T1b infiltration to the anterior commissure) who underwent Horizontal Glottectomy (HG) according to the Calearo-Teatinin technique. For 3 of the 21 patients, the follow-up was under 3 years; for 11 patients it was over 5 years; and in 4 cases, it was over 10 years, with a survival rate at 5 years of 91.5%. There were 5 deaths (27.7%), 2 of which were due to non-neoplastic intervening causes, and 1 of which for a recurrence of N (5.5%) a year after surgery. In 3 patients, postoperative RT (radiotherapy) was carried out due to infiltration of the resection borders. With regard to the functional results, the naso-gastric tube was maintained for 8 days on average (min 4, max 12 days), and the tracheal cannula remained in situ an average of 11 days (min 7, max 19 days), while the average hospital stay was 9.3 days. The treatment options currently available for glottic tumors involving the anterior commissure may be divided among radiotherapy (RT), conventional surgery including vertical laryngectomies (Tucker's frontoanterior and Leroux-Robert's frontolateral laryngectomies) and horizontal laryngectomies (Calearo-Teatini's horizontal glottectomy, Majer-Piquet's crico-hyoido-epiglottopexy, and Labayle's crico-hyoidopexy), as well as endoscopic laser surgery. Each procedure is adopted on the basis of the experience and philosophy of the surgeon. For tumors involving the anterior commissure, our treatment protocol prescribes HG to be performed whenever possible, as its oncological and functional result have confirmed the validity of the procedure, as long as the indications are followed carefully. Furthermore, when compared to the frontolateral procedure, it provides similar oncological results, but being a much more economical operation on the vertical plane, it avoids stenotic complications, while, compared to the Majer-Piquet procedure, it allows easier and faster rehabilitation, even in elderly patients.

Aged↗

Endogenous activation of group-I metabotropic glutamate receptors is required for differentiation and survival of cerebellar Purkinje cells.

We have applied subtype-selective antagonists of metabotropic glutamate (mGlu) receptors mGlu1 or mGlu5 [7-(hydroxy-imino) cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt) or 2-methyl-6-(phenylethynyl)pyridine (MPEP)] to mixed rat cerebellar cultures containing both Purkinje and granule cells. The action of these two drugs on neuronal survival was cell specific. Although CPCCOEt (1, 10, 30 microm) reduced the survival of Purkinje cells, MPEP (3 or 30 microm) selectively reduced the survival of granule cells. Both effects required an early exposure of cultures to antagonists [from 3 to 6 d in vitro (DIV) for CPCCOEt, and from 3 to 6 or 6 to 9 DIV for MPEP]. Addition of MPEP from 6 to 9, 9 to 13, or 13 to 17 DIV also induced profound morphological changes in the dendritic tree and dendritic spines of Purkinje cells, suggesting that endogenous activation of mGlu5 receptors is required for the age-dependent refinement of Purkinje cell phenotype. In in vivo studies, an early blockade of mGlu1 receptors induced in rats by local injections of LY367385 (20 nmol/2 microl), local injections of mGlu1 antisense oligonucleotides (12 nmol/2 microl), or systemic administration of CPCCOEt (5 mg/kg, s.c.) from postnatal day (P) 3 to P9 reduced the number and dramatically altered the morphology of cerebellar Purkinje cells. In contrast, mGlu5 receptor blockade induced by local injections of antisense oligonucleotides reduced the number of granule cells but also produced substantial morphological changes in the dendritic tree of Purkinje cells. These results provide the first evidence that the development of cerebellar neurons is under the control of mGlu1 and mGlu5 receptors, i.e., the two mGlu receptor subtypes coupled to polyphosphoinositide hydrolysis.

Animals↗

Fluxional behavior of the dinitrogen ligand 2,9-dimethyl-1,10-phenanthroline in cationic methyl platinum(II) complexes.

