A Brazilian family with hereditary inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia

Inclusion body myopathy associated with Paget disease and frontotemporal dementia (IBMPFD) is a progressive and usually misdiagnosed autosomal dominant disorder. It is clinically characterized by a triad of features: proximal and distal myopathy, early onset Paget disease of bone (PDB), and frontotemporal dementia (FTD). It is caused by missense mutations in the valosin-containing protein (VCP) gene. We describe here the clinical and molecular findings of the first Brazilian family identified with IBMPFD. Progressive myopathy affecting the limb girdles was detected by clinical examination followed by muscle biopsy and creatine kinase measurement. PDB was suggested after anatomopathological bone examination and FTD was diagnosed by clinical, neuropsychological and language evaluations. Brain magnetic resonance revealed severe atrophy of the anterior temporal lobes, including the hippocampi. A R93C mutation in VCP was detected by direct sequencing screening in subject W (age 62) and in his mother. Four more individuals diagnosed with "dementia" were reported in this family. We also present a comprehensive genotype-phenotype correlation analysis of mutations in VCP in 182 patients from 29 families described in the literature and show that while IBM is a conspicuously penetrant symptom, PDB has a lower penetrance when associated with mutations in the AAAD1 domain and FTD has a lower penetrance when associated with mutations in the Junction (L1-D1) domain. Furthermore, the R93C mutation is likely to be associated with the penetrance of all the clinical symptoms of the triad.

Streptococcus mutans GlnK protein: an unusual PII family member

Streptococcus mutans is a Gram-positive bacterium present in the oral cavity, and is considered to be one of the leading causes of dental caries. S. mutans has a glnK gene, which codes for a PII-like protein that is possibly involved in the integration of carbon, nitrogen and energy metabolism in several organisms. To characterize the GlnK protein of S. mutans, the glnK gene was amplified by PCR, and cloned into the expression vectors pET29a(+) and pET28b(+). The native GlnK-Sm was purified by anion exchange (Q-Sepharose) and affinity (Hi-Trap Heparin) chromatography. The GlnK-His-Sm protein was purified using a Hi-Trap Chelating-Ni2+ column. The molecular mass of the GlnK-His-Sm proteins was 85 kDa as determined by gel filtration, indicating that this protein is a hexamer in solution. The GlnK-His-Sm protein is not uridylylated by the Escherichia coli GlnD protein. The activities of the GlnK-Sm and GlnK-His-Sm proteins were assayed in E. coli constitutively expressing the Klebsiella pneumoniae nifLA operon. In K. pneumoniae, NifL inhibits NifA activity in the presence of high ammonium levels and the GlnK protein is required to reduce the inhibition of NifL in the presence of low ammonium levels. The GlnK-Sm protein was unable to reduce NifL inhibition of NifA protein. Surprisingly, the GlnK-His-Sm protein was able to partially reduce NifL inhibition of the NifA protein under nitrogen-limiting conditions, in a manner similar to the GlnK protein of E. coli. These results suggested that S. mutans GlnK is functionally different from E. coli PII proteins.

Players of Th-cell differentiation and immune dysregulation – focus on GIMAP family genes

The balance of T helper (Th) cell differentiation is the fundamental process that ensures that the immune system functions correctly and effectively. The differentiation is a fine tuned event, the outcome of which is driven by activation of the T-cell in response to recognition of the specific antigen presented. The co-stimulatory signals from the surrounding cytokine milieu help to determine the outcome. An impairment in the differentiation processes may lead to an imbalance in immune responses and lead to immune-mediated pathologies. An over-representation of Th1 type cytokine producing cells leads to tissue-specific inflammation and autoimmunity, and excessive Th2 response is causative for atopy, asthma and allergy. The major factors of Th-cell differentiation and in the related disease mechanisms have been extensively studied, but the fine tuning of these processes by the other factors cannot be discarded. In the work presented in this thesis, the association of T-cell receptor costimulatory molecules CTLA4 and ICOS with autoimmune diabetes were studied. The underlying aspect of the study was to explore the polymorphism in these genes with the different disease rates observed in two geographically close populations. The main focus of this thesis was set on a GTPase of the immunity associated protein (GIMAP) family of small GTPases. GIMAP genes and proteins are differentially regulated during human Th-cell differentiation and have been linked to immune-mediated disorders. GIMAP4 is believed to contribute to the immunological balance via its role in T-cell survival. To elucidate the function of GIMAP4 and GIMAP5 and their role in human immunity, a study combining genetic association in different immunological diseases and complementing functional analyses was conducted. The study revealed interesting connections with the high susceptibility risk genes. In addition, the role of GIMAP4 during Th1-cell differentiation was investigated. A novel function of GIMAP4 in relation to cytokine secretion was discovered. Further assessment of GIMAP4 and GIMAP5 effect for the transcriptomic profile of differentiating Th1-cells revealed new insights for GIMAP4 and GIMAP5 function.

