Hyperferritinemia -cataract syndrome: Long-term ophthalmic observations in an Italian family

Background Hyperferritinemia-cataract syndrome (HCS) is a rare Mendelian condition characterized by bilateral cataract and high levels of serum ferritin in the absence of iron overload. Methods HCS was diagnosed in three adult siblings. In two of them it was possible to assess lens changes initially in 1995 and again in 2013. Serum ferritin, iron, transferrin concentrations and transferrin saturation percentage were also measured, and the Iron Responsive Element (IRE) region of the L-ferritin gene (FTL) was studied. Results Serum ferritin concentrations were considerably elevated while serum iron, transferrin and transferrin saturation levels were within the normal range in each sibling. Cataract changes in our patients were consistent with those previously reported in the literature. Progression of the cataract, an aspect of few studies in this syndrome, appeared to be quite limited in extent. The heterozygous +32G to T (-168G>T) substitution in the IRE of the FTL gene was detected in this family. Conclusions Ophthalmic and biochemical studies together with genetic testing confirmed HCS in three family members. Although the disorder has been extensively described in recent years, little is known regarding cataract evolution over time. In our cases, lens evaluations encompassed many years, identified bilateral cataract of typical morphology and supported the hypothesis that this unique clinical feature of the disease tends to be slowly progressive in nature, at least in adults.

Sequence of reactions leading to cytochrome-p-450 synthesis-effect of drugs

The effect of phenobarbital on the rates of the synthesis of the protein and heme moieties of cytochrome P-450 has been studied. For this purpose, cytochrome P-450 has been partially purified as its P-420 derivative and the labeled amino acid incorporation into the protein has been studied after subjecting a partially purified preparation to sodium dodecyl sulfate gel electrophoresis. The incorporation studies into the protein species after sodium dodecyl sulfate gel electrophoresis reveal that the drug primarily accelerates the rate of apoprotein synthesis followed by an increase in the rate of heme synthesis. The messenger for apocytochrome P-450 appears to be fairly stable.

Italian ja Veneton l����nin kolmas sektori. Maaraportti maaseutualueiden palvelutuotannon haasteista ja hyvist�� k��yt��nn��ist��.

