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1.
Heliyon ; 5(12): e02932, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31867458

ABSTRACT

ODF1 has been described as an exclusively expressed testicular protein and is located in the outer dense fibers along the sperm tail. ODF1 has been involved in the sperm motility and in the development of the flagellum, but the function of ODF1 is not already clear. Other ODF proteins, such as ODF2 have been characterized in other tissues like the basal body of the kidney primary cilium, but so far only the mRNA of ODF1 has been described in other tissues. These observations let us to hypothesize that the expression of the protein ODF1 could not be limited to the testis. Therefore, in the present work we proposed to evaluate if the ODF1 protein could also be present in tissues other than the testis. Here we demonstrated through western blot, immunofluorescence, and RT-PCR techniques that the protein and mRNA of ODF1 have been identified in the rat kidney. Finally, the presence of ODF1 in kidney has also been confirmed through proteomic analysis using mass spectrometry. The results derived from these different complementary approaches indicate that, to our knowledge and for the first time, ODF1 is demonstrated to be present in an additional organ different to testis. This results raise new questions about potential other functions and locations of the ODF1 protein.

2.
Heliyon ; 5(11): e02868, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31844747

ABSTRACT

A chronic-positive energetic balance has been directly correlated with infertility in men, but the involved mechanisms remain unknown. Herein we investigated weather in a mouse model a chronic feeding with a diet supplemented with chicken fat affects sperm head morphology. To accomplish this, we fed mice for 16 weeks with either control food (low-fat diet, LFD) or control food supplemented with 22% chicken fat (high-fat diet, HFD). At the end of the feeding regimen, we measured: redox and inflammatory changes, cholesterol accumulation in testis and analyzed testicular morphological structure and ultra-structure and liver morphology. We found that the mice fed HFD resembled some features of the human metabolic syndrome, including systemic oxidative stress and inflammation, this group showed an increment in the following parameters; central adiposity (adiposity index: 1.07 ± 0.10 vs 2.26 ± 0.17), dyslipidemia (total cholesterol: 153.3 ± 2.6 vs 175.1 ± 8.08 mg/dL), insulin resistance (indirect Insulin resistance index, TG/HDL-c: 2.94 ± 0.33 vs 3.68 ± 0.15) and fatty liver. Increased cholesterol content measured by filipin was found in the testicles from HFD (fluorescence intensity increase to 50%), as well as an alteration of spermiogenesis. Most remarkably, a disorganized manchette-perinuclear ring complex and an altered morphology of the sperm head were observed in the spermatozoa of HFD-fed mice. These results add new information to our understanding about the mechanisms by which systemic oxidative stress and inflammation may influence sperm-head morphology and indirectly male fertility.

3.
Contrast Media Mol Imaging ; 8(5): 409-16, 2013.
Article in English | MEDLINE | ID: mdl-23740811

ABSTRACT

We used [(18)F]-4-fluorobenzamido-N-ethylamino-maleimide ([(18)F]-FBEM) to radiolabel cells ex vivo for in vivo positron emission tomography (PET) in order to assess cell trafficking in mice. In contrast to commonly used imaging agents, [(18)F]-FBEM forms a covalent bond with thiol groups present on the cells surface. The stability of the probe in aqueous medium was tested at different pH values and cross-experiment showed that thiol-labeling efficiency was retained (at least) up to pH 9. The labeling procedure did not affect significantly the cell viability. To illustrate the procedure, PET images of living mice injected intravenously with labeled T lymphocytes were obtained. They showed the expected cell homing in the spleen that was absent in mice injected with free label.


Subject(s)
Cell Tracking , Maleimides , Sulfhydryl Compounds/isolation & purification , T-Lymphocytes/ultrastructure , Animals , Cell Line, Tumor , Fluorine Radioisotopes , Maleimides/administration & dosage , Maleimides/chemistry , Mice , Mice, Nude , Positron-Emission Tomography , Radioisotopes , Staining and Labeling , Sulfhydryl Compounds/chemistry , Surface Properties , T-Lymphocytes/chemistry , Tissue Distribution
4.
Contrast Media Mol Imaging ; 1(5): 212-20, 2006.
Article in English | MEDLINE | ID: mdl-17193698

ABSTRACT

Labeled amino acids (AA) are tumor tracers for use in nuclear medecine. O-(2-[(18)F]fluoroethyl)-L-tyrosine (FET) is transported by the L-system, known to function as an exchanger. In vitro utilization of FET, after a preload or prior to an afterload of non radioactive L-amino acids, was evaluated in order to measure the potential effects of AA content on the distinction between tumor and inflammatory lesions. Cellular uptake of FET was studied on rat osteosarcoma cells (ROS 17/2.8) and human leukocytes, initially loaded with nonradioactive L-tyrosine or L-methionine. FET efflux was evaluated from cells loaded with nonradioactive L-phenylalanine after tracer uptake. ROS 17/2.8 showed a higher sensitivity to preload and afterload effects on cellular FET content as compared with the leukocytes. We conclude that preload with L-tyrosine, prior to the administration of FET, may be a potential procedure to improve PET differentiation between tumor and inflammatory lesions.


