Search PubMed⌕ Search

Biomedical subjects

M Huber

Publications and source records attributed to M Huber.

At least 163 records · Page 9Linked to original sources
PubMed1994

Crystallographic analyses of site-directed mutants of the photosynthetic reaction center from Rhodobacter sphaeroides.

Seven site-directed mutants of the bacterial photosynthetic reaction center (RC) from the 2.4.1 and WS 231 wild-type strains of Rhodobacter sphaeroides have been crystallized and their X-ray diffraction analyzed to resolutions between 3.0 and 4.0 A. The mutations can be divided into four distinct categories: (1) mutations altering cofactor composition that affect electron transfer and quantum yield, His M202-->Leu (M202HL), His L173-->Leu (L173HL), and Leu M214-->His (M214LH); (2) a mutation in the proposed pathway of electron transfer altering electron-transfer kinetics, Tyr M210-->Phe (M210YF); (3) a mutation around the non-heme iron resulting in an iron-less reaction center, His M219-->Cys (M219HC); and (4) mutations around the secondary electron acceptor, a ubiquinone, affecting proton transfer and quinone turnover, Glu L212-->Gln (L212EQ) and Asp L213-->Asn (L213DN). Residues L173 and M202 are within bonding distance of the respective magnesiums of the two bacteriochlorophylls of the BChl special pair, while M214 is close to the bacteriopheophytin on the active A branch of the RC. The L173HL and M202HL crystal structures show that the respective bacteriochlorophylls are replaced with bacteriopheophytins (i.e., loss of magnesium) without significant structural perturbations to the surrounding main-chain or side-chain atoms. In the M214LH mutant, the bacteriopheophytin has been replaced by a bacteriochlorophyll, and the side chain of His M214 is within ligand distance of the magnesium. The M210YF, L212EQ, and L213DN mutants show no significant tertiary structure changes near the mutation sites. The M219HC diffraction data indicate that the overall tertiary structure of the reaction center is maintained in the absence of the non-heme iron.

Chemical Phenomena↗
PubMed1994

Mutations in the rod domain of keratin 2e in patients with ichthyosis bullosa of Siemens.

Ichthyosis bullosa of Siemens (IBS) is an autosomal dominant skin disorder that resembles epidermolytic hyperkeratosis (EHK). We have identified mutations in two families originally diagnosed with EHK and in four families diagnosed with IBS at the same codon in the highly conserved carboxy terminal of the rod domain of keratin 2e, thus revealing a mutational hot spot. Our results allow a differential diagnosis to be made between IBS and EHK at the genetic level and we suggest that patients diagnosed with EHK, but lacking keratin K1 or K10 mutations, should be re-examined for mutations in their K2e genes.

Adult↗
PubMed1994

Primary therapy of multiple myeloma with paclitaxel (taxol).

BACKGROUND: The treatment of multiple myeloma remains unsatisfactory and new active agents are needed. Paclitaxel is effective against a variety of solid tumors and we assessed the utility against multiple myeloma. PATIENTS AND METHODS: From March 1993 to May 1994, we treated 33 patients with newly diagnosed multiple myeloma with paclitaxel given intravenously at a dose of 125 mg/m2 over 24 hours (13 patients) or at a dose of 135 mg/m2 over 3 hours (20 patients). RESULTS: Five of 33 patients responded (15%; 95% CI: 5 to 32%) with an unmaintained remission of 3-11+ months. Severe but reversible neutropenia was the major dose limiting toxicity, but myalgias and alopecia were also common. CONCLUSION: Paclitaxel was slightly active against multiple myeloma. Whether higher doses or new analogues of this agent can produce superior results requires further study.

Adult↗
PubMed1994

Abnormal keratin 1 and 10 cytoskeleton in cultured keratinocytes from epidermolytic hyperkeratosis caused by keratin 10 mutations.

Epidermolytic hyperkeratosis is caused by mutations of the differentiation-specific keratins K1 and K10. These mutations produce a weakened cytoskeleton that is prone to collapse resulting in cell fragility and lysis. In this study we have analyzed cultured keratinocytes from EHK patients bearing 10R-to-H and 15L-to-S mutations within the 1A segment of the K10 rod domain. Keratinocytes were grown submerged in serum-free medium and induced to differentiate by growing to confluence and increasing the Ca++ concentration in the medium. Cultures were either harvested for mRNA sequence analysis or subjected to immunofluorescence microscopy. Differentiating keratinocytes from these patients were found to express these K10 mutations in their mRNA. Moreover, these cells could be distinguished from normal keratinocytes by their aberrant morphology. EHK keratinocytes frequently exhibited a collapsed perinuclear network of K1/K10 filaments and sometimes peripheral granules of K1 and K10 aggregates, reminiscent of the cells of the suprabasal layers in these patients. This report documents the expression of mutant keratin 10 in cultured EHK keratinocytes.