The ionic methylplatinum(II) complexes [Pt(Me)(L)(dmphen)]X (dmphen = 2,9-dimethyl-1,10-phenanthroline, L = Me(2)SO, X = PF(6)(-) 1a, BF(4)(-) 1b, CF(3)SO(3)(-) 1c, ClO(4)(-) 1d, B(C(6)H(5))(4)(-) 1e, [B(3,5-(CF(3))(2)C(6)H(3))(4)](-) 1f; L = n-Bu(2)SO, X = CF(3)SO(3)(-) 1g; L = PPh(3), X = PF(6)(-) 2a, BF(4)(-) 2b, CF(3)SO(3)(-) 2c, ClO(4)(-) 2d, B(C(6)H(5))(4)(-) 2e, [B(3,5-(CF(3))(2)C(6)H(3))(4)](-) 2f; X = CF(3)SO(3)(-), L = CyNH(2) 3a, i-PrNH(2) 3b, 2,6-Me(2)py 3c, EtNH(2) 3d, AsPh(3) 3e, dimethylthiourea (Me(2)th) 3f and the uncharged [Pt(Me)(X)(dmphen)] (X = SCN(-) 4a, SeCN(-) 4b) complexes have been synthesized and fully characterized. In chloroform, as well as in acetone or methanol, complexes 1a-1g, 2a-2h (X = Cl(-) g, NO(2)(-) h, formed "in situ"), and 3e show dynamic behavior due to the oscillation of the symmetric chelating ligand dmphen between nonequivalent bidentate modes. All the other compounds feature a static structure in solution. The crystal structure of 2a shows a tetrahedral distortion of the square planar coordination geometry, a loss of planarity of the dmphen ligand, and, most notably, a rotation of the dmphen moiety, around the N1-N2 vector, to form a dihedral angle of 42.64(8) degrees with the mean coordination plane. The hexafluorophosphate ion lies on the side of the phenanthroline ligand. The interionic structures of 2a, 2b, and 2f were investigated in CDCl(3) at low temperature by (1)H-NOESY and (19)F[(1)H]-HOESY NMR spectroscopies. Whereas PF(6)(-) (2a) and BF(4)(-) (2b) show strong contacts with the cation [Pt(Me)(PPh(3))(dmphen)](+), being located preferentially on the side of the phenanthroline ligand, the [B(3,5-(CF(3))(2)C(6)H(3))(4)](-) (2f) ion does not form a tight ion pair. The dynamic process was studied by variable-temperature NMR spectroscopy for 1a-1f and 2a-2h in CDCl(3). The activation energies DeltaG(298) for the sulfoxide complexes 1a-1f are lower than those of the corresponding phosphine complexes 2a-2f by approximately 10 kJ mol(-)(1). The nature of the counteranion exerts a tangible influence on the fluxionality of dmphen in both series of complexes 1 and 2. The sequence of energies observed for 2a-2h encompasses an overall difference of about 16 kJ mol(-)(1), increasing in the order Cl(-) approximately NO(2)(-) << CF(3)SO(3)(-) < ClO(4)(-) < B(C(6)H(5))(4)(-) < BF(4)(-) approximately PF(6)(-) < B(3,5-(CF(3))(2)C(6)H(3))(4)(-). Acetone and methanol have an accelerating effect on the flipping. Concentration-dependent measurements, carried out in CDCl(3) for 2a with n-Bu(4)NPF(6) and the ligands dmphen, n-Bu(2)SO, sec-Bu(2)SO, and sec-Bu(2)S showed that the rate of the fluxional motion is unaffected by added n-Bu(4)NPF(6), whereas in the other cases this increases linearly with increasing ligand concentration, according to a pattern of behavior typical of substitution reactions. Dissociative and associative mechanisms can be envisaged for the observed process of flipping. Dissociation can be prevalent within the ion pair formed by a "noncoordinating" anion with the metallic cationic complex in chloroform. Among the possible associative mechanisms, promoted by polar solvents or by relatively strong nucleophiles, a consecutive displacement mechanism is preferred to intramolecular rearrangements of five-coordinate intermediates.