Subject(s)
Amino Acids/metabolism , Fluorodeoxyglucose F18/pharmacokinetics , Inflammation/diagnosis , Neoplasms/diagnosis , Positron-Emission Tomography/methods , Tyrosine/analogs & derivatives , Animals , Diagnosis, Differential , Humans , Leukocytes/metabolism , Methionine/metabolism , Phenylalanine/metabolism , Rats , Tyrosine/chemical synthesis , Tyrosine/pharmacokinetics
5.
Reproduction ; 129(3): 291-8, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15749956

ABSTRACT

Bovine sperm protease, 66 kDa (BSp66) is a serine protease previously characterized in bovine spermatozoa. Like other proteases, it may be present in sperm from other mammalian species different from bovine, playing a role in the fertilization process. In this study, we looked for BSp66 in hamster spermatozoa using heterologous antibodies against bovine BSp66. An immunoreactive protein was detected by Western blotting in mature and immature sperm. The detected protein had two isoforms similar to the ones reported in bovine sperm. Furthermore, indirect immune detection by fluorescence and electron microscopy assays, showed BSp66 signal at the acrosomal region similar to bovine sperm. As it was determined in bovine sperm, the acrosomal reaction displays the antigen within the acrosomal content. When live hamster sperm was incubated with polyclonal antibody against bovine BSp66 a decrease in the number of sperm bound to zona pellucida in homologous IVF and an impairment of head-head agglutination, were observed. These results suggest that a protease homologous to bovine BSp66 is present in golden hamster spermatozoa, with a conserved molecular mass and cellular location. Moreover, hamster BSp66 is probably involved in zona pellucida recognition.


Subject(s)
Cricetinae/metabolism , Serine Endopeptidases/metabolism , Sperm-Ovum Interactions/physiology , Spermatozoa/metabolism , Animals , Cattle , Cell Culture Techniques , Cell Survival , Fertilization in Vitro , Immunohistochemistry/methods , Male , Microscopy, Immunoelectron , Sperm Motility/physiology
6.
Biochem Biophys Res Commun ; 324(2): 874-7, 2004 Nov 12.
Article in English | MEDLINE | ID: mdl-15474509

ABSTRACT

Fertilization in mammals comprises a sequence of events leading to the fusion of sperm and oocyte membranes. Although proteases are known to be involved in this process, their role in fertilization is controversial. There is extensive work on the characterization of proteolytic systems, including serine proteases, which demonstrates that acrosomal proteases can be distinguished among the sperm of different mammalian species on the basis of the gelatin-hydrolyzing activity on SDS-PAGE by the quantity and variety of the enzymes. In this report, we investigated the occurrence and activity of the serine protease BSp66, previously characterized in bovine spermatozoa, in various mammalian sperm. A protein with a molecular mass of 66 kDa cross-reacted with heterologous antibodies against bovine BSp66 when sperm extracts of several mammalian species were analyzed by Western blot. In agreement, proteolytic activity corresponding to the molecular mass of BSp66 was detected by gelatin zymography in all the species analyzed. This protein was located on the acrosomal region of sperm cells by immunofluorescence methods. We concluded that BSp66 is widespread in mammalian sperm, with a conserved location in the acrosomal region.


