Signalementen

Het aantal valincidenten met ouderen kan tot de helft worden teruggebracht wanneer het gebruik van medicijnen zoals slaapmiddelen, kalmeringsmiddelen, antidepressiva of middelen tegen hart- en vaatziekten wordt verminderd of zelfs gestopt. Dit concludeert epidemioloog/ klinisch geriater i.o. Nathalie van der Velde, 32 jr, van het Erasmus MC in haar onderzoek waarop zij 23 mei 2007 promoveerde (proefschrift Falls in old age: pills, the heart and beyond, 139 p, ISBN 978 90 8559 297 2, promotores prof.dr. B.H.Ch. Stricker en prof.dr. H.A.P. Pols). Dertig procent van de 65-plussers in Nederland, oplopend tot vijftig procent van de 85-plussers, valt tenminste éénmaal per jaar en het was reeds bekend dat dit werd beïnvloed door bepaalde medicijnen.     

Comparative survival analysis of 12 histidine kinase mutants of Deinococcus radiodurans after exposure to DNA-damaging agents

Bacteria are able to adapt to changes in the environment using two-component signal transduction systems (TCSs) composed of a histidine kinase (HK) and a response regulator (RR). Deinococcus radiodurans, one of the most resistant organisms to ionizing radiation, has 20 putative HKs and 25 putative RRs. In this study, we constructed 12 D. radiodurans mutant strains lacking a gene encoding a HK and surveyed their resistance to γ-radiation, UV-B radiation (302 nm), mitomycin C (MMC), and H₂O₂. Five (dr0860 ^?, dr1174 ^?, dr1556 ^?, dr2244 ^?, and dr2419 ^?) of the 12 mutant strains showed at least a one-log cycle reduction in γ-radiation resistance. The mutations (1) dr1174, dr1227, and dr2244 and (2) dr0860, dr2416, and dr2419 caused decreases in resistance to UV radiation and MMC, respectively. Only the dr2416 and dr2419 mutant strains showed higher sensitivity to H₂O₂ than the wild-type. Reductions in the resistance to γ-radiation and H₂O₂, but not to UV and MMC, were observed in the absence of DR2415, which seems to be a cognate RR of DR2416. This result suggests that DR2415/DR2416 (DrtR/S: DNA damage response TCS) may be another TCS responsible for the extreme resistance of D. radiodurans to DNA-damaging agents.     

Direct Repeat 6 from Human Herpesvirus-6B Encodes a Nuclear Protein that Forms a Complex with the Viral DNA Processivity Factor p41

The SalI-L fragment from human herpesvirus 6A (HHV-6A) encodes a protein DR7 that has been reported to produce fibrosarcomas when injected into nude mice, to transform NIH3T3 cells, and to interact with and inhibit the function of p53. The homologous gene in HHV-6B is dr6. Since p53 is deregulated in both HHV-6A and -6B, we characterized the expression of dr6 mRNA and the localization of the translated protein during HHV-6B infection of HCT116 cells. Expression of mRNA from dr6 was inhibited by cycloheximide and partly by phosphonoacetic acid, a known characteristic of herpesvirus early/late genes. DR6 could be detected as a nuclear protein at 24 hpi and accumulated to high levels at 48 and 72 hpi. DR6 located in dots resembling viral replication compartments. Furthermore, a novel interaction between DR6 and the viral DNA processivity factor, p41, could be detected by confocal microscopy and by co-immunoprecipitation analysis. In contrast, DR6 and p53 were found at distinct subcellular locations. Together, our data imply a novel function of DR6 during HHV-6B replication.     

Ricerche biometriche sulla superficie filtrante del glomerulo renale di Rana esculenta

Con 4 figure nel testo. Al prof.Defrise spetta l'ideazione del tema e l'elaborazione biometrica statistica, al prof.Barelli l'esecuzione del rilievo microscopico quantitativo. Il testo venne steso di comune accordo.     

Improving Pseudomonas alcaligenes lipase’s diastereopreference in hydrolysis of diastereomeric mixture of menthyl propionate by site-directed mutagenesis

The resolutions of racemic diastereomeric mixtures of menthyl propionate was performed by Pseudomonas alcaligenes lipase (PaL) to produce (2S, 5R) L-menthol. Because of the inherently low diastereopreference of PaL, covalent docking and molecular dynamic (MD) simulations were used to investigate possible avenues of improvement. Rational site-directed mutagenesis of PaL revealed residues V180 and A272 to be the hotspots for diastereopreference. The double V180L/A272F mutant exhibited the highest degree of diastereopreference, as the diastereomeric ratio of (2S, 5R) L-menthol increased towards both (2R, 5S) L-neomenthol (dr1) and (2R, 5R) D-isoneomenthol (dr2) (diastereomeric ratios dr1 and dr2 increased to 4.65 and 2.13 times that of wild-type PaL). MD simulation analysis indicated that these mutations decrease the flexibility of the surrounding protein regions. The combination of increased steric exclusion and decreased flexibility results in less favorable binding of the non-target substrates, (2R, 5S) L-neomenthyl propionate and (2R, 5R) D-isoneomenthyl propionate, to the V180L/A272F mutant. These results confirmed and further improved our previously proposed model of the diastereomer recognition mechanism based on the combined effect of steric exclusion and regional flexibility.     

