Human mitochondrial complex I deficiency: investigating transcriptional responses by microarray.

NADH:ubiquinone oxidoreductase (complex I) deficiency is one of the most frequently encountered defects of the mitochondrial energy generating system. A deficiency of this enzyme complex leads to a wide variety in clinical disease expression. The cell biological consequences of such mutations, however, are poorly understood. We investigated transcriptional responses in fibroblast cell lines harboring mutations in the five different nuclear DNA encoded subunits using a mitochondria-targeting microarray. Expression profiles of cell lines cultured under conditions that favor glycolytic metabolism were compared to profiles when cultured under conditions favoring oxidative metabolism. Approximately 60 genes displayed differential expression under these conditions in either all mutated cell lines or selected cell lines only. A marked induction of metallothioneins as well as ATP1G1 transcripts was detected in all patient cell lines. Transcriptional responses such as the induction of heat shock protein transcripts, decreased PDK1,BNIP3 and mitochondrial genome encoding gene transcripts occurred in selected patient cell lines. The observed transcript profile points to a common, putative defensive, response relating to oxidative stress. Although further investigations of other human OXPHOS system diseases is warranted, these results clearly underline that functional genomics holds for the study of inherited metabolic disease.

The mitochondrial genome of the sperm whale and a new molecular reference for estimating eutherian divergence dates.

Extant cetaceans are systematically divided into two suborders: Mysticeti (baleen whales) and Odontoceti (toothed whales). In this study, we have sequenced the complete mitochondrial (mt) genome of an odontocete, the sperm whale (Physeter macrocephalus), and included it in phylogenetic analyses together with the previously sequenced complete mtDNAs of two mysticetes (the fin and blue whales) and a number of other mammals, including five artiodactyls (the hippopotamus, cow, sheep, alpaca, and pig). The most strongly supported cetartiodactyl relationship was: outgroup,((pig, alpaca), ((cow, sheep),(hippopotamus,(sperm whale,(baleen whales))))). As in previous analyses of complete mtDNAs, the sister-group relationship between the hippopotamus and the whales received strong support, making both Artiodactyla and Suiformes (pigs, peccaries, and hippopotamuses) paraphyletic. In addition, the analyses identified a sister-group relationship between Suina (the pig) and Tylopoda (the alpaca), although this relationship was not strongly supported. The paleontological records of both mysticetes and odontocetes extend into the Oligocene, suggesting that the mysticete and odontocete lineages diverged 32-34 million years before present (MYBP). Use of this divergence date and the complete mtDNAs of the sperm whale and the two baleen whales allowed the establishment of a new molecular reference, O/M-33, for dating other eutherian divergences. There was a general consistency between O/M-33 and the two previously established eutherian references, A/C-60 and E/R-50. Cetacean (whale) origin, i.e., the divergence between the hippopotamus and the cetaceans, was dated to approximately 55 MYBP, while basal artiodactyl divergences were dated to >/=65 MYBP. Molecular estimates of Tertiary eutherian divergences were consistent with the fossil record.

Strain, clone and species: comments on three basic concepts of bacteriology.

Different aspects of the terms strain, clone and species are discussed. The term strain is commonly used to denote a pure culture - here called 'the strain in the taxonomic sense' - but does also refer to a natural concept closely related to the clone. The term clone on the other hand is used both in a general and in a more restricted sense, the latter indicating a low degree of genetic exchange. The important distinction between the definition of a species and the criteria for a species is emphasised and the main kinds of criteria are considered.

The complete mitochondrial DNA sequence of the pig (Sus scrofa).

The complete mitochondrial genome sequence of the pig, Sus scrofa, was determined. The length of the sequence presented is 16,679 nucleotides. This figure is not absolute, however, due to pronounced heteroplasmy caused by variable numbers of the motif GTACACGTGC in the control region of different molecules. A phylogenetic study was performed on the concatenated amino acid and nucleotide sequences of 12 protein-coding genes of the mitochondrial genome. The analysis identified the pig (Suiformes) as a sister group of a cow/whale clade, making Artiodactyla paraphyletic. The split between pig and cow/whale was molecularly dated at 65 million years before present.

Phylogeny of nematode-trapping fungi based on 18S rDNA sequences.

The small subunit (SSU) ribosomal DNA (18S rDNA) from 15 species of nematode-trapping fungi and closely related non-parasitic species were sequenced. Phylogenetic analysis indicated that species within the genera of Arthrobotrys, Dactylaria, Dactylella, Monacrosporium and Duddingtonia formed a monophyletic and isolated clade among an unresolved cluster of apothecial ascomycetes. The phylogenetic patterns within this clade were not concordant with the morphology of the conidia nor the conidiophores, but rather with that of the infection structures. The results from the different methods of tree reconstruction supported three lineages; the species having constricting rings, the non-parasitic species and the species having various adhesive structures (nets, hyphae, knobs and non-constricting rings) to infect nematodes.