Base Sequence↗
PubMed1994

Involucrin mRNA is more abundant in human hair follicles than in normal epidermis.

Involucrin is a precursor protein of the cornified cell envelope in epidermal keratinocytes, where it has been located by immunohistochemistry in the upper spinous and granular layers of human epidermis. In the hair follicle, involucrin has been found in the inner root sheath and in the upper layers of the infundibulum and the isthmus (upper outer root sheath), whereas its presence in the lower outer root sheath and the cortex has been controversial. Therefore, we analyzed the distribution of involucrin mRNA in adult scalp by Northern blotting and in situ hybridization. Northern blots showed more abundant involucrin mRNA in the follicular fraction than in the epidermal fraction of dissected scalp. In situ hybridization matched the immunohistologic results; transcripts of involucrin were expressed not only in the infundibulum and isthmus, but also in the hair cortex and medulla, in all layers of the inner root sheath, and in the inner cells of the lower outer root sheath (all of which lack a cell envelope at the ultrastructural level). However, involucrin was absent in the hair cuticle, which is the only compartment of the follicle possessing a morphologically distinct cell envelope. Our results suggest, first, that involucrin does not serve as a precursor protein of the cornified cell envelope in adult hair follicles, and second, that it is perhaps not necessary for the formation of the cell envelope in keratinocytes of the hair cuticle, as we did not find this precursor protein with highly sensitive methodology.

Blotting, Northern↗
PubMed1994

[Resection of cervical glands in pelviscopic hysterectomy].

A case of pelviscopic hysterectomy with uterine hyperflexion and cervical elongation is presented. In this case we were forced to extirpate the cervical fascia because of uterine hemorrhage by laparotomy after the cervical tissue punch had been performed. The histological findings were supravaginal cervical glands. Complete removal of cervical glandular tissue may be undetermined by hyperflected uteri and cervical elongation.

Adult↗
PubMed1994

Histology of tissue adjacent to an HAC-coated femoral prosthesis. A case report.

We studied the fixation of a cementless titanium femoral prosthesis partially coated with hydroxyapatite ceramic (HAC) 10.4 months after implantation. Histomorphological investigation showed extensive new bone formation between the HAC coating and the bone bed; morphometry showed bone contact indices of up to 91.60%. There were a number of resorption lacunae on the HAC coat with depths of up to 76.6 microns and widths of up to 453 microns. Our results confirmed that considerable bone remodelling had taken place and that the apatite-coated prosthesis had united with bone despite the lack of appreciable immediate press-fit. Hydroxyapatite particles which had been released did not appear to show any negative effects on the stability of the implant.

Aged↗
PubMed1993

The mannose transporter of Escherichia coli. Structure and function of the IIABMan subunit.

UNLABELLED: The mannose transporter of the bacterial phosphotransferase system consists of two transmembrane subunits (IICMan and IIDMan) and a hydrophilic subunit (IIABMan). IIABMan has two flexibly linked domains containing one phosphorylation site each and occurs as a dimer. Substrate transport is coupled to phosphorylation. The phosphoryl group is transferred from a phosphoryl carrier protein to His10 on IIA, hence to His175 on IIB and finally to the substrate. IIABMan mutants were analyzed in vitro for complementation, negative dominance, cysteine cross-linking and reactivity. CONCLUSIONS: (i) His10, Trp12, Lys48, and Ser72 form a functional unit (phosphorylation site 1); (ii) His86 on the IIA domain and His175 on the IIB domain of the same subunit form a functional unit (phosphorylation site 2); (iii) phosphoryl transfer can occur between His10 and His175 of the same as well as of different subunits and His86 is necessary for this transfer; (iv) the subunits in the dimer are interdependent; (v) The phosphorylation site mutant H175C is highly reactive toward thiol reagents and it forms extensive homo- and heterocross-links with other surface-exposed cysteines. The phosphorylation site mutant H10C is 1000-fold less reactive. The two residues might be in complementary locations, His10 buried in a concave, His175 exposed on a convex surface.

Bacterial Proteins↗
PubMed1993

Pontine infarction in acute posterior multifocal placoid pigment epitheliopathy.

In a patient with acute posterior multifocal placoid pigment epitheliopathy (APMPPE), a pontine infarction occurred about 6 months after the ophthalmological manifestation. We report the first case with histopathologically proven vasculitis shown by muscle biopsy and the first positron emission tomographic documentation in APMPPE. The ophthalmological and cerebral symptoms responded well to steroid treatment. Long-term immunosuppression (e.g. azathioprine 1-2 mg/kg) seems to decrease the risk of recurrent systemic vasculitis.

Acute Disease↗
PubMed1993

Physical dependence on nicotine gum: effect of duration of use.

