BBS4 protein has basal body/ciliary localization in sensory organs but extra-ciliary localization in oligodendrocytes during human development.

Bardet-Biedl syndrome protein 4 (BBS4) localization has been studied in human embryos/fetuses from Carnegie stage 15 to 37 gestational weeks in neurosensory organs and brain, underlying the major clinical signs of BBS. We observed a correlation between the differentiation of the neurosensory cells (hair cells, photoreceptors, olfactory neurons) and the presence of a punctate BBS4 immunostaining in their apical cytoplasm. In the brain, BBS4 was localized in oligodendrocytes and myelinated tracts. In individual myelinated fibers, BBS4 immunolabelling was discontinuous, predominantly at the periphery of the myelin sheath. BBS4 immunolabelling was confirmed in postnatal developing white matter tracts in mouse as well as in mouse oligodendrocytes cultures. In neuroblasts/neurons, BBS4 was only present in reelin-expressing Cajal-Retzius cells. Our results show that BBS4, a protein of the BBSome, has both basal body/ciliary localization in neurosensory organs but extra-ciliary localization in oligodendrocytes. The presence of BBS4 in developing oligodendrocytes and myelin described in the present paper might attribute a new role to this protein, requiring further investigation in the field of myelin formation.

GLUT1 expression in human papillomavirus-positive anogenital lesions.

BACKGROUND: GLUT1, an ubiquitous glucose transporter in the mammalian cells, is upregulated in many tumours, including human papillomavirus (HPV)-induced head and neck or cervical cancer. OBJECTIVE: To study in anogenital lesions whether or not GLUT1 expression correlates with genomic high-risk HPV integration, the first step in neoplastic transformation. METHODS: Forty-three HPV-positive biopsies positive for either low-risk or high-risk HPV were selected. Paraffin sections adjacent to those tested for the presence of HPV were processed for GLUT1 immunocytochemistry. GLUT1 expression was analysed by two histologists, blinded to HPV type and status and then compared with HPV typing results. RESULTS: Two main staining patterns were observed, either staining from the basal to the granular layer or staining of superficial layers only. The first staining pattern corresponded to lesions with high number of episomal HPV-positive nuclei. Superficial staining was observed in lesions with low number of episomal HPV nuclei or when high-risk HPV was integrated in the cell genome. CONCLUSION: Our results show that GLUT1 overexpression correlates with the number of episomally infected cells in the lesion, but not with the type (low or high risk) of HPV.

Simultaneous T1 and T2 quantification of the myocardium using cardiac balanced-SSFP inversion recovery with interleaved sampling acquisition (CABIRIA).

PURPOSE: To develop a novel sequence for simultaneous quantification of T1 and T2 relaxation times in the myocardium based on the transient phase of the balanced steady-state free precession. METHODS: A new prototype sequence, named "cardiac balanced-SSFP inversion recovery with interleaved sampling acquisition" (CABIRIA) was developed based on a single-shot bSSFP readout following an inversion pulse. With this method, T1 and T2 values can be calculated from the analysis of signal evolution. The scan duration for a single slice in vivo was 8 heartbeats, thus feasible in a breath-hold. The sequence was validated both in vitro by comparing it to conventional inversion recovery and multi-echo spin-echo methods and in 5 healthy volunteers by comparing it to the Modified Look-Locker Inversion Recovery (MOLLI) sequence and to a T2 quantification sequence based on multi-T2 -prepared bSSFP. RESULTS: The method showed good agreement with conventional methods for both T1 and T2 measurements (concordance correlation coefficient ≥ 0.99) in vitro. In healthy volunteers the measured T1 values were 1227 ± 68 ms and T2 values 37.9 ± 2.4 ms, with similar inter- and intrasubject variability with respect to existing methods. CONCLUSION: The proposed CABIRIA method enables simultaneous quantification of myocardial T1 and T2 values with good accuracy and precision.

Novel SPG10 mutation associated with dysautonomia, spinal cord atrophy, and skin biopsy abnormality.