Family rivalry in the testis: Retinoblastoma protein and E2F transcription factors in testicular development and adult spermatogenesis

Disorders of male reproductive health are becoming increasingly prevalent globally. These defects, ranging from decreasing sperm counts to an increasing rate of infertility and testicular cancer, have a common origin in the early phases of testicular development, but the exact mechanisms that cause them remain unknown. Testicular development and adult spermatogenesis are complex processes in which different cell types undergo mitosis, meiosis, differentiation and apoptosis. The retinoblastoma protein family and its associated E2F transcription factors are key regulators of these cellular events. In the present study, the functions of these factors in postnatal testicular development and adult spermatogenesis were explored using different animal models. In addition, a new application of flow cytometry to study testicular cell dynamics was developed. An ablation of retinoblastoma protein in mouse Sertoli cells resulted in their cell cycle re-entry in adult testes, dedifferentiation and a severe spermatogenic defect. We showed that deregulated E2F3 contributed to these changes. Our results indicated that the E2F1 transcription factor is critical for the control of apoptosis in the developing postnatal testis. In the adult testis, E2F1 controls the maintenance of the spermatogonial stem cell pool, in addition to inhibiting apoptosis of spermatocytes. In summary, this study elucidated the complex interdependencies of the RB and E2F transcription factor families in the control of postnatal testicular development and adult spermatogenesis. Furthermore, this study provided a new methodology for the analysis of testicular cells.

Ambassades en Espagne de Sébastien de L'Aubespine, évêque de Limoges, de Raymond de Pavie, baron de Forquevaulx, et du cardinal Louis de Guise ; originaux et copies ; pièces diverses. (1561-1572).

Contient : I ; Registre de l'ambassade de Sébastien de L'Aubespine, évêque de Limoges, en Espagne ; dépêches et mémoires de l'ambassadeur et pièces diverses, XVIe siècle, avec quelques parties du XVIIe, juillet 1561-mai 1562 ; On remarque : Contrat de mariage d'Élisabeth de France, 1559, et pièces annexes (f. 22), — et « Mémoire baillé à Sa Majesté Catholique, de ce qu'avoit esté pillé dans le gallion de Sa Majesté Très chrestienne », 1561 (f. 27) ; II ; Registre des dépêches adressées par la Cour à Mr de Forquevaulx et au cardinal de Guise, ambassadeurs en Espagne, et autres personnages, octobre 1567-juillet 1569 ; On remarque: « Instruction baillée à Mr de La Trimouille, allant en Espaigne, septembre 1568 » (f. 276 v), — et « Instruction au sr de Fourquevaulx..., ambassadeur près le Roy Catholicque..., de ce qu'il aura à dire... pour le faict du mariaige d'entre le Roy et madame Élisabeth, seconde fille de l'Empereur » (f. 300 v) ; III Pièces originales de l'ambassade de Mr de Forquevaulx en Espagne ; Nombreuses dépêches orig. de Mr de Forquevaulx à la Cour, septembre 1567-avril 1572, et minutes de lettres de la Cour [Charles IX, Catherine de Médicis], à Mr de Forquevaulx et à divers personnages (f. 315) ; Quelques lettres orig. de Gragnague, 1568 (f. 425), — de Loys, cardinal de Guyse, envoyé en Espagne « pour se conduloir de la mort de la reyne d'Espagne », Élisabeth de France, 1568-1569 (f. 480, etc.), — de Girolamo Gondi, 1569 (f. 497), — et de don Carlos, 1569 (f. 499, etc.) ; « Relation de l'arrivée du cardinal de Guyse à Madrid et de ce qu'il y a négotié, depuis le 28 janvier [1569]..., jusques au 9 février » ; Pièces diverses, parmi lesquelles on remarque: Copie d'une lettre du duc d'Albe, 1568 (f. 340), — « Mémoire des intentions du Roy, touchant le chastiment des huguenots de son royaulme », 1568 (f. 351), — Certificat orig. d'Andres de Salazar (f. 429), — et « Instruction donnée de la part du Roy au sr de La Trimouille, allant en Espagne, sur la mort du prince Carlos », minute (f. 439) ; Pièces relatives à l'ambassade du cardinal de Guise ; Copie d'une lettre de Philippe II au duc d'Albe ; « Discours d'un estranger dézireux de servir le Roy et s'employer vers les Indes », 1570 ; « Mémoire touchant douze François venans de la Floride, faicts prisonniers à Séville et condamnez aux gallaires pour dix ans »