Genetic and Physical Mapping of the Dreher Locus on Mouse Chromosome 1

Mutations in the mouse dreher (dr) gene cause skeletal defects, hyperactivity, abnormal gait, deafness, white belly spotting, and hypoplasia of Müllerian duct derivatives. To map dr to high resolution, we utilized two crosses. Initially, we analyzed an intersubspecific intercross to construct a detailed genetic map of simple sequence length polymorphism markers within a 6.3-cM region surrounding the dr locus. Subsequently, we analyzed a second intersubspecific intercross segregating for the dr(6J) allele, which positioned dr within a 0.13-cM region between Rxrg and D1Mit370. A physical contig of BAC clones spanning the dr critical region was constructed, and eight potential dr candidate genes were excluded by genetic or physical mapping. Together these results lay the foundation for positional cloning of the dr gene.     

Dynamic kinetic asymmetric transfer hydrogenation of racemic 2,4-diaryl-2,3-dihydrobenzo[b][1,4]diazepines catalyzed by chiral phosphoric acids

Dynamic kinetic Transfer hydrogenation reaction of 2-methyl-2,4-diaryl-2,3-dihydrobenzo[b][1,4]diazepines, using phosphoric acids as catalysts and Hantzsch ester as hydride source, has been studied. A 3,3′-H8-binol derived phosphoric acid has been identified the optimal chiral catalyst for this transformation, affording 1,3-diamine derivatives with up to 8/1 dr, 86% ee and 94% ee for the major and minor diastereomers, respectively.     

I muscoli metamerici delle larve di anuri e la teoria segmentale del Loeb

Il presente lavoro è stato eseguito nell' Istituto di Anatomia e Fisiologia Comparate dell' Università di Palermo, diretto dal prof. F.Raffaele. Il manoscritto relativo era pronto fin dall' ottobre dell' anno passato, ma il mio trasferimento da Palermo a Pavia e varie altre circostanze mi avevano finora impedito dal darvi l'ultima mano.Al prof.Raffaele che ha seguito con efficace interessamento queste ricerche e che ha voluto adesso gentilmente rivedere anche il manoscritto, i miei più cordiali ringraziamenti.     

Design,synthesis and pharmacological evaluation of novel polycyclic heteroarene ethers as PDE10A inhibitors: Part I

We report analogue-based rational design and synthesis of two novel series of polycyclic heteroarenes, pyrrolo[3,2-b]quinolines and pyrido[2,3-b]indoles, tethered to a biaryl system by a methyl-, ethyl- or propyl ether as PDE10A inhibitors. A number of analogues were prepared with variable chain length and evaluated for their ability to block PDE10A enzyme using a radiometric assay. Detailed SAR analyses revealed that compounds with an ethyl ether linker are superior in potency compared to compounds with methyl or propyl ether linkers. These compounds, in general, showed poor metabolic stability in rat and human liver microsomes. The metabolic profile of one of the potent compounds was studied in detail to identify metabolic liabilities of these compounds. Structural modifications were carried out that resulted in improved metabolic stability without significant loss of potency.     

Chaetoglobosins and azaphilones produced by Canadian strains of Chaetomium globosum isolated from the indoor environment

Chaetomium globosum is one of the most common species of fungi found growing on damp building materials in North America and Europe. At doses that could be experienced in a building with some mould damage, exposure to metabolites from other fungi results in inflammatory changes in vivo and in vitro. This research requires knowledge of the dominant toxins produced by fungal strains from the built environment and characterization of pure compounds for toxicity testing. We examined 25 strains of C. globosum isolated from the built environment in Canada. In varying amounts, these strains primarily produced chaetoglobosin A, C and F, chaetomugilin D, and chaetoviridin A. Spectroscopic data of the major isolated compounds are provided. Previous studies reported a number of metabolites from this species that we did not find. However, this appears to be due to misidentifications of the fungi they examined as well as problems with the analytical methods used. In addition, our data support the use of metabolite profiles for resolving the taxonomy of some economically important Chaetomium species.     