BACKGROUND: SPG10 is a rare form of autosomic dominant hereditary spastic paraplegia (HSP) caused by mutations in the KIF5A gene, which may be involved in axonal transport. METHODS: We report the characteristics of a French family with a novel missense mutation c.580 G>C in exon 7 of the KIF5A gene. RESULTS: The proband and his sister presented with an adult onset HSP, a sensory spinal cord-like syndrome, dysautonomia, and severe axonal polyneuropathy. Contrary to the proband, his sister presented a secondary improvement in spasticity and walking. In the proband, MRI findings consisted in spinal cord atrophy and symmetric cerebral demyelination, whereas the skin biopsy suggested a defect in the number of vesicles and synaptophysin density at the pre-synaptic membrane. CONCLUSION: This study extends the phenotype of SPG10 and argues for abnormalities in the axonal vesicular transport.

[Peripheral neuropathies after bariatric surgery]. / Polyneuropathies après chirurgie bariatrique.

INTRODUCTION: Peripheral neuropathies sometimes complicate bariatric surgery. PATIENTS AND METHODS: We report the detailed clinical, electrophysiological, biological and histological characteristics of five patients who developed peripheral neuropathy after bariatric surgery. RESULTS: Three patients presented with small fiber neuropathy, one presented with axonal polyneuropathy, and one with demyelinating polyradiculoneuropathy. All patients had in common prominent neuropathic pain, massive weight loss, and multiple nutritional deficiencies. The pathophysiology of postbariatric surgery polyneuropathies is complex and involves nutritional, infectious and dysimmune mechanisms. CONCLUSION: The spectrum of peripheral neuropathies complicating bariatric surgery is wide, and includes pure small fiber neuropathy, axonal polyneuropathy, and demyelinating polyradiculoneuropathy. Treatment is mainly preventive, but sometimes surgical revision is needed.

[Confirmation of the use of skin biopsy in small-fiber neuropathy. First results]. / Confirmation de l'intérêt de la biopsie cutanée dans la neuropathie des petites fibres. Résultats préliminaires.

INTRODUCTION: In small-fiber neuropathy, skin biopsy reveals a reduction of intraepidermal nerve fiber density (IENFD), a feature often necessary for diagnosis. In France, this technique has not been widely used for this purpose. PATIENT AND METHOD: To validate this method, we studied 13 patients with suspected small-fiber neuropathy, analyzed their nervous intra- and subepidermal network with a punch skin biopsy and compared our data with those of literature. RESULTS: Ten patients had pure small-fiber neuropathy and three an axonal polyneuropathy involving large-caliber nerve fibers. In the group of patients with pure small-fiber neuropathy, we found medium IENFD (11.6 +/- 4.46 fibers per millimeter in the proximal thigh and 7.15 +/- 3.59 fibers per millimeter in distal leg), well correlated with the electron microscopy quantitative and qualitative analysis of the unmyelinated subepidermal fibers. CONCLUSION: This work demonstrated the good reproducibility of skin biopsy for analyzing the small-fibers in our cohort. These results require further confirmation in a larger cohort and validation in comparison with controls analyzed on a local level. Nevertheless, these techniques seem to be useful to assess the difficult diagnosis of small-fiber neuropathy.

[Functional histology of dermis]. / Histologie fonctionnelle du derme.

The skin is composed of epidermis, dermis and subcutaneous tissue that interconnect anatomically. The dermis is an integrated system of fibrous and amorphous connective tissue that accommodates nerve and vascular networks, epidermally derived appendages, fibroblasts, macrophages and mast cells. Elastic and collagen tissue are the main types of fibrous connective tissue. The elastic connective tissue is assembled in a continuous network including mature elastic fibers, immature elaunin fibers and oxytalan fibers. Mature elastic fibers and elaunin have microfibrillar and amorphous matrix components while oxytalan fibers only contain microfibrils. Several molecules have been identified as constituents of the elastic fibers. Among the most characterized of these molecules is elastin in amorphous matrix, fibrillins 1 and 2 and LTBP-2 (ligand of latent TGFbeta) in microfibrils and fibulins which interconnect elastin and fibrillins. Elastic fibers provides elasticity to the skin. Under electron microscope, collagen fibers appears as of bundles of periodically banded fibrils which are composed of collagens types I, III and V; type V collagen is believed to assist in regulating fibril diameter. They are associated with FACITs (fibril-associated collagen with interrupted triple helixes) collagens types XIV et XVI. Collagen fibers provide tensile strength to the skin. Non fibrous connective tissue molecules include finely filamentous glycoproteins, glycosaminoglycans and proteoglycans of "the ground substance" (hyaluronic acid and chondroitin sulphate, dermatan sulphate, versican, decorin). Fibroblasts, macrophages and mast cells are regular residents of the dermis. The main function of these cells are well known. Fibroblasts are responsible for the synthesis and the degradation of fibrous and non fibrous connective tissue matrix proteins. Macrophages are phagocytic; they process and present antigen to immunocompetent lymphoid cells. Mast cells are responsible for IgE mediated acute, subacute and chronic inflammation. All these cells have a long list of other functions, in particular they are involved in coagulation, wound healing and tissue remodeling.