Ambassades en Espagne de Sébastien de L'Aubespine, évêque de Limoges, de Raymond de Pavie, baron de Forquevaulx, et du cardinal Louis de Guise ; originaux et copies ; pièces diverses. (1561-1572).

Contient : I ; Registre de l'ambassade de Sébastien de L'Aubespine, évêque de Limoges, en Espagne ; dépêches et mémoires de l'ambassadeur et pièces diverses, XVIe siècle, avec quelques parties du XVIIe, juillet 1561-mai 1562 ; On remarque : Contrat de mariage d'Élisabeth de France, 1559, et pièces annexes (f. 22), — et « Mémoire baillé à Sa Majesté Catholique, de ce qu'avoit esté pillé dans le gallion de Sa Majesté Très chrestienne », 1561 (f. 27) ; II ; Registre des dépêches adressées par la Cour à Mr de Forquevaulx et au cardinal de Guise, ambassadeurs en Espagne, et autres personnages, octobre 1567-juillet 1569 ; On remarque: « Instruction baillée à Mr de La Trimouille, allant en Espaigne, septembre 1568 » (f. 276 v), — et « Instruction au sr de Fourquevaulx..., ambassadeur près le Roy Catholicque..., de ce qu'il aura à dire... pour le faict du mariaige d'entre le Roy et madame Élisabeth, seconde fille de l'Empereur » (f. 300 v) ; III Pièces originales de l'ambassade de Mr de Forquevaulx en Espagne ; Nombreuses dépêches orig. de Mr de Forquevaulx à la Cour, septembre 1567-avril 1572, et minutes de lettres de la Cour [Charles IX, Catherine de Médicis], à Mr de Forquevaulx et à divers personnages (f. 315) ; Quelques lettres orig. de Gragnague, 1568 (f. 425), — de Loys, cardinal de Guyse, envoyé en Espagne « pour se conduloir de la mort de la reyne d'Espagne », Élisabeth de France, 1568-1569 (f. 480, etc.), — de Girolamo Gondi, 1569 (f. 497), — et de don Carlos, 1569 (f. 499, etc.) ; « Relation de l'arrivée du cardinal de Guyse à Madrid et de ce qu'il y a négotié, depuis le 28 janvier [1569]..., jusques au 9 février » ; Pièces diverses, parmi lesquelles on remarque: Copie d'une lettre du duc d'Albe, 1568 (f. 340), — « Mémoire des intentions du Roy, touchant le chastiment des huguenots de son royaulme », 1568 (f. 351), — Certificat orig. d'Andres de Salazar (f. 429), — et « Instruction donnée de la part du Roy au sr de La Trimouille, allant en Espagne, sur la mort du prince Carlos », minute (f. 439) ; Pièces relatives à l'ambassade du cardinal de Guise ; Copie d'une lettre de Philippe II au duc d'Albe ; « Discours d'un estranger dézireux de servir le Roy et s'employer vers les Indes », 1570 ; « Mémoire touchant douze François venans de la Floride, faicts prisonniers à Séville et condamnez aux gallaires pour dix ans »

« Lettres de JEAN, cardinal DU BELLAY, escrittes à Charles de Lorraine, cardinal » de Guise. Minutes. Ces lettres, au nombre de 34, sont comprises entre le 9 janvier 1548 n. st. et le 15 janvier 1550 n. st. On trouve aussi dans ce volume.

Contient : 35 Minute d'une lettre « à monseigr de Guyse, le XIIIIe de juing 1550 », envoyée par « l'ordinaire » ; 36 « Memoyre à monseigneur le cardinal de Guyse, le 1er jour de febvrier », et addition audit mémoire