Journal Article↗

Structural characterization of isoxazolidinyl nucleosides by fast atom bombardment tandem mass spectrometry.

4'-Aza-2',3'-dideoxyerythrofuranosyl derivatives of thymine (AdT, 1) and uracil (AdU, 2) are analogues of 2',3'-dideoxyribofuranosyl thymine (ddT, 3) and uracyl (ddU, 4). Compounds 1 and 2 are representative of a new class of antiviral agents where the sugar moiety is replaced by an isoxazolidine ring. The increasing importance of isoxazolidinyl nucleosides has encouraged the exploitation of simple mass spectrometric rules for unambiguously assigning their structure. The species 1, 2, 5 and 6 were therefore synthesized in order to evaluate the role of the basic centre of the modified sugar moiety in their gas-phase chemistry. The tandem mass spectra of these compounds are similar to those of the wild-type nucleosides and display fragment ions corresponding to [B + 2H](+),[M - BH](+) and [B + 27](+) species, where B is the nucleobase. The last species derives from a retrocycloaddition process which is less evident in 2'-deoxyribosides. This behaviour is consistent with protonation of the analytes at the pyrimidine rings. Model isoxazolidines, in which the nucleobase was replaced by a phenyl or a naphthyl moiety, displayed the expected behaviour of species with a localized charge on the N-O moiety of the isoxazolidine ring.

Antiviral Agents↗

Targeted disruption of the nitric oxide synthase 2 gene protects against ischaemia/reperfusion injury to skeletal muscle.

To provide definitive insight into the complicated roles of the nitric oxide synthase (NOS) enzymes in ischaemia/reperfusion (I/R) injury of skeletal muscle, experiments were undertaken in mice with targeted disruption of the inducible NOS (NOS-2 KO) isoform, compared with the wild-type mouse strain. The degree of I/R injury in the NOS-2 KO mice was attenuated relative to that in the wild-type strain. After 70 min of ischaemia (24 h reperfusion), nitroblue tetrazolium (NBT) staining of skeletal muscle showed significant necrosis (40%) in wild-type mice, whilst in NOS-2 KO mice, ischaemia could be prolonged to 90 min before significant necrosis (38%) was apparent. Specific enzyme activities of the mitochondrial respiratory chain enzymes, measured in skeletal muscle homogenates, suggested that direct inhibition of the enzymes is not causal in the I/R injury. Immunohistological examination of skeletal muscle for NOS-2 showed its induction selectively in mast cells. In vitro experiments using bone marrow-derived mast cells showed that NOS-2 induction was associated with increased degranulation of mast cells. These findings suggest that NO generated by induction of NOS-2 has a deleterious effect in I/R injury of skeletal muscle and that NO exerts its damaging effect through factors released by degranulation of mast cells.

Animals↗

Comparative immunohistochemical analysis of the expression of cytokeratins, vimentin and alpha-smooth muscle actin in human foetal mesonephros and metanephros.

The human mesonephros is currently regarded as a simplified version of the foetal metanephros, primarily due to the close morphological resemblance between these two structures. The aim of the present study was to define whether human mesonephric and foetal metanephric nephrons share immunophenotypical traits in their corresponding structures (glomeruli, proximal and distal tubules). For this purpose we first investigated immunohistochemically the overall expression and topographical distribution of cytokeratins 7, 8, 18, 19, and 20, vimentin and alpha-smooth muscle actin in mature mesonephric nephrons and compared the results with those obtained in maturing-stage foetal metanephric nephrons. No expression of cytokeratins 7 and 20 was found. Cytokeratins 8, 18, and 19 and vimentin showed a restricted and basically coincident expression along the different components of both mesonephric and metanephric nephrons. These findings indicate that the intermediate filament protein profile of human mature mesonephric nephrons closely recapitulates that observed in developing metanephros and thereby strengthens the concept that human mesonephros, a transient ontogenic structure, is largely similar to the foetal metanephros. The sole difference between human mesonephros and foetal metanephros was the divergent expression of alpha-smooth muscle actin. This protein exhibited an increasingly accentuated mesangial expression paralleling the morphological maturation of metanephric glomerulus, whereas it was absent from the mesonephric one. This would suggest that the mesangial cells in these two renal structures have a different function during the foetal life.