Analyses of mitochondrial genomes strongly support a hippopotamus-whale clade.

Although the sister-group relationship between Cetacea and Artiodactyla is widely accepted, the actual artiodactyl group which is closest to Cetacea has not been conclusively identified. In the present study, we have sequenced the complete mitochondrial genome of the hippopotamus, Hippopotamus amphibius, and included it in phylogenetic analyses together with 15 other placental mammals. These analyses separated the hippopotamus from the other suiform included, the pig, and identified the hippopotamus as the artiodactyl sister group of the cetaceans, thereby making both. Artiodactyla and the suborder. Suiformes paraphyletic. The divergence between the hippopotamid and cetacean lineages was calculated using this molecular data and was estimated at ca. 54 Ma BP.

Psychological profile of patients suffering from Crohn's disease and ulcerative colitis.

Patients with Crohn's disease (CD, n = 61) and ulcerative colitis (UC, n = 24) were studied using an interview and a series of percept-genetic tests describing anxiety, defensive strategies, and the self-image. Attention was specifically directed at a subgroup of CD (n = 15) with its inflammation limited to the colon (colon Crohn, CC). There was also a control group (n = 43). CC had more immature parent relations than the other groups. They were more childishly concrete and more alexithymic. While CC differ from other Crohn patients in many respects, the real cleft opens up with regard to UC, in spite of location of their illness. CC patients obviously have a psychological profile of their own.

Septicemia in granulocytopenic patients: a shift in bacterial etiology.

35 episodes of septicemia in 33 patients occurred among 269 consecutive patients with granulocytopenia (granulocyte cell count less than or equal to 0.5 x 10(9)/l) during the 7-year period 1982-1988. 59% of isolated bacteria were Gram-positive (Staphylococcus aureus, Staph. epidermidis, Streptococcus species and Pneumococcus) and 41% Gram-negative (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Enterobacter, Acinetobacter and Citrobacter). Compared to the 7-year period 1975-1981, there was a decrease in the relative number of patients with Gram-negative septicemia. Thus, a shift from a predominating Gram-negative etiology in the 1975-1981 period to a predominating Gram-positive etiology in the 1982-1988 period was noted. In both periods the mortality rate was high in patients with Gram-negative septicemia, especially in patients with a Ps, aeruginosa infection.

Neurophysiologic studies of patients with Crohn's disease on long-term treatment with metronidazole.

Fifty-three patients with Crohn's disease were chosen for this study. They were divided into three groups: the first group (19 patients) was treated with metronidazole and had been receiving the drug for at least 1 year; the second group (13 patients) had previously had the same type of long-term treatment but had been off the drug for at least 3 months before entering the study; the third group (21 patients) had never received treatment with metronidazole. The maximum daily dose of metronidazole used was 800 mg. All patients were examined with a broad set of neurophysiologic investigations devised to reveal signs of peripheral neuropathy. No significant difference was found between the three groups. The conclusion was that a daily dose of metronidazole not exceeding 800 mg is well tolerated by patients with Crohn's disease with regard to objective signs of neurotoxicity. A few patients from all three groups showed a raised perception threshold for changes of temperature in the feet (as compared with the department's normal standard). The results of the test were almost identical in the three groups and thus not connected with the metronidazole treatment. During the study, patients were asked about subjective neurologic symptoms such as tingling and numbness. A few patients from all three groups reported such paresthesias from time to time, but they were of a transient nature, not perpetual, and, in patients taking metronidazole, not aggravated despite continuing treatment with unchanged dose.(ABSTRACT TRUNCATED AT 250 WORDS)

Ceftriaxone: pharmacokinetics and effect on the intestinal microflora in patients with acute bacterial infections.

12 patients with acute bacterial infections were treated with ceftriaxone, 1.5 g intravenously twice daily for 7-13 days. Pharmacokinetic variables were studied in 11 patients. In older subjects, serum half-lives were longer and serum clearances lower than in younger individuals. After the last dose, a larger increase in AUC compared to the first dose was observed in older patients and a biphasic elimination curve appeared in all patients but 2, with a terminal half-life of 15.6 h and 11.4 in old and young subjects, respectively. Estimated biliary clearances showed large individual variation, with a range of 0-16 ml/min X 1.73 m2. Changes in the colonic microflora were pronounced. Almost total disappearance of staphylococci, streptococci and enterobacteria was found, and there was a marked tendency to overgrowth of yeasts and enterococci. One patient with the highest estimated biliary clearance of ceftriaxone developed diarrhoea after 7 days of therapy. A toxin-producing Clostridium difficile was isolated from the stool.