Tiivistelm�� Maaraportin tavoitteena on luoda yleiskatsaus Italian kolmanteen sektoriin ja tutkimus kohdistuu erityisesti Pohjois-Italiassa sijaitsevan Veneton l����nin kolmannen sektorin palvelutuotantoon. Raportissa tarkastellaan erityisesti kolmannen sektorin ja julkisen palvelutuotantosektorin sek�� julkishallinnon toiminnallisia suhteita. Kolmannen sektorin mahdollisuuksia vastata maaseutualueiden palvelujen tarjonnan haasteisiin on pyritty analysoimaan. Italia on julkishallinnon ja kolmannen sektorin palvelutuotannon kannalta tarkasteltuna nykytilanteessa mielenkiintoinen testilaboratorio koko Euroopan mittakaavassa johtuen maan historiallisista, kulttuuri-sista, kieli- ja maantieteellisist�� sek�� itse v��est��pohjan tarjoamista haasteista. Italiasta ei voida puhua yhden homogeenisen valtiok��sitteen alla, koska maa toimii kulttuurisista perinteist����n johtuen kolmella eri nopeudella. Itse valtio k��sitett��kin tulkitaan Italiassa monin eri tavoin pohjautuen ���valtiona��� n��hdyn organisaation k��yt��nn��n funktioon sitoutuneena alueellisiin ja kulttuurisiin tekij��ihin. Teollinen, dynaaminen yritysten varakas Pohjois-Italia henkil��ityneen�� Milanoon, hallinnon ja kulttuurin leimaama Keski-Italia kiteytyneen�� ikuiseen kaupunkiin Roomaan ja Napolin alapuolinen Etel��-Italia, joka painii ���j��rjestelm��ss�� sis��ll�� olevan j��rjestelm��ns����� kanssa eiv��t ole koskaan muodos-taneet oikeasti yhten��ist�� Italian valtiota. Italiassa erillisin�� itsehallinnollisina alueina toimivat l����neiksi luettavat Sisilia, Trentino Alto-Adige, Val di Aosta, Sardegna ja Friuli-Venezia Giulia, mik�� lis���� hallinnollista kirjavuutta. Viimeisten vuosien aikana Italian julkishallinto on ollut jatkuvassa transitiotilassa yritt��ess����n kohdata uudistumisen vaateita globalisoituvassa maailmassa. Italiassa on meneill����n hidas siirtyminen liittovaltiomaiseen j��rjestelm����n ja jopa syvemp����n paikallisuuteen. Massiivinen ja raskas julkishallintokoneisto nelj��ll�� eri tasolla (valtio - l����ninhallinnolliset alueet - vahvat maakunnat - kunnat) on eritt��in hankalasti modernisoitavissa, mutta poliittinen tahtotila on vaihtumassa klassista vahvaa keskushallintoa kannattavasta ajattelutavasta hyv��ksym����n federalistisia, alueellisia hallintoratkaisuja. Italiassa on her��tty havaitsemaan, ett�� ongelmien ratkaisu t��ytyy vied�� itse alueille ja niiden ihmisille, koska unilateraalinen keskushallinnointitapa ei en���� vastaa monikerroksisiin, moniulotteisiin alueellisiin haasteisiin. Jokaisella alueella on erilaiset l��ht��kohdat ongelmien ratkaisuihin my��s globalisoitumisen asettamissa yhteisiss�� haasteissa. Kolmannen sektorin asema arvostettuna italialaisten kansalaispalvelujen t��ydent��j��n�� ja tuottajana alkaa olla nykyp��iv����. Kolmannen sektorin elint��rke�� osuus uusien palvelutarpeiden tunnistamisessa on jo yleisesti tunnustettu tosiasia Italiassa. Vapaaehtoisj��rjest��t toimivat eturiviss�� kansalaisten arkip��iv��ss�� ja pystyv��t n��in kanavoimaan ensimm��isin�� sosiaalista kehityskulkua ja kohottamaan esiin erilaisia marginaalistenkin ryhmien tarpeita sek�� vastaamaan niihin. Kolmas sektori laajentaa, tuottaa ja paikkaa paikallista julkista palvelutuotantoa. Ratkaiseva askel lains����d��nn��lliselt�� kannalta on ollut rahoitusvirtojen ohjaaminen kolmannen sektorin ns. ONLUS-organisaatioille ja ONLUS-statuksen perustaminen vuonna 1997. Italiassa on 60 231.214 asukasta (31.7.2009) ja se on hallinnollisesti jakaantuneena 8 100 kuntaan. Suomessa vastaavat luvut ovat 5 350 712 asukasta ja 348 kuntaa (11/2009). Maantieteelliselt�� kooltaan Italia ei eroa 301 338 km2:n pinta-alallaan paljonkaan Suomesta (338 424��km2). V��est��m����r��n erojen vuoksi Italiassa on keskim����rin 199,9 asukasta neli��kilometrill�� ja Suomessa 17,1. Maiden v��linen suora vertailu ei ollut mittakaavaerojen vuoksi j��rkev����. Kuntien hallinnollinen rooli on Italiassa painotukseltaan erilainen kuin Suomessa. Suomessa kuntien vastuulla oleva sosiaali-, terveydenhuolto- ja koulutoimi eiv��t ole samassa laajuudessa italialaisten kuntien vastuulla vaan n��iden toimialojen vastuulliset tahot ovat l����nit terveydenhoitopiireineen sek�� maakuntahallinto koulupiirien osalta. Italialaisten kuntien vastuualueet voivat my��s vaihdella mittavasti perustuslaissa m����riteltyjen perustoimien lis��ksi. Tutkimuksen kohteeksi on rajattu Koillis-Italiassa sijaitseva Veneton l����nin alue, koska se on v��kim����r��lt����n l��hes yht�� suuri kuin Suomi, 4 893��309 asukasta (31.3.2009). Kuntien lukum����r�� on huimaava 581 kpl. Veneto on maantieteelliselt�� alueeltaan pieni ja harvaan asuttua maaseutua on pinta-alasta melko niukasti. Pohjoisen osan vuoristoalueet karuine olosuhteineen muodostavat palvelutuotannollisesti haas-teellisen ymp��rist��n, joka on verrattavissa suomalaisten maaseutualueiden tilanteeseen. Venetossa on omaksuttu Italian mittakaavassa innovatiivisia ratkaisuja julkishallinnon palvelutuotannon ja kolmannen sektorin toiminnan osalta. Veneton alue on luokiteltu maailman pienyritysintensiivisimm��ksi alueeksi, mik�� heijastuu my��s alueen palvelutuotantoratkaisuissa. Raportti esittelee ensin Italian hallintoa, keskushallinnon, paikallishallinnon ja tutkimuksen asettelulle olennaisten kolmannen sektorin toimijoiden osalta. T��m�� on v��ltt��m��t��nt��, sill�� kolmannen sektorin toiminta on Italiassa hyvin pitk��lle lains����d��nn��ll�� ohjattua ja rajattua. Kolmannen sektorin k��sitteist����, toimijoita, j��rjest��ytymist�� ja sen toimintaa s����telev���� lains����d��nt���� esitell����n tarkemmin. Veneton l����nin osalta kolmanteen sektoriin perehdyt����n sek�� hallinnon ett�� toimijakent��n n��k��kulmia tulkiten. Raportin lopussa esitell����n kolmannen sektorin palvelutuotantoon ja hallinnointiin liittyvi�� case-esimerkkitapauksia.