Actins↗

Study and application of a mathematical model for the provisional assessment of areas and nasal resistance, obtained using acoustic rhinometry and active anterior rhinomanometry.

Nasal resistance (NR) depends on the geometrical features and tortuosity of the nasal airway and on the air flow. Knowing the longitudinal distribution of cross-sectional areas (CSAs) in the nasal cavity (which can be obtained using acoustic rhinometry) and the laminar nasal resistance (obtainable by processing the rhinomanometric results), it is possible to calculate, utilizing a mathematical model elaborated on the basis of fluid dynamics, the differential nasal resistance (NRdiff) and the cumulative nasal resistance (NRcum), thus localizing the position at which the highest resistance is concentrated and the related longitudinal distribution. Using a mathematical model, we integrated the sigmoid curves DeltaP/Q of rhinomanometry with the cross-sectional areas obtained using acoustic rhinometry, thus obtaining the normal distribution of differential and cumulative nasal resistances. Afterwards, we empirically reduced the cross-sectional areas corresponding to the head, body, tail and the whole inferior turbinate, recalculating the differential and cumulative nasal resistance distribution curves. The results show that reduction of up to 50% of cross-sectional areas does not substantially affect the resistivity role of the nasal valve, while greater reductions move the highest resistivity point to an area at the junction of the body and the head of the inferior turbinate. The study of the differential nasal resistance trend curves as a function of the reduction of cross-sectional areas shows that the resistance variation of the body and the whole inferior turbinate prevail with reductions of up to 40%, while the variation of cross-sectional areas of the body bordering the inferior turbinate head is predominant with higher reductions. The cross-sectional areas of the nasal airway cavity with highest resistivity are mainly located in an anterior position, where the differential nasal resistances are higher, but there are substantial variations produced by reducing the cross-sectional area of the posterior nasal airway. A similar model can produce provisional values for the results obtainable with functional nasal surgery.

Acoustics↗

Expression of leukemia inhibitory factor in human nerve following injury.

In animal models of peripheral nerve injury, leukemia inhibitory factor (LIF) is normally expressed at very low levels. Following nerve injury, its expression is rapidly increased in the nerve at the injury site and promotes both sensory and motor neuron survival. Once normal nerve function is restored, LIF expression returns to negligible levels. For this reason, LIF is considered to be a peripheral nerve trauma factor. We wished to determine whether LIF is also upregulated in human nerves following trauma and whether it is expressed in neuromas of varying age. Immunohistochemical staining for the presence of LIF was performed on injured and control human nerves from a number of subjects. Results demonstrate that LIF expression is increased in nerves within hours of injury and, in the case of neuroma formation, can persist for several years. LIF immunoreactivity was consistently found in Schwann cells, in peripheral nerve axons, and, at stages when an inflammatory response was present, also in neutrophils, mast cells, macrophages, and blood vessel walls. The level of staining within the connective tissue of injured nerves was elevated compared to control nerves, which may be due to the presence of LIF bound to the soluble secreted form of the LIF receptor. Whether the continued expression of LIF is unhealed injured nerves promotes the development of neuromas remains to be resolved.

Axons↗

Stimulating effect of an arteriovenous shunt on the in vivo growth of isografted fibroblasts: a preliminary report.