Diagnosis of primary ciliary dyskinesia: When and how?

INTRODUCTION: Primary ciliary dyskinesia (PCD) is a rare congenital disorder involving permanent ubiquitous structural and/or functional ciliary abnormalities. METHODS: A single-center retrospective study included 56 cases of PCD (respiratory form) out of a cohort of 280 patients with suspected PCD. The main features of history-taking and clinical examination were analyzed, to formulate a pragmatic diagnostic procedure, easy to implement in clinical practice. RESULTS: Chronic respiratory tract infectious symptoms are sensitive but non-specific for the diagnosis of PCD. Nasal brushing for phase-contrast microscopy study of ciliary morphology and activity proved to be a fast, easy, non-invasive, cost-effective and age-independent diagnostic method. In doubtful cases, depending on local availability, further tests are indicated: nasal nitric oxide level, electronic microscopy, genetic study and cell culture. CONCLUSIONS: In suspected PCD, there being no gold standard method of screening and early diagnosis, nasal brushing with ciliary study is contributive, alongside numerous other complementary tests, on condition that the clinician is experienced and results are interpreted in the light of clinical examination and history-taking.

X-linked reticulate pigmentary layer. Report of a new patient and demonstration of a skewed X-inactivation.

We report a boy, born to healthy first cousin parents, with diffuse hyperpigmentation of the skin and guttate hypomelanotic lesions, photophobia, abnormal hair, developmental delay, and recurrent bronchitis. Skin histology showed pigmentation incontinence with numerous melanophages. Electron microscopy showed a very high number of melanosomes and some degenerating keratinocytes. These features correspond to a rare genodermatosis, the X-linked reticulate pigmentary disorder with systemic manifestations. Skewed X-inactivation patterns were detected in the mother's lymphocytes.

[Gene transfers of kallikrein, bradykinin receptor B2 and a mutated B2 receptor stimulate neoangiogenesis in peripheral ischemia]. / La surexpression de la kallicréine tissulaire, du récepteur B2 de la bradykinine et d'une forme mutée du récepteur B2, stimule la néoangiogenèse dans un modèle d'ischémie périphérique.

In this work, we evaluated the angiogenic effect of the gene transfer of human tissue kallikrein (TK), bradykinin B2 receptor (B2R) and a mutated form (RB2m) in a rabbit peripheral model of ischaemia. We studied the effects of the transfection of each of these factors and the effects of their co-transfection. In New Zealand anesthetised rabbits we first induced an ischaemia of the left posterior leg by ligation-excision of the superficial femoral artery and its collaterals. Seven days later, we performed i.m. injections in the ischemic tight with transfection solutions containing either the control (pcDNA3 empty backbone) or the pcDNA3-TK, the pcDNA3-TK and the pcDNA3-B2R, the pcDNA3-TK and the pcDNA3-B2Rm. Twenty eight days later, the therapeutic effect was evaluated using ultrasonographic debitmetry of the common iliac artery, perfusion index (PI) = ischemic leg blood flow /non ischemic leg blood flow (%) and capillaries measurements i.e. capillary density: number of vessels/mm2 and the ratio of vessels/muscular fiber, in the adductors and gastrocnemian muscles. The PI was increased in each treated group vs control (32.61 +/- 5.2%), pcDNA3-TK: 59.72 +/- 2.33%; p = 0.001; pcDNA3-RB2: 55.25 +/- 2.29%; p = 0.008; pcDNA3-TK + pcDNA3-RB2: 84.77 +/- 3.15%; p < 0.001; pcDNA3-TK + pcDNA3-RB2m: 103.25 +/- 4.9%; p < 0.001. The capillary density and the vessel/muscular fiber ratio increased in a parallel with the hemodynamic in the ischemic adductors (pcDNA3-TK + pcDNA3-B2Rm > pcDNA3-TK + pcDNA3-RB2 > pcDNA3-TK = pcDNA3-B2R; p < 0.001). There was no angiogenic effect measurable neither in the non ischemic adductors (right) nor in the gastrocnemian muscles. In rabbit peripheral ischaemia, the cotransfection of TK and B2R increases the arterial flow in the treated leg and potentiates the neoangiogenesis. Cotransfection of the B2Rm cDNA enhanced the synergic effect of this therapeutic strategy.