This study examined whether longer duration on nicotine gum promoted dependence on nicotine gum. Subjects (N = 128) answering an advertisement for smoking cessation research and wanting to quit smoking cigarettes were randomly assigned to 1- or 3-month duration of nicotine gum use. Assessments were made weekly for smoking status (with biochemical verification) and withdrawal symptoms during and at the end of treatment. Follow-up was conducted at 1, 6 and 12 months to provide exploratory data on treatment outcome. The results showed minimal nicotine gum withdrawal symptoms after gum cessation with virtually no difference in gum withdrawal between the 1- and 3-month groups. Withdrawal symptoms from the nicotine gum included difficulty with concentration, increased variability on a reaction time task, and decreased vigor. The results also showed that continuous use of the gum at 1 year was observed in 1.5% of subjects and estimated to be as high as 14%. Finally, the 3-month group experienced a 2-fold increase in abstinence compared to the 1-month group, although this difference was not statistically significant. We conclude that there is minimal physical dependence on nicotine gum.

Adult↗
PubMed1993

Effect of propentofylline on regional cerebral glucose metabolism in acute ischemic stroke.

In a randomized double-blind placebo-controlled study in 30 patients with acute ischemic stroke, the effect of the adenosine uptake blocker propentofylline on regional brain glucose metabolism (rCMRglu) was investigated using repeated positron emission tomography (PET) with 2-[18F]fluoro-2-deoxy-D-glucose (FDG). Treatment was initiated within 48 h after onset of symptoms. The clinical course was followed for 3 months. In the propentofylline group, after 14 days rCMRglu was increased in the infarct by 37.3% and was practically unchanged in other brain regions, whereas in the control group glucose metabolism had decreased in all regions (1.4-13.4%). These differences were significant between the two groups [Analysis of variance (ANOVA) p = 0.005]. Although there was a trend toward greater clinical improvement in the propentofylline-treated patients, this did not reach statistical significance. The results correspond to experimental data showing that propentofylline improves energy metabolism in cerebral ischemia. A clinical trial is needed to determine whether this new therapeutic principle can be successfully used in acute human stroke.

Acute Disease↗
PubMed1993

A mutational hot spot in keratin 10 (KRT 10) in patients with epidermolytic hyperkeratosis.

Epidermolytic hyperkeratosis (EHK), (bullous congenital ichthyosiform erythroderma), is an autosomal dominant human skin disorder. Recently, we and others have described mutations in keratins 1 and 10 (K1 and K10) in patients with this disease. Structure-function models predict that these mutations would impair normal filament assembly and function. We have extended our earlier studies to include 8 more incidences of EHK. In half of these families, we were unable to locate a mutation within the rod domains of either K1 or K10. However, polymorphic restriction site and sequence analysis of the other families revealed a mutational hot spot within the 1A alpha-helical segment of K10. These involve Arginine to Histidine, Arginine to Cysteine and Arginine to Leucine substitutions at residue 10 of the rod domain. Interestingly, mutations in the corresponding Arginine residue in keratin K14 have been identified in patients with epidermolysis bullosa simplex. The large number of mutations found at this position in both keratins K10 and K14 suggests that other epithelia cell disorders will be discovered that are caused by the corresponding mutation in related type I keratin genes.

Amino Acid Sequence↗
PubMed1993

Expression patterns of loricrin in various species and tissues.

In this study we analyzed the expression patterns of loricrin in various species and tissues using immunohistochemistry, immunoblotting and Northern blots. Loricrin is a glycine-, serine- and cysteine-rich protein expressed very late in epidermal differentiation in the granular layers of normal mouse and human epidermis. Later on in differentiation, loricrin becomes crosslinked as a major component into the cornified cell envelope by the formation of N epsilon-(gamma-glutamyl)lysine isopeptide bonds. This process either occurs directly or by the intermediate accumulation in L-keratohyaline granules of mouse epidermis and human acrosyringia. Loricrin was identified in all mammalian species analyzed by virtue of its highly conserved carboxy-terminal sequences revealing an electric mobility of approximately 60 kDa in rodents, rabbit and cow and of approximately 35 kDa in lamb and human on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Loricrin is expressed in the granular layer of all mammalian orthokeratinizing epithelia tested including oral, esophageal and fore-stomach mucosa of rodents, tracheal squamous metaplasia of vitamin A deficient hamster and estrogen induced squamous vaginal epithelium of ovary ectomized rats. Loricrin is also expressed in a few parakeratinizing epithelia such as BBN [N-butyl-N-(4-hydroxybutyl)nitrosamine]-induced murine bladder carcinoma and a restricted subset of oral and single vaginal epithelial cells in higher mammals. Our results provide further evidence that the program of squamous differentiation in internal epithelia of the upper alimentary tract in rodents and higher mammals differ remarkably. In addition, we also have noted the distinct distribution patterns of human loricrin and involucrin, another major precursor protein of the cornified cell envelope.

Animals↗