Isogenous fibroblasts derived from the skin of inbred Sprague-Dawley rats were cultured in vitro, labeled with bisbenzamide (BB) or carboxyfluorescein diacetate (CFDA), and seeded into polycarbonate growth chambers. After 24 h incubation in vitro, the chambers, either empty or containing an arteriovenous (AV) shunt, were implanted subcutaneously into the inguinal region of Sprague-Dawley rats and examined by fluorescence microscopy 2 or 7 days later. The AV shunt remained patent in all experiments. The density of labeled cells on the chamber surface in all chambers decreased in the first 2 days after insertion. At 7 days, the cell density in the empty chambers had not altered from the 2-day level, but the density in the AV shunt containing chambers had increased to almost three times the day 2 level (p = 0.013). It appears that an AV shunt can induce a significant proliferation of fibroblasts implanted adjacent to it. For at least 7 days after labeling, BB and CFDA provide a simple and effective method of quantitative detection of implanted fibroblasts. It is concluded that nutrients from the AV shunt implanted in a growth chamber result in a significant increase in the number of viable, matrix-synthesizing cells, compared with AV shunt-free controls.

Animals↗

Aneuploidy rate in spermatozoa of selected men with abnormal semen parameters.

A large proportion of patients with oligoasthenoteratozoospermia (OAT) have an abnormal karyotype and hence they produce aneuploid gametes. However, a normal karyotype does not exclude the chance of having germ cell aneuploidy, since an altered intra-testicular environment not only damages spermatogenesis, but may also disrupt the mechanisms controlling chromosomal segregation during meiosis. Therefore, this study was undertaken to evaluate the rate of aneuploidy in the spermatozoa of selected patients with abnormal sperm parameters. For this purpose, sperm aneuploidy rate for chromosomes 8, 12, 18, X and Y was evaluated by multicolour fluorescence in-situ hybridization (FISH) in nine patients with teratozoospermia alone and 19 OAT patients of presumably testicular origin. Thirteen normozoospermic healthy men served as controls. Patients with teratozoospermia or OAT had significantly greater disomy and diploidy rates compared with controls, whereas the rate of nullisomy was similar. XY disomy was very low in all groups, suggesting that chromosomal non-disjunction occurs mainly during the second meiotic division. Autosome 12 disomy rate was low in both patients and controls. There was a marked variability of total sperm aneuploidy rate in both groups of patients. Sperm aneuploidy rate was negatively correlated with sperm concentration and particularly with the percentage of normal forms. In conclusion, patients with teratozoospermia or OAT have an increased rate of sperm aneuploidy. This increase is similar in both groups, suggesting that teratozoospermia may be the critical sperm parameter associated with aneuploidy.

Adult↗

Spontaneous regression over time of the germinal epithelium in a Y chromosome-microdeleted patient: Case report.

Azoospermia factor (AZF) region microdeletions, which account for about 10-15% of patients with oligoazoospermia, seem to lack a close genotype-testicular phenotype correlation. Although many genetic and non-genetic factors may contribute to this outcome, it was thought that a spontaneous regression of testicular germ cells might also play a relevant role. The opportunity for carrying out two different testicular biopsies one year apart in an AZFc-microdeleted patient enabled corroboration of this possibility. Indeed, the first biopsy showed a spermatocyte maturation arrest with mean Johnsen scores of 4 and 3.9 in the right and left testes respectively. One year later, the right testicular biopsy showed a picture of Sertoli cell-only syndrome in 90% of the tubules examined, and of spermatogonial maturation arrest in the remaining tubules, with a mean Johnsen score of 2.1. The almost complete absence of germinal cells was confirmed by four left testicular sperm aspirations (TESA), conducted at the same time as the biopsy during an intracytoplasmic sperm injection cycle, which showed the almost exclusive presence of Sertoli cells (85% of the whole cell population). No spermatozoa could be retrieved by TESA or testicular biopsy. To our knowledge, this is the first case of a spontaneous regression of the germinal cell epithelium over time in a patient with a Yq microdeletion without the apparent intervention of any cause known to affect the germinal epithelium.

Adult↗

Asbestos exposure assessment by mineralogical analysis of bronchoalveolar lavage fluid.