Subject(s)
Acrosome/metabolism , Seminal Plasma Proteins/chemistry , Serine Endopeptidases/chemistry , Spermatozoa/metabolism , Animals , Blotting, Western , Cattle , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique, Indirect , Gelatin/metabolism , Hydrolysis , Male , Microscopy, Fluorescence , RNA, Messenger/metabolism , Species Specificity
7.
Rev Med Brux ; 23 Suppl 2: 101-5, 2002.
Article in French | MEDLINE | ID: mdl-12584923

ABSTRACT

During the last 25 years, the clinical and experimental activity in nuclear medicine at Erasme hospital has been influenced by the implementation of positron emission tomography (PET) in 1990 as a method of brain functional investigation. The activity of the PET/biomedical cyclotron unit has been dedicated to various subjects in neurology, neurosciences, psychiatry, oncology and cardiology. This has been made possible by developments in radiochemistry. The radiochemistry laboratory has designed and produced original tracers such as 9-[(3-[18F]fluoro-1-hydroxy-2-propoxy)-methyl]guanine (FHPG), a tracer of viral thymidine kinase activity in gene therapy protocols. We have brought new applications of PET, such as its integration into stereotactic neurosurgical and radioneurosurgical techniques in order to improve their diagnostic and therapeutic performance in neurooncology. We have also conducted multiple studies on brain physiology and pathophysiology, in particular with the use of functional and metabolic brain mapping methods and the use of tracers of neurotransmission systems. The Department of nuclear medicine has also performed studies on bone metabolism and investigated in vivo imaging methods of infectious and immune processes.


Subject(s)
Nuclear Medicine Department, Hospital , Belgium , Biomedical Research , Cyclotrons , Hospitals, University , Humans , Tomography, Emission-Computed
9.
J Neurosurg ; 95(5): 746-50, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11702862

ABSTRACT

OBJECT: Positron emission tomography with L-[methyl-11C]methionine (MET-PET) provides information on the metabolism of gliomas. The aim of this study was to determine the predictive value of MET-PET in the treatment of patients with gliomas. METHODS: Since 1992, 85 patients with a World Health Organization (WHO) classification-verified glioma underwent PET studies in which MET was injected before (74 cases) or after treatment (11 cases). Analysis of PET data was conducted by the same investigator using two scales: a qualitative visual grading scale and a quantitative scale (ratio between tumor uptake and normal brain uptake, classified on a seven-level scale). Uptake of MET was present in 98% of gliomas. The investigator judged this uptake to be moderate to very high based on visual inspection (qualitative scale). For all grades of gliomas, a visual grade of 3 was statistically associated with a shorter patient survival period (p < 0.005). The tumor/normal brain uptake ratio was significantly influenced by the histological grade of the tumor. A statistically poor outcome was demonstrated when this ratio was higher than a threshold of 2.2 for a WHO Grade II tumor and 2.8 for WHO Grade III tumor. For Grade II and III tumors, oligodendrogliomas had a higher uptake of MET than astrocytomas. CONCLUSIONS: Uptake of MET was present in 98% of the gliomas studied. A high uptake is statistically associated with a poor survival time. The intensity of MET uptake represents a prognostic factor for WHO Grade II and III tumors considered separately.


Subject(s)
Brain Neoplasms/diagnostic imaging , Glioma/diagnostic imaging , Methionine , Tomography, Emission-Computed , Adult , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Female , Glioma/metabolism , Glioma/pathology , Humans , Injections, Intravenous , Male , Methionine/pharmacokinetics , Prognosis , Survival Analysis
11.
Acta Neurol Belg ; 97(3): 183-6, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9345590

ABSTRACT

The first applications of positron emission tomography (PET) for the study of brain tumours appeared early in the development of this technology. New trends in these particular PET applications tend to take into account the histological heterogeneity of these tumours and the necessity to integrate PET data in their surgical management. Better knowledge on PET tracers behavior in brain tumours should lead to new clinical uses, in particular in the promising field of cancer treatment evaluation.


Subject(s)
Brain Neoplasms/diagnostic imaging , Tomography, Emission-Computed , Fluorine Radioisotopes , Fluorodeoxyglucose F18 , Glioma/diagnostic imaging , Humans , Prognosis , Treatment Outcome
12.
Nucl Med Biol ; 24(4): 357-60, 1997 May.
Article in English | MEDLINE | ID: mdl-9257335

ABSTRACT

To further validate its use in positron emission tomography (PET), we studied the binding of [18F]altanserin, a specific 5HT2 radioligand, in the rat brain using in vivo autoradiography. Distribution of [18F]altanserin binding was comparable to the in vitro mapping of 5HT2 receptors reported in the literature. Selective displacers were used to test the reversibility and the selectivity of this radioligand. Specific binding of [18F]altanserin in the rat frontal cortex was quantified by direct counting with an electronic imaging system and by quantification on digitalized autoradiograms. Close results of about 30 pmol/g were obtained with both methods. Our data confirmed that [18F]altanserin is a valid tracer for 5HT2 receptors binding studies.