Biotransformations of ��-terpineol in the rat: Its effects on the liver microsomal cytochrome P-450 system

Alpha-Terpineol (I), a monocyclic monoterpene tertiary alcohol, is widely used in the manufacture of perfumes, cosmetics, soaps and antiseptic agents. It was reported earlier (Horning et al. 1976) that this monoterpene alcohol when administered to humans is hydroxylated to p-menth-l,2,8-triol (II). It is not known whether c~-terpineol also produces other metabolites during its metabolism in the mammalian system and if so, the nature of these metabolites.

Heme is a positive regulator of cytochrome P-450 gene transcription

Inhibitors of heme biosynthesis such as CoCl2, 3-amino-1,2,4-triazole, and thioacetamide block the 3-methylcholanthrene-mediated induction of cytochrome P-450 (c + d) messenger RNAs and their transcription in rat liver. This effect is specific, since the messenger RNA levels for albumin and glutathione transferase (Ya + Yc) and their transcription are not significantly influenced under conditions of heme depletion. Exogenous administration of heme at very low doses (50 ��g/100 g body wt) is able to completely counteract the effects of the heme biosynthetic inhibitors on cytochrome P-450 (c + d) messenger RNA levels and their transcription. This constitutes a direct proof for the role of heme as a positive regulator of cytochrome P-450 gene transcription.

Heme regulates cytochrome P-450 gene transcription elongation

Administration of 3-methylcholanthrene (MC) to rats results in a striking increase in the transcription of cytochrome P-450 (c+d) messenger RNA with isolated nuclei, which is blocked by the simultaneous administration of cobalt chloride, an inhibitor of heme biosynthesis. Transcription of cytochrome P-450 (c+d) mRNAs with nuclei isolated from MC treated rats shows a linear increase with time of incubation, whereas it shows a progressive decrease with incubation time in the case of nuclei isolated from MC+CoCl2 treated rats. Addition of heme in vitro (10���6M) to the latter nuclei results in a significant counteraction of the decreased cytochrome P-450 (c+d) mRNA transcription. The inhibition in transcription rates observed in MC+CoCl2 treated rat liver nuclei is more pronounced with the seventh exon probe than with the second exon probe. Once again, in vitro heme addition can counteract the inhibition observed with both the probes. Since run off transcription with isolated nuclei represents essentially elongation of the initiated transcripts, the data obtained can be interpreted on the basis that heme regulates cytochrome P-450 gene transcription elongation.

Site of synthesis of cytochrome P-450 in liver

The suggestion that a rapidly sedimenting rough endoplasmic reticulum fraction in close association with mitochondria, is the preferred site of cytochrome P-450 synthesis has been examined. The rate of cytochrome P-450 synthesis in the different subcellular fractions has been evaluated Image , using the immunoprecipitation technique. The results indicate that the conventional microsomal fraction (100,000 X g sediment) is the major site of cytochrome P-450 synthesis and that the rapidly sedimenting rough endoplasmic reticulum fraction associated with mitochondria is not a preferred site for the hemoprotein synthesis.

