FAMILIAL VALVULAR PULMONIC STENOSIS INVOLVING THREE SIBLINGS

This report describes the cases of three mentally and physically well-developed siblings, 12, 10, and 8 years of age, with varying degrees of isolated valvular pulmonic stenosis not related to age. The severest lesion occurred in the middle patient and was associated with a right-to-left shunt through a patent foramen ovale. The three children had no other siblings, and there was no history of congenital heart disease among close relatives. These cases support the conclusion that genetic factors play a significant role in the development of certain congenital cardiac disorders.     

Bioaccumulation of Key Metals and Other Contaminants by Seaweeds from the Egyptian Mediterranean Sea Coast in Relation to Human Health Risk

This study examined the bioaccumulation of key metals and other contaminants (SO₄, F, Na, K, Ca, Mg, P, Si, Fe, Mn, Zn, Ni, Co, Cu, and Cd) by seaweeds from the Egyptian Mediterranean Sea coast and their risk to human health. Bioaccumulation factor calculations indicated that S, F, Na, K, Zn, Cu, and Cd had high seaweed uptake and this referred to the potential pollution of the coastal area. Multivariate analysis of the studied contaminants confirmed the bioaccumulation factor results as well as the physiological processes in seaweeds. Human health risk assessment of F, Mn, Zn, Ni, Cu, and Cd was conducted using hazard quotient (HQ) and incremental lifetime cancer risk (ILCR) analyses. The carcinogenic assessment of sediments and ingestion of seaweeds using ILCR calculations revealed that Mn, Ni, and Cd in sediments and F and Zn in seaweeds require remedial action in order to reduce their human health risks. Additionally, HQ calculations for the contaminants in seaweeds and reference to toxicological references values from different agencies revealed that F and Co present high human health risk effects. Consequently, the potential public health risks from dietary exposure to hazardous contaminants in seaweeds must be continually subjected to research, regulation, and debate.     

Exploring the Mechanism of Biocatalyst Inhibition in Microbial Desulfurization

Microbial desulfurization, or biodesulfurization (BDS), of fuels is a promising technology because it can desulfurize compounds that are recalcitrant to the current standard technology in the oil industry. One of the obstacles to the commercialization of BDS is the reduction in biocatalyst activity concomitant with the accumulation of the end product, 2-hydroxybiphenyl (HBP), during the process. BDS experiments were performed by incubating Rhodococcus erythropolis IGTS8 resting-cell suspensions with hexadecane at 0.50 (vol/vol) containing 10 mM dibenzothiophene. The resin Dowex Optipore SD-2 was added to the BDS experiments at resin concentrations of 0, 10, or 50 g resin/liter total volume. The HBP concentration within the cytoplasm was estimated to decrease from 1,100 to 260 μM with increasing resin concentration. Despite this finding, productivity did not increase with the resin concentration. This led us to focus on the susceptibility of the desulfurization enzymes toward HBP. Dose-response experiments were performed to identify major inhibitory interactions in the most common BDS pathway, the 4S pathway. HBP was responsible for three of the four major inhibitory interactions identified. The concentrations of HBP that led to a 50% reduction in the enzymes'' activities (IC₅₀s) for DszA, DszB, and DszC were measured to be 60 ± 5 μM, 110 ± 10 μM, and 50 ± 5 μM, respectively. The fact that the IC₅₀s for HBP are all significantly lower than the cytoplasmic HBP concentration suggests that the inhibition of the desulfurization enzymes by HBP is responsible for the observed reduction in biocatalyst activity concomitant with HBP generation.     

Analysis of intracellular state based on controlled 3D nanostructures mediated surface enhanced Raman scattering

Near-infrared surface-enhanced Raman spectroscopy (SERS) is a powerful technique for analyzing the chemical composition within a single living cell at unprecedented resolution. However, current SERS methods employing uncontrollable colloidal metal particles or non-uniformly distributed metal particles on a substrate as SERS-active sites show relatively low reliability and reproducibility. Here, we report a highly-ordered SERS-active surface that is provided by a gold nano-dots array based on thermal evaporation of gold onto an ITO surface through a nanoporous alumina mask. This new combined technique showed a broader distribution of hot spots and a higher signal-to-noise ratio than current SERS techniques due to the highly reproducible and uniform geometrical structures over a large area. This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes. We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells. The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells. This SERS technique employing a tightly controlled nanostructure array can potentially be applied to single cell analysis, early cancer diagnosis and cell physiology research.     

Functional genomics by NMR spectroscopy. Phenylacetate catabolism in Escherichia coli.