Retinoid receptors in rat vaginal and uterine epithelia: changes with ovarian steroids.

In rats, vaginal epithelium shows cyclic changes with an alternating pattern of keratinization under estrogen control and mucification under progesterone control. Since retinoids suppress keratinizing differentiation, in this paper we studied the expression of the major retinoid receptors normally present in keratinizing squamous epithelia: RAR alpha, RAR gamma and RXR alpha. In cyclic rats and steroid-treated ovariectomized rats, RXR alpha and RAR gamma were detected in basal and suprabasal cells while RAR alpha was mainly localized in suprabasal cells. No changes in RAR gamma expression were observed in correlation with ovarian steroids. During diestrus and in ovariectomized rats, the superficial cuboid cells expressed the three receptors. In the uterine epithelium, RAR alpha, RAR gamma and RXR alpha expression was induced by estrogens. Retinoic acid treatment did not modify retinoid receptor expression in vaginal and uterine epithelia. These data suggest specific roles for the different receptors in the complex process of vaginal epithelium proliferation and differentiation under estrogens and retinoic acid control.

Sensory receptor-like cells in the human foetal vomeronasal organ.

The vomeronasal organ (VNO) was studied in ten human foetuses, 12 to 36 weeks old. At 12 to 23 weeks, it was lined by a smooth pseudostratified epithelium, with neurone-specific enolase (NSE) positive cells looking like olfactory receptors. Clusters of NSE-positive cells were seen in relation with the posterosuperior end of the organ and along nerve fascicles in the nasal septum. At 36 weeks, the organ was lined by a respiratory epithelium and did not show any receptor-like cells; some pear-shaped NSE-positive cells of unknown significance were seen at the upper part of the respiratory epithelium. Our results suggest that during the early foetal period, the VNO could have some, as yet unknown, sensory function.

Identification of human papillomavirus in cervical swabs by deoxyribonucleic acid in situ hybridization.

A series of 47 lesions diagnosed cytologically as cervical intraepithelial neoplasia (CIN) III in 22 cases, CIN I/II in 13 cases, and 12 cases showing abnormal smears consistent with human papillomavirus infection were analyzed. Thirty-six cases with negative cytology were used as the control group. Sixty-eight percent of CIN III were positive for a mixture of human papillomavirus 16 and human papillomavirus 18, 18% reacted with human papillomavirus 6 or 11, and 14% were negative. Of the group with CIN I/II or with abnormal Papanicolaou smears, approximately one-third contained human papillomavirus 6 (11) and one-third human papillomavirus 16 and 18. Only 11% of the samples from the control group hybridized with human papillomavirus 6 (11), the others were negative with either probe. The data obtained by the rapid in situ hybridization of cervical cells are in agreement with the presence of human papillomavirus 16 and 18 in a high proportion of cervical carcinoma and carcinoma in situ lesions. Thus, the method can be applied to test the hypothesis that a lesion containing human papillomavirus 16/18 positive cells has a higher risk of progressing to cancer than a lesion harboring human papillomavirus 6 or 11.

Ontogenesis of NADPH-diaphorase activity in the olfactory bulb of the rat.

