Role of embB in natural and acquired resistance to ethambutol in mycobacteria

The mycobacterial embCAB operon encodes arabinosyl transferases, putative targets of the antimycobacterial agent ethambutol (EMB). Mutations in embB lead to resistance to EMB in Mycobacterium tuberculosis. The basis for natural, intrinsic resistance to EMB in nontuberculous mycobacteria (NTM) is not known; neither is the practical implication of resistance to EMB in the absence of embB mutations in M. tuberculosis well understood. The conserved embB resistance-determining region (ERDR) of a collection of 13 strains of NTM and 12 EMB-resistant strains of M. tuberculosis was investigated. Genotypes were correlated with drug susceptibility phenotypes. High-level natural resistance to EMB (MIC, . or =64 microg/ml) was associated with a variant amino acid motif in the ERDR of M. abscessus, M. chelonae, and M. leprae. Transfer of the M. abscessus emb allele to M. smegmatis resulted in a 500-fold increase in the MICs. In M. tuberculosis, embB mutations were associated with MICs of > or =20 microg/ml while resistance not associated with an ERDR mutation generally resulted in MICs of < or =10 microg/ml. These data further support the notion that the emb region determines intrinsic and acquired resistance to EMB and might help in the reassessment of the current recommendations for the screening and treatment of infections with EMB-resistant M. tuberculosis and NTM.     

Visualization of mitochondria with green fluorescent protein in cultured fibroblasts from patients with mitochondrial diseases

cDNAs for green fluorescent protein (GFP) and for a GFP fusion protein containing the presequence of human ornithine transcarbamylase (pOTC-GFP) were transfected into cultured human fibroblasts. GFP cDNA gave diffuse fluorescence throughout the cytoplasm and the nucleus, whereas pOTC-GFP cDNA gave mitochondria-associated fluorescence. Fluorescent mitochondrial structures could be classified into five patterns: thread-like mitochondria, fine thread-like ones, rod-like ones, granular ones, and granular ones with weak cytosolic fluorescence. pOTC-GFP mutants resulted in a loss of mitochondrial fluorescence and an appearance of weak fluorescence throughout the cytoplasm. pOTC-GFP cDNA was transfected into fibroblasts from patients with various mitochondrial diseases. Higher ratios of fibroblasts with granular mitochondria and those with fine thread-like ones were observed in a patient with Reye's syndrome and a patient with Kearns-Sayre syndrome. Weak cytosolic fluorescence was sometimes observed in fibroblasts from these patients. This method will be useful to analyze mitochondrial structural alterations and disorders of mitochondrial protein import.     

Long-term efficacy of cyclical etidronate therapy in postmenopausal osteoporosis

Sixty-two women (mean age 68.7 +/- 0.9 yr) with postmenopausal spinal osteoporosis were treated with cyclical etidronate therapy (400 mg for 2 weeks alternating with 12 weeks of 1 gm elemental calcium and 400 IU Vitamin D3) for a minimum of 2 yr. Bone mineral density (BMD) of the lumbar spine (g/cm2) increased significantly (p < 0.0001) after yr 1 (4.1 +/- 0.5 per cent) and yr 2 compared with yr 1 (2.2 +/- 0.5 per cent). The response rate was 89 per cent after yr 1 and 84 per cent after yr 2. BMD of the hip (30 patients) increased by 1.5 +/- 0.9 per cent after yr 1 and 5.5 +/- 1.1 per cent (p < 0.0001) after yr 2 when compared with baseline. The response rate was 63 per cent after yr 1 and 80 per cent after yr 2. Smaller numbers of patients continued with treatment up to 4 yr with no adverse long-term effects.     

Human recombinant anti-Rh(D) monoclonal antibodies: improvement of biological properties by in vitro class-switch

The aim of this study was to investigate how intrauterine growth retardation affects body proportions in VLBW infants. The cohort consisted of 135 surviving and 80 deceased preterm infants weighing less than 1250 grams at birth. Gestational age varied between 24 and 36 weeks (mean age 29.7 and 27.5 weeks, respectively). Birth weight was more than 2 SD below the mean birth standard values in 32% of the surviving, and in 27% of the deceased infants. Reduction of weight, length and head circumference at birth was analysed using Z scores based on Swedish birth standards. Z scores of weight, length and head circumference were highly correlated in the surviving and the deceased infants (r = 0.78 to 0.94 and 0.65 to 0.97, respectively). Length was significantly more affected by growth retardation than weight. Weight and head circumference were proportionately reduced. Intrauterine growth retardation influences body proportions in VLBW infants differently than in larger preterm and term infants.     

Electrical linewidth test structures fabricated in monocrystallinefilms for reference-material applications

The physical widths of reference features incorporated into electrical linewidth test structures patterned in films of monocrystalline silicon have been determined from Kelvin voltage measurements. The films in which the test structures are patterned are electrically insulated from the bulk-silicon substrate by a layer of silicon dioxide provided by SIMOX (Separation by the IMplantation of OXygen) processing. The motivation is to facilitate the development of linewidth reference materials for critical-dimension (CD) metrology-instrument calibration. The selection of the (110) orientation of the starting silicon and the orientation of the structures' features relative to the crystal lattice enable a lattice-plane-selective etch to generate reference-feature properties of rectangular cross section and atomically planar sidewalls. These properties are highly desirable for CD applications in which feature widths are certified with nanometer-level uncertainty for use by a diverse range of CD instruments. End applications include the development and calibration of new generations of CD instruments directed at controlling processes for manufacturing devices having sub-quarter-micrometer features     

A unified diode model for circuit simulation

A new commercially available diode model is described. This unified model is capable of simulating the widest range of diode technologies of any presently available. The emphasis of this paper is on describing the model's extensive features and flexibility in the different domains of operation and is of particular interest in power applications     

National survey of blindness and low vision in Lebanon

Here we report a case that presented with sudden onset of neurological symptoms and was treated with ganciclovir. Polymerase chain reaction analysis for human herpes virus-6 (HHV-6) from his cerebrospinal fluid was later found to be positive. He subsequently recovered with minimal residual neurological symptoms. Encephalitis secondary to this virus may be more common than previously thought in patients undergoing bone marrow transplantation. This condition should be considered in the differential diagnosis of sudden onset neurological symptoms after bone marrow transplantation.     

Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number

The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined. DNA sequence analysis revealed four putative coding regions (open reading frame A [ORFA], ORFA2, ORFB, and ORFC). ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C. glutamicum) and pNG2 (from Corynebacterium diphtheriae). This function was confirmed by deletion mapping of the minimal replicon of pGA1 (1.7 kb) which contains only ORFC. Deletion derivatives of pGA1 devoid of ORFA exhibited significant decreases in the copy number in C. glutamicum cells and displayed segregational instability. Introduction of ORFA in trans into the cells harboring these deletion plasmids dramatically increased their copy number and segregational stability. The ORFA gene product thus positively influences plasmid copy number. This is the first report on such activity associated with a nonintegrating bacterial plasmid. The related plasmids pGA1, pSR1, and pNG2 lacking significant homology with any other plasmid seem to be representatives of a new group of plasmids replicating in the rolling-circle mode.