Mineralogical analysis of bronchoalveolar lavage fluid (BALF) by electron microscopy can represent the most suitable method for assessing asbestos exposure. However, it has been claimed that no standardized or systematic approach to the subject of mineralogical analysis exists. This study aimed to evaluate BALF mineralogical analysis by transmission electron microscopy as biomarker of asbestos fiber load. BALF was examined in 108 exposed workers and 57 patients who underwent diagnostic fiberoptic bronchoscopy for various clinical purposes. Asbestos bodies in BALF were counted with a phase-contrast microscope. Fibers were counted and analyzed by transmission electron microscopy, which showed a significant difference between the two populations and positive results for all exposed subjects. Only 82.2% of the exposed population tested positive for asbestos bodies. Subjects with long-term exposure had higher concentrations of fibers than did those with more recent exposure, probably because of the higher workplace exposure levels in the past. The results of the study confirm that fiber concentration in BALF can be considered as a reliable biomarker of past asbestos exposure, even many years after the end of exposure.

Adult↗

The influence of extracellular matrix on the generation of vascularized, engineered, transplantable tissue.

In a recently described model for tissue engineering, an arteriovenous loop comprising the femoral artery and vein with interposed vein graft is fabricated in the groin of an adult male rat, placed inside a polycarbonate chamber, and incubated subcutaneously. New vascularized granulation tissue will generate on this loop for up to 12 weeks. In the study described in this paper three different extracellular matrices were investigated for their ability to accelerate the amount of tissue generated compared with a no-matrix control. Poly-D,L-lactic-co-glycolic acid (PLGA) produced the maximal weight of new tissue and vascularization and this peaked at two weeks, but regressed by four weeks. Matrigel was next best. It peaked at four weeks but by eight weeks it also had regressed. Fibrin (20 and 80 mg/ml), by contrast, did not integrate with the generating vascularized tissue and produced less weight and volume of tissue than controls without matrix. The limiting factors to growth appear to be the chamber size and the capacity of the neotissue to integrate with the matrix. Once the sides of the chamber are reached or tissue fails to integrate, encapsulation and regression follow. The intrinsic position of the blood supply within the neotissue has many advantages for tissue and organ engineering, such as ability to seed the construct with stem cells and microsurgically transfer new tissue to another site within the individual. In conclusion, this study has found that PLGA and Matrigel are the best matrices for the rapid growth of new vascularized tissue suitable for replantation or transplantation.

Animals↗

Expression of metabotropic glutamate receptors in the rat and human testis.

The G protein-coupled receptor kinase type 4 mediates the homologous desensitisation of type-1 metabotropic glutamate (mGlu1) receptors and is predominantly expressed in the testis. Hence, we searched for the expression of mGlu1 or other mGlu receptor subtypes in rat and human testes. RT-PCR analysis showed the presence of mGlu1, -4 and -5 (but not -2 or -3) receptor mRNA in the rat testis. The presence of mGlu1 and -5 (but not mGlu2/3) receptor proteins was also demonstrated by Western blot analysis. In the rat testis, both mGlu1a and -5 receptors were highly expressed in cells of the germinal line. It is likely that these receptors are functional, because the agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid, was able to stimulate inositol phospholipid hydrolysis in slices prepared from rat testes. Immunocytochemical analysis of bioptic samples from human testes showed a high expression of mGlu5 receptors inside the seminiferous tubuli, whereas mGlu1a immunoreactivity was restricted to intertubular spaces. mGlu5 receptors were also present in mature spermatozoa, where they were localised in the mid-piece and tail. This localisation coincided with that of beta-arrestin, a protein that is critically involved in the homologous desensitisation and internalisation of G protein-coupled receptors. Taken collectively, these results offer the first evidence for the expression of any glutamate receptor in testes, and suggest that at least mGlu5 receptors are present and functionally active in mature human sperm.

Animals↗