The enzyme NADPH-diaphorase, which has been shown to correspond to nitric oxide synthase, is present in discrete neuron populations in the olfactory bulb of the adult rat. The ontogenesis of NADPH-diaphorase activity was studied and compared with the ontogenesis of tyrosine hydroxylase containing cells from embryonic day E15 to postnatal day P30. In the main olfactory bulb, scanty NADPH-diaphorase reactive neurons were first present at E21 in an immature phenotype. The periglomerular positive cells increased in number and acquired their adult morphology in the postnatal period. No colocalization of tyrosine hydroxylase with NADPH-diaphorase was observed at any developmental stage studied. In the granule cell layer, a population of rather bipolar neurons transiently expressed NADPH-diaphorase from P3 to P15; a population of large multipolar cells permanently expressed NADPH-diaphorase from P3 to P30. In the accessory olfactory bulb, NADPH-diaphorase staining appeared in the granule cell layer at P3, and then in the granule cell projections towards the mitral cells. From E21 to P7, neural processes often seemed to contact blood vessels. Endothelial cells showed a diffuse and faint staining at all stages; moreover patches of high NADPH-diaphorase staining were transiently present on blood vessels from E15 to P7. The presence of both permanent and transient expression of NADPH-diaphorase during olfactory bulb genesis is discussed according to the hypotheses of the function of NO during development.

Luteinizing hormone-releasing hormone (LHRH)-expressing cells in the nasal septum of human fetuses.

In 12-19 weeks old human fetuses, LHRH-immunoreactive (LHRH-Ir) cells were detected in the nerve fascicles arising from the vomeronasal organ (VNO), from their origin to their end point at the medial aspect of the olfactory bulb (vomeronasal and/or terminal nerves). In one 22-weeks-old fetus, LHRH-Ir cells were present only in the upper part of the nasal septum, in the nerve fascicles crossing the cribriform plate. No LHRH cells could be detected in two fetuses, 23 and 36 weeks old. Our results are discussed with regard to the migratory hypothesis of LHRH cells from the medial olfactory placode to the brain.

Expression of NADPH-diaphorase in the rat forebrain during development.

Expression of NADPH-diaphorase (NADPH-d) was studied in the rat telencephalon and diencephalon from embryonic day 15 (E15) to postnatal day 30 (P30). The study has focused on the first appearance of NADPH-d staining in some areas which show high expression during adult life. The time of appearance ranged from E15 to the first days following birth, depending on the location of the cells. In many regions, neuronal processes, when staining appeared, were observed in close relationship with cerebral vessels. A possible role for nitric oxide in brain development should be explored.

Facilitory effects of olfactory cues emitted by estrous females on mounting behavior in the female rat.

Adult ovariectomized rats were implanted with testosterone propionate-filled silastic capsules and exposed to the odor of urine collected from ovariectomized rats made estrous by estrogen treatment. When caged with highly receptive females these androgenized female rats displayed more frequent mounting behavior with pelvic thrusting than their nonexposed counterparts (25/31 vs. 20/39). Olfactory environment may then be considered as a modulatory factor of heterotypic sexual behavior in the female rat.

Inhibitory effects of centrally and peripherally induced anosmia on mounting behavior in the female rat.

Adult ovariectomized female rats received a testosterone-propionate (TP)-filled silastic implant and were tested for mounting behavior seven days later. Exposure to estrous female urine but not to urine from ovariectomized females increased the number of TP-treated females that displayed mounting behavior. Olfactory bulb removal on its own and peripheral anosmia induced by intranasal ZnSO4 application impaired mounting behavior. These results are discussed with respect to the effects of anosmia on copulatory behavior in the male.

Interactions of testosterone with the olfactory system in the display of mounting behavior in the female rat.

Adult ovariectomized female rats received a testosterone propionate (TP) silastic implant (day 0) and were subsequently tested for mounting behavior. The number of females displaying mounting behavior increased from day 2 (0%) to day 14 (94%) following testosterone implantation. Olfactory bulbectomy (OB) performed on day -7 or on day +14 reduced the number of females that mounted on day +16 as compared to sham counterparts. However, the number of females displaying mounting behavior was significantly higher when OB was performed on day +14 than on day -7. Peripheral anosmia induced by ZnSO4 instillation, either with multiple instillations beginning on day -8 before TP implant or with a single injection on day +14, equally impaired mounting behavior on day +16. These results, which further confirm the role of the olfactory system in facilitating the display of mounting behavior in the female rat, suggest an olfaction-testosterone interaction.