Metronidazole: pharmacokinetic observations in severely ill patients.

Single and multiple dose pharmacokinetics of metronidazole and its two major metabolites were studied in ten patients. Patients with hepatic insufficiency had longer average serum half-life of metronidazole (11.2 h) than individuals with normal liver and kidney function (5.9 h) or isolated moderate renal impairment (6.5 h). Patients with hepatic disorder presented larger areas under the serum concentration curve, lower serum clearances and a tendency to more rapidly rising trough values of metronidazole. In patients with renal insufficiency trough values of the hydroxy metabolite seemed to rise faster and serum half-life was prolonged. The acetic acid metabolite was detected in serum of all patients with renal dysfunction but only in half of those with normal renal function and then at lower levels. A reduced 24-h total urinary recovery of metronidazole among patients with renal disorder was explained by a decreased excretion of hydroxy metabolite. The kinetics of metronidazole itself seem not to be influenced by renal impairment while the elimination rate of metabolites is reduced. The decreased clearance of the drug in patients with hepatic dysfunction makes a dose reduction in this patient group advisable.

Infectious complications to granulocytopenia.

A consecutive series of patients with granulocytopenia was analysed with consideration of rate and spectrum of infectious complications. The records of 98 patients from 1975-81 were studied retrospectively. Fever was the most common symptom of infection. 78 patients had one or more infectious manifestations, and Staphylococcus aureus was the most common pathogen isolated. Septicemia occurred in 22 patients of which 17 were caused by gram-negative bacteria. Pseudomonas aeruginosa caused septicemia in 8 patients. The risk of septicemia increased with a decreasing cell count and with the degree and duration of fever. The mortality rate for the whole group was 24% and for the septicemic patients 41%.

A comparative study of metronidazole and sulfasalazine for active Crohn's disease: the cooperative Crohn's disease study in Sweden. I. Design and methodologic considerations.

The design and execution of the Cooperative Crohn's Disease Study in Sweden are described in this paper. A double-blind, double-dummy, crossover (2 X 4 mo) technique was used to compare the suppressive efficacy of metronidazole (0.4 g b.i.d.) and sulfasalazine (1.5 g b.i.d.). The number of randomized patients (78) presented approximately one-third of the available population. The Crohn's Disease Activity Index and the plasma level of orosomucoid were the main variables for clinical evaluation. Results were analyzed primarily in the first treatment period by ranking the clinical outcome of every patient according to a uniform and detailed scheme and applying Wilcoxon nonparametric statistics. The cross-over data only served as additional information. Thirty-six patients had had earlier and mostly positive experience with sulfasalazine. Repeated plasma drug analysis indicated good compliance. The blindness of the trial was tested and appeared satisfactory. The coordination of the trial proceeded as planned. A lack of full conformity in the electroimmunoassay of orosomucoid was taken care of satisfactorily.

A comparative study of metronidazole and sulfasalazine for active Crohn's disease: the cooperative Crohn's disease study in Sweden. II. Result.

Seventy-eight patients with active Crohn's disease participated in a randomized, double-blind, cross-over trial. The study comprised two 4-mo period. The purpose was to test the efficacy of metronidazole in comparison with that of sulfasalazine. As the main evaluation criteria the Crohn's Disease Activity Index and plasma levels of orosomucoid were chosen. In the first period no difference in efficacy as measured by Crohn's Disease Activity Index was found between the treatment groups. The reduction of the plasma orosomucoid level was significantly more pronounced in the metronidazole group. The hemoglobin concentration increased more in this group than in the sulfasalazine group, possibly due to a toxic effect of sulfasalazine. The erythrocyte sedimentation rate decreased similarly with both drugs. In 15 patients who had active disease throughout the first period, Crohn's Disease Activity Index decreased significantly in the second period for those who switched to metronidazole, but not for those who switched to sulfasalazine. After crossover, no apparent further change in Crohn's Disease Activity Index occurred in either of the treatment groups among patients who had responded favorably in the first period. The plasma concentration of orosomucoid increased significantly among the patients in the sulfasalazine group but not in the metronidazole group. It is therefore concluded that metronidazole is slightly more effective than sulfasalazine in the treatment of crohn's disease. It is worthwhile switching the drug regimen from sulfasalazine, when it fails, to metronidazole, but not from metronidazole to sulfasalazine.