Aerobic metabolism of phenylalanine in most bacteria proceeds via oxidation to phenylacetate. Surprisingly, the further metabolism of phenylacetate has not been elucidated, even in well studied bacteria such as Escherichia coli. The only committed step is the conversion of phenylacetate into phenylacetyl-CoA. The paa operon of E. coli encodes 14 polypeptides involved in the catabolism of phenylacetate. We have found that E. coli K12 mutants with a deletion of the paaF, paaG, paaH, paaJ or paaZ gene are unable to grow with phenylacetate as carbon source. Incubation of a paaG mutant with [U-13C8]phenylacetate yielded ring-1,2-dihydroxy-1,2-dihydrophenylacetyl lactone as shown by NMR spectroscopy. Incubation of the paaF and paaH mutants with phenylacetate yielded delta3-dehydroadipate and 3-hydroxyadipate, respectively. The origin of the carbon atoms of these C6 compounds from the aromatic ring was shown using [ring-13C6]phenylacetate. The paaG and paaZ mutants also converted phenylacetate into ortho-hydroxyphenylacetate, which was previously identified as a dead end product of phenylacetate catabolism. These data, in conjunction with protein sequence data, suggest a novel catabolic pathway via CoA thioesters. According to this, phenylacetyl-CoA is attacked by a ring-oxygenase/reductase (PaaABCDE proteins), generating a hydroxylated and reduced derivative of phenylacetyl-CoA, which is not re-oxidized to a dihydroxylated aromatic intermediate, as in other known aromatic pathways. Rather, it is proposed that this nonaromatic intermediate CoA ester is further metabolized in a complex reaction sequence comprising enoyl-CoA isomerization/hydration, nonoxygenolytic ring opening, and dehydrogenation catalyzed by the PaaG and PaaZ proteins. The subsequent beta-oxidation-type degradation of the resulting CoA dicarboxylate via beta-ketoadipyl-CoA to succinyl-CoA and acetyl-CoA appears to be catalyzed by the PaaJ, PaaF and PaaH proteins.     

Biochemical and molecular characterization of phenylacetate-coenzyme A ligase, an enzyme catalyzing the first step in aerobic metabolism of phenylacetic acid in Azoarcus evansii

Phenylacetate-coenzyme A ligase (PA-CoA ligase; AMP forming, EC 6.2. 1.30), the enzyme catalyzing the first step in the aerobic degradation of phenylacetate (PA) in Azoarcus evansii, has been purified and characterized. The gene (paaK) coding for this enzyme was cloned and sequenced. The enzyme catalyzes the reaction of PA with CoA and MgATP to yield phenylacetyl-CoA (PACoA) plus AMP plus PPi. The enzyme was specifically induced after aerobic growth in a chemically defined medium containing PA or phenylalanine (Phe) as the sole carbon source. Growth with 4-hydroxyphenylacetate, benzoate, adipate, or acetate did not induce the synthesis of this enzyme. This enzymatic activity was detected very early in the exponential phase of growth, and a maximal specific activity of 76 nmol min(-1) mg of cell protein(-1) was measured. After 117-fold purification to homogeneity, a specific activity of 48 micromol min(-1) mg of protein(-1) was achieved with a turnover number (catalytic constant) of 40 s(-1). The protein is a monomer of 52 kDa and shows high specificity towards PA; other aromatic or aliphatic acids were not used as substrates. The apparent K(m) values for PA, ATP, and CoA were 14, 60, and 45 microM, respectively. The PA-CoA ligase has an optimum pH of 8 to 8.5 and a pI of 6.3. The enzyme is labile and requires the presence of glycerol for stabilization. The N-terminal amino acid sequence of the purified protein showed no homology with other reported PA-CoA ligases. The gene encoding this enzyme is 1, 320 bp long and codes for a protein of 48.75 kDa (440 amino acids) which shows high similarity with other reported PA-CoA ligases. An amino acid consensus for an AMP binding motif (VX2SSGTTGXP) was identified. The biochemical and molecular characteristics of this enzyme are quite different from those of the isoenzyme catalyzing the same reaction under anaerobic conditions in the same bacterium.     

Fluoride,Some Selected Elements,Lipids, and Protein in the Muscle and Liver Tissues of Five Fish Species along the Egyptian Mediterranean Sea Coast

Fish is a vital, healthy source of animal protein and vitamins. This study was oriented to estimate fluoride, some selected elements, lipids, and protein concentrations in fish. Five fish species were collected from the Egyptian Mediterranean Sea coast during April 2007. Variable content of fluoride, calcium, magnesium, sulphate, phosphorus, lipids, and protein were determined in muscle and liver of fish samples and yielded average values in fish muscle of 11.0 ± 2.0 μg/g, 6.5 ± 3.8 mg/g, 100.2 ± 39.3 mg/g, 4.4 ± 6.3 mg/g, 2.4 ± 0.5 mg/g, 6.0 ± 2.2 mg/l, and 2.6 ± 1.0 g% wet wt, respectively. Special attention was given to fluoride because of its hazardous classification. Interestingly, the hazard index values of fluoride were less than 1 for all collected fish species. Additionally, the daily fluoride exposure was generally below the nutrient reference values for Australian and New Zealand populations. Accordingly, there is little human health risk from the selected fish species due to consumption of fluoride-contaminated fish.     

ECHOCARDIOGRAPHIC FINDINGS IN BACTERIAL ENDOCARDITIS

Fifteen echocardiographic recordings in nine patients with bacterial endocarditis revealed vegetations in six cases. The vegetations appeared as uneven, irregular thickening of a valve, a mass of shaggy, dense echoes attached to a leaflet or cusp, or a mass of irregular dense echoes in the cavity or outflow tract of the left ventricle. Such findings were seen only on the echocardiograms of very sick patients with severe valvular lesions. Three patients had flail mitral valves. Echocardiography was not helpful in differentiating between active and healed lesions. Problems in the identification and differential diagnosis of vegetations shown on echocardiograms are discussed.     

Neurotherapeutic Effects of Bee Venom in a Rotenone-Induced Mouse Model of Parkinson’s Disease

Neurophysiology - Parkinson’s disease (PD) is a widespread progressive neurodegenerative disease; its main neuropathological hallmark is massive loss of dopaminergic neurons. Most PD studies...