Skin characteristics: normative data for elasticity, erythema, melanin, and thickness at 16 different anatomical locations.

BACKGROUND: The clinical use of non-invasive instrumentation to evaluate skin characteristics for diagnostic purposes and to evaluate treatment outcomes has become more prevalent. The purpose of this study was to generate normative data for skin elasticity, erythema (vascularity), melanin (pigmentation), and thickness across a broad age range at a wide variety of anatomical locations using the Cutometer(®) (6 mm probe), Mexameter(®) , and high-frequency ultrasound in a healthy adult sample. METHODS: We measured skin characteristics of 241 healthy participants who were stratified according to age and gender. Sixteen different anatomical locations were measured using the Cutometer(®) for maximum skin deformation, gross elasticity, and biological elasticity, the Mexameter(®) for erythema and melanin, and high-frequency ultrasound for skin thickness. Standardized measurement procedures were applied for all participants. RESULTS: The means and standard deviations for each measured skin characteristic for females and males across five different age groups (20-29, 30-39, 40-49, 50-59, 60-69, and 70-85 years old) are presented. As previously described, there were variations in skin characteristics across age groups, anatomical locations, and between females and males highlighting the need to use site specific, age and gender matched data when comparing skin characteristics. CONCLUSION: The reported data provides normative data stratified by anatomical location, age, and gender that can be used by clinicians and researchers to objectively determine whether patients' skin characteristics vary significantly from healthy subjects.

A pilot study of a trauma registry at the Georgetown Public Hospital, Georgetown, Guyana

OBJECTIVE: To complete a pilot study to initiate a trauma registry in Guyana. DESIGN AND METHODS: A trauma registry form was developed and customized locally, based on the Kampala trauma score. A convenience sample of trauma patients was collected over a 49 day period. The inclusion criteria were international code of diseases (ICD) 9 codes 800 to 957. The form recorded triage vital signs, Injury severity score (ISS), details of the event, alcohol and drug use. The forms were completed by the treating physician at the time of the encounter and the data was entered in a spreadsheet. RESULTS: Data from 34 patients were analyzed. The most common causes of injuries were due to falls (26.5%) and road traffic accidents (14.8%), with 38% of injuries occurring on roadways. Bony pelvis and extremities (62%) were the most common site of serious injuries, followed by the head/neck/face (15%). Only 3 persons admitted to alcohol use. No IPV (intimate partner violence) cases were detected. There were 4 stab wounds and no guns shot wounds recorded. CONCLUSION: A trauma registry can capture data that can lead to improved trauma care. However, there were numerous limitations, most notably the low number of forms completed compared to the study population. To implement a working trauma registry we will need to capture all cases. Suggestions to increase the number of patients include converting the form into a mobile telephone application or training data entry clerks. Successful implementation would require support from all stakeholders.

Dual frequency comb spectroscopy with a single laser.

We demonstrate a simple scheme for dual frequency comb spectroscopy in which the second frequency comb is generated by propagating the primary pulse train through a dazzler. The two frequency combs are combined behind a Mach-Zehnder interferometer, and the optical spectrum is read out by an rf-spectrum analyzer. The method is applied to record the overtone absorption spectrum of C2H2 (acetylene) in the wavelength region around 1.03 µm. A spectrum with a resolution of 4 cm(-1) is obtained, which compares well with that from the HITRAN database. A simple method for improving the spectral resolution is demonstrated.

Rapid-scan acousto-optical delay line with 34 kHz scan rate and 15 as precision.

An optical fast scan delay exploiting the near-collinear interaction between a train of ultrashort optical pulses and an acoustic wave propagating in a birefringent crystal is introduced. In combination with a femtosecond Er:fiber laser, the scheme is shown to delay few femtosecond pulses by up to 6 ps with a precision of 15 as. A resolution of 5 fs is obtained for a single sweep at a repetition rate of 34 kHz. This value can be improved to 39 as for multiple scans at a total rate of 0.3 kHz.

Self-guided propagation of ultrashort laser pulses in the anomalous dispersion region of transparent solids: a new regime of filamentation.