Subject(s)
Brain/metabolism , Fluorine Radioisotopes , Ketanserin/analogs & derivatives , Receptors, Serotonin/analysis , Serotonin Antagonists/metabolism , Animals , Autoradiography , Ketanserin/metabolism , Male , Rats , Rats, Wistar , Receptors, Serotonin/metabolism
13.
Nucl Med Biol ; 23(4): 413-6, 1996 May.
Article in English | MEDLINE | ID: mdl-8832695

ABSTRACT

An automatic system allowing repetitive bolus injection of oxygen-15-labeled water for PET studies is described in this report. The production of this radiopharmaceutical by the 16O(p,pn)15O nuclear reaction on H2(16O), its purification and delivery nearby the PET camera, the injection system, and the quality controls are presented.


Subject(s)
Oxygen/administration & dosage , Oxygen/chemistry , Water/administration & dosage , Water/chemistry , Humans , Injections, Intravenous , Oxygen Isotopes , Tomography, Emission-Computed/methods
14.
Neurosci Lett ; 204(1-2): 25-8, 1996 Feb 02.
Article in English | MEDLINE | ID: mdl-8929969

ABSTRACT

Serotonergic mechanisms are involved in gender-related behaviors and psychiatric conditions like aggression, eating disorders, depression, impulsivity or suicide. We studied gender differences in the living human brain type-2 serotonin receptor (5HT2r). Twenty-two healthy age-matched men and women were investigated using positron emission tomography and the selective radiotracer, 18F-labeled altanserin. Binding was quantified using a non-linear least-squares minimization procedure. We found significantly higher 5HT2r binding capacity in men than in women, especially in the frontal and cingulate cortices. Distinct liability for men and women to suffer from some psychiatric disorders responding to serotonergic agents may be related to differences in brain serotonin receptors.


Subject(s)
Brain Chemistry/physiology , Receptors, Serotonin/metabolism , Adult , Female , Fluorine Radioisotopes , Humans , Ketanserin/analogs & derivatives , Ketanserin/pharmacokinetics , Male , Middle Aged , Sex Characteristics , Tomography, Emission-Computed
15.
Int J Biol Markers ; 10(3): 149-55, 1995.
Article in English | MEDLINE | ID: mdl-8551057

ABSTRACT

Various authors have proposed the use of hyaluronic acid (HA) as a tumor marker. In order to analyze its usefulness as a marker in bronchogenic carcinoma, the most common carcinoma in men, we determined the HA values in serum and bronchoalveolar lavage fluid (BAL). We performed prospective studies on two groups of patients: 81 diagnosed as having bronchial carcinoma and 34 with benign respiratory diseases. HA values were higher in patients with cancer than in those with benign diseases (serum: 79.8 ng/ml vs 63.7 ng/ml; BAL: 927 ng/mg vs 522 ng/mg). Also, the percentage of patients with levels exceeding the established cutoff was greater in the group with cancer than in the group with benign disease (serum: 24.6 vs 17.6; BAL: 25.3 vs 3). Statistically significant differences in these percentages were found in BAL (p<0.01). Patients with extended small cell carcinoma had higher HA values (p =0.04) than those with limited disease, and the percentage of patients with abnormal HA values was larger in the group with extended disease than in the group with limited disease (p = 0.004). The serial determinations of HA values in serum reflected the clinical evolution after treatment in 73% of the small cell carcinomas. Most of the patients with benign diseases whose HA values exceeded the cutoff level suffered from acute infectious dis-eases. Once these cases were excluded, the specificity of HA value determination in the diagnosis of carcinoma was very high (serum 96%, BAL 100%). The determination of HA levels in serum or BAL did not have any prognostic value in this study. We conclude that the HA levels in serum and BAL could be of interest as a tumor marker, especially in patients with small cell carcinoma.


Subject(s)
Biomarkers, Tumor/blood , Biomarkers, Tumor/metabolism , Bronchoalveolar Lavage Fluid/chemistry , Carcinoma, Bronchogenic/blood , Carcinoma, Bronchogenic/metabolism , Hyaluronic Acid/blood , Hyaluronic Acid/metabolism , Lung Neoplasms/blood , Lung Neoplasms/metabolism , Adenocarcinoma/blood , Adenocarcinoma/diagnosis , Adenocarcinoma/metabolism , Adult , Aged , Aged, 80 and over , Carcinoma, Bronchogenic/diagnosis , Carcinoma, Large Cell/blood , Carcinoma, Large Cell/diagnosis , Carcinoma, Large Cell/metabolism , Carcinoma, Small Cell/blood , Carcinoma, Small Cell/diagnosis , Carcinoma, Small Cell/metabolism , Carcinoma, Squamous Cell/blood , Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/metabolism , Case-Control Studies , Female , Humans , Lung Neoplasms/diagnosis , Male , Middle Aged , Prognosis , Prospective Studies , Respiratory Tract Diseases/blood , Respiratory Tract Diseases/metabolism
16.
Eur J Nucl Med ; 21(9): 937-46, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7995287