Destruction Of Rat-Liver Microsomal Cytochrome-P-450 Invitro By A Monoterpene Ketone, Pulegone A Hepatotoxin

Significant destruction (68%) of liver microsomal cytochrome P-450 and homogeneous cytochrome P-450 purified from PB-treated rats is noticed upon incubation with 10 mM pulegone at 37-degrees-C for 30 min. There is also a concomitant loss of heme. The destructive phenomenon does not require metabolic activation of pulegone. The destruction of purified cytochrome P-450 is time-dependent and saturable. Structure-activity studies suggest that an alpha-isopropylidine ketone unit with a methyl positioned para to the isopropylidine group as in pulegone is necessary for the in vitro destruction of cytochrome P-450. SKF-525A at a concentration of 4-mM obliterates the destruction of cytochrome P-450 by pulegone. Experiments with C-14-pulegone suggest that pulegone or its rearranged product binds covalently to the prosthetic heme of cytochrome P-450.

Estradiol - 17�� induces polyaromatic hydrocarbon-inducible cytochrome P-450 in chicken liver

A cDNA clone has been isolated from a chicken liver library prepared against messenger RNA isolated after chronic estradiol-17�� treatment. The clone, pP-450 IA - 61, has an insert of 900nt and the sequence shows high homology to CYPIA2 subfamily from four other species. A single injection of estradiol-17�� to immature chicken results in a striking induction of mRNA hybridizing to labeled pP-450IA - 61. The probe also hybridizes to mRNA induced by 3 ��� methylcholanthrene in chicken. These results offer direct proof for the similarity in the mode of action at the transcriptional level of polyaromatic hydrocarbons and estrogenic compounds.

Studies on the biosynthesis of cytochrome P-450 in rat liver���A probe with phenobarbital

Cytochrome P-450 has been purified from phenobarbital-treated rat livers to a specific content of 17 nmol/mg protein. The major species purified has a molecular weight of 48,000. Using the purified antibody for the cytochrome P-450 preparation it has been shown that the major product synthesized in vivo and in the homologous cell-free system in vitro is the 48,000 molecular weight species. Poly(A)-containing RNA isolated from phenobarbital-treated animals codes for the synthesis of the 48,000 molecular weight species in the wheat germ and reticulocyte lysate cell-free systems. It is concluded that cytochrome P-450 synthesis does not involve processing of a polyprotein precursor, although certain minor modifications including glycosylation of the primary translation product are not ruled out. Phenobarbital treatment of the animal results in a significant increase in the cytochrome P-450 messenger activity as measured in the wheat germ cell-free system.

Microsomal redox systems in brown adipose tissue: high lipid peroxidation, low cholesterol biosynthesis and no detectable cytochrome P-450

The presence of redox systems in microsomes of brown adipose tissue (BAT) in cold exposed rats was investigated and compared with liver. BAT microsomes showed high activity of lipid peroxidation measured both by the formation of malondialdehyde (MDA) and by oxygen uptake. NADH and NADPH dependent cytochrome c reductase activity were present in both BAT and liver microsomes. Aminopyrine demethylase and aniline hydroxylase activities, the characteristic detoxification enzymes in liver microsomes could not be detected in BAT microsomes. BAT minces showed very poor incorporation of [1-14C]acetate and [2-14C]-mevalonate in unsaponifiable lipid fraction compared to liver. Biosynthesis of cholesterol and ubiquinone, but not fatty acids, and the activity of 3-hydroxy-3-methyl glutaryl CoA reductase appear to be very low in BAT. Examination of difference spectra showed the presence of only cytochrome b 5 in BAT microsomes. In addition to the inability to detect the enzyme activities dependent on cytochrome P-450, a protein with the characteristic spectrum, molecular size in SDS-PAGE and interaction with antibodies in double diffusion test, also could not be detected in BAT microsomes. The high activity of lipid peroxidation in microsomes, being associated with large oxygen uptake and oxidation of NADPH, will also contribute to the energy dissipation as heat in BAT, considered important in thermogenesis.