The basic flow equations of electrophysiology in the presence of chemical reactions: I. Development of the equations

Starting from the basic flux equation, it is possible to obtain an integral form relating the current componentsI _i at an arbitrary pointr ₂ to the distribution of mobilities and concentrationsc _i, potential forces\(\bar \mu \), and chemical productivityp _i without any restrictive assumptions such as constant mobilities, constant field, steady state, or electrical neutrality. The equation is

$$\begin{gathered} I_i (r_2 ) = G_i (r_2 )\left[ {\Delta \bar \mu _i - \int_{r_1 }^{r_2 } {z_i } FA\left( {p_i - dc_i /dt} \right)\left( {\frac{1}{{G_i (r)}}} \right)dr} \right]; \hfill \\ G_i (r) = 1/\int_{r_1 }^r {\frac{{dr}}{{z_i^2 F^2 c_i u_i }}.} \hfill \\ \end{gathered} $$     

The Deinococcus radiodurans DR1245 Protein,a DdrB Partner Homologous to YbjN Proteins and Reminiscent of Type III Secretion System Chaperones

The bacterium Deinococcus radiodurans exhibits an extreme resistance to ionizing radiation. A small subset of Deinococcus genus-specific genes were shown to be up-regulated upon exposure to ionizing radiation and to play a role in genome reconstitution. These genes include an SSB-like protein called DdrB. Here, we identified a novel protein encoded by the dr1245 gene as an interacting partner of DdrB. A strain devoid of the DR1245 protein is impaired in growth, exhibiting a generation time approximately threefold that of the wild type strain while radioresistance is not affected. We determined the three-dimensional structure of DR1245, revealing a relationship with type III secretion system chaperones and YbjN family proteins. Thus, DR1245 may display some chaperone activity towards DdrB and possibly other substrates.     

Differential display-mediated identification of three drought-responsive expressed sequence tags in tea [Camellia sinensis (L.) O. Kuntze]

There is no information on drought-modulated gene(s) in tea [Camellia sinensis (L.) O. Kuntze], a woody and perennial plant of commercial importance. Using differential display of mRNA, three drought-modulated expressed sequence tags (ESTs) were identified. Northern and BLAST analysis revealed that clonedr1 (droughtresponsive), induced only by drought but not by ABA, showed significant scores with PR-5 (pathogenesis related) family of PR-protein gene. Another clonedr2, repressed by drought but not by ABA, had nucleotide repeats for polyasparate that are also present in chicken calsequestrin-like mRNA. Clonedr3, responded similarly to clonedr2 but did not show significant homology with the reported genes, hence appears to be novel. Identification of these ESTs is an initial step to clone the full length genes and their promoters     

Sullo sviluppo di embrioni completi da blastomeri isolati di uova di tritone (Molge cristata)

Chiudo queste pagine esprimendo al mio illustre maestro prof.Giulio Fano la mia più viva riconoscenza pel modo con cui mi guidÒ in queste ricerche.     

Reliability of utricular function testing using a sinusoidal translation profile during unilateral centrifugation

The unilateral centrifugation test is one of the few vestibular tests that evaluate the utricles side by side. During this test, a subject is rotated about an earth vertical axis at high rotation speeds (e.g. 400 °/s) and translated sideways along the interaural axis to align the axis of rotation consecutively with the right and the left utricle. The combined rotation and translation induces ocular counter rolling (OCR), which is measured using three-dimensional video-oculography. Recently, a new model has been proposed to analyse the OCR. The model is based on contributions from both the semicircular canals and the utricles. Concomitant with the new model a new stimulation profile using a sinusoidal translation profile during the unilateral centrifugation has been introduced [1]. The current study presents the test-retest reliability as well as the robustness of the new stimulation method, based on data of 67 healthy subjects. Test-retest reliability was based on repeated measurements of a group of subjects. To test the robustness of the new sinusoidal translation paradigm, we investigated the effect of a different amplitude of the sinusoidal translation (6 cm instead of 4 cm) and of an offset in translation (from ?3 to +5 cm, instead of from ?4 to +4 cm) on the parameters. Several statistical measures were used to reflect the reliability: intraclass correlation coefficient (ICC), the “coefficient of variation of the method error” and the “minimal difference” (MD). All relevant variables from the physiological model for the OCR induced by unilateral centrifugation show a good to excellent reliability during the test-retest study and the relevant parameters remain unaffected by the changes applied to the translation profile (p > 0.05) as predicted by the model. Additionally, all observed differences are smaller than the MD values calculated in the test-retest part of the study.