ABSTRACT

Serotoninergic type 2 (5HT2) receptors have been implicated in the regulation of many brain functions in humans and may play a role in several neurological and psychiatric diseases. Fluorine-18 altanserin has been proposed as a new radiotracer for the study of 5HT2 receptors by PET because of its high affinity for 5HT2 receptors (Ki: 0.13 nM) and its good specificity in in vitro studies. Dynamic PET studies were carried out in 12 healthy volunteers after intravenous injection of 0.1 mCi/kg [18F]altanserin. Ninety minutes after injection, we observed mainly cortical binding. Basal ganglia and cerebellum showed very low uptake and the frontal cortex to cerebellum ratio was about 3. To evaluate the quantitative distribution of this ligand in the brain, we used two different methods of data analysis: a four-compartment model was used to achieve quantitative evaluation of rate constants (K1 and k2 through k6) by non-linear regression, and a multiple-time graphical analysis technique for reversible binding was employed for the measurement of k1/k2 and k3/k4 ratios. Using both methods, we found significant differences in binding capacity (estimated by k3/k4 = Bmax/Kd) between regions, the values increasing as follows: occipital, limbic, parietal, frontal and temporal cortex. After correction of values obtained by the graphical method for the existence of non-specific binding, results generated by the two methods were consistent.


Subject(s)
Brain/diagnostic imaging , Fluorine Radioisotopes , Ketanserin/analogs & derivatives , Receptors, Serotonin/analysis , Tomography, Emission-Computed , Adult , Female , Humans , Male
17.
Eur J Pharmacol ; 257(1-2): 53-8, 1994 May 12.
Article in English | MEDLINE | ID: mdl-8082707

ABSTRACT

In the presence of carbidopa, L-3,4-dihydroxy-6-[18F]fluorophenylalanine ([18F]fluoro-DOPA) is mainly metabolized by catechol-O-methyl transferase. We studied the effects of entacapone, a peripheral catechol-O-methyl transferase inhibitor, on striatal [18F]fluoro-DOPA uptake in rats. Rats were pretreated with carbidopa, entacapone or both before high specific activity (> 2 Ci/mmol) [18F]fluoro-DOPA administration. Entacapone alone antagonized the appearance of methylated metabolites in plasma, striatum and cerebellum but did not increase striatal [18F]fluoro-DOPA availability. Entacapone added to carbidopa significantly increased the striatum/cerebellum total radioactivity ratio (1.4 versus 1.2 in rats with carbidopa, 1.0 in controls) but significant levels of methylated metabolites were found in the brain. Entacapone added to carbidopa might increase the striatum/cerebellum total radioactivity ratio in humans undergoing [18F]fluoro-DOPA positron emission tomography (PET) studies. However, the appearance of methylated metabolites in the brain could hamper quantification of the PET data.


Subject(s)
Catechol O-Methyltransferase Inhibitors , Cerebellum/metabolism , Corpus Striatum/metabolism , Dihydroxyphenylalanine/analogs & derivatives , Animals , Carbidopa/pharmacology , Catechols/administration & dosage , Catechols/pharmacology , Cerebellum/drug effects , Chromatography, High Pressure Liquid , Corpus Striatum/drug effects , Dihydroxyphenylalanine/blood , Dihydroxyphenylalanine/metabolism , Fluorine Radioisotopes , Male , Nitriles , Rats , Rats, Wistar
18.
Proc Natl Acad Sci U S A ; 85(15): 5597-600, 1988 Aug.
Article in English | MEDLINE | ID: mdl-16593967

ABSTRACT

North America and South America have recently been colonized by the Palearctic species Drosophila subobscura. This double colonization offers a rare opportunity for evolutionary studies. Correlations between chromosomal arrangement frequencies and latitude were calculated for the colonizing populations. Signs of these correlations are highly coincident with those found in the Old World. These results provide experimental support for the adaptive value of the chromosomal-inversion polymorphism; historical and other nonadaptive explanations are thus excluded or relegated to a secondary role.

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