We report measurements concerning the propagation of femtosecond laser pulses in fused silica with a wavelength at 1.9 µm falling in the negative group velocity dispersion region. Under sub-GW excitation power, stable filaments are observed over several cm showing the emergence of nonspreading pulses both in space and time. At higher excitation powers, one observes first multiple pulse splitting followed by the emergence of the quasispatiotemporal solitary filament. These results are well reproduced by numerical simulations.

Self-referenced spectral interferometry for ultrashort infrared pulse characterization.

We demonstrate for the first time (to our knowledge) characterization of ultrashort IR pulses by self-referenced spectral interferometry. Both sub-55-fs pulses from 1.4 µm to 2 µm and broadband 2.5-cycle pulses at 1.65 µm (13 fs FWHM) are characterized.

Grism compressor for carrier-envelope phase-stable millijoule-energy chirped pulse amplifier lasers featuring bulk material stretcher.

We demonstrate compression of amplified carrier-envelope phase (CEP)-stable laser pulses using paired transmission gratings and high-index prisms, or grisms, with chromatic dispersion matching that of a bulk material pulse stretcher. Grisms enable the use of larger bulk stretching factors and thereby higher energy pulses with lower B-integral in a compact amplifier design suitable for long-term CEP control.

Few-cycle pulse characterization with an acousto-optic pulse shaper.

An acousto-optic pulse shaper has been used to characterize few-cycle pulses generated in a hollow-core fiber. A grism pair precompensates for the dispersion of the acousto-optic crystal, allowing the full pulse-shaping window to be used for replica generation rather than self-compensation. A 9.4 fs pulse was measured, the shortest ever measured with an acousto-optic pulse shaper, to our knowledge.

Single-shot, high-dynamic-range measurement of sub-15 fs pulses by self-referenced spectral interferometry.

We explore theoretically and numerically the temporal contrast limitation of a self-referenced spectral interferometry measurement. An experimental confirmation is given by characterization and fine compression of hollow-core fiber generated sub-15 fs pulses, yielding an accurately measured coherent contrast of 50 dB on a ±400 fs time range.

Bacterial diversity in Fe-rich hydrothermal sediments at two South Tonga Arc submarine volcanoes.

Seafloor iron oxide deposits are a common feature of submarine hydrothermal systems. Morphological study of these deposits has led investigators to suggest a microbiological role in their formation, through the oxidation of reduced Fe in hydrothermal fluids. Fe-oxidizing bacteria, including the recently described Zetaproteobacteria, have been isolated from a few of these deposits but generally little is known about the microbial diversity associated with this habitat. In this study, we characterized bacterial diversity in two Fe oxide samples collected on the seafloor of Volcanoes 1 and 19 on the South Tonga Arc. We were particularly interested in confirming the presence of Zetaproteobacteria at these two sites and in documenting the diversity of groups other than Fe oxidizers. Our results (small subunit rRNA gene sequence data) showed a surprisingly high bacterial diversity, with 150 operational taxonomic units belonging to 19 distinct taxonomic groups. Both samples were dominated by Zetaproteobacteria Fe oxidizers. This group was most abundant at Volcano 1, where sediments were richer in Fe and contained more crystalline forms of Fe oxides. Other groups of bacteria found at these two sites include known S- and a few N-metabolizing bacteria, all ubiquitous in marine environments. The low similarity of our clones with the GenBank database suggests that new species and perhaps new families were recovered. The results of this study suggest that Fe-rich hydrothermal sediments, while dominated by Fe oxidizers, can be exploited by a variety of autotrophic and heterotrophic micro-organisms.

Closed-loop carrier-envelope phase stabilization with an acousto-optic programmable dispersive filter.

We demonstrate arbitrary carrier-envelope (CE) phase control of femtosecond laser pulses by an acousto-optic programmable dispersive filter (AOPDF), with an accuracy better than pi/100 at a repetition rate of 1 kHz. We also demonstrate, for the first time to the best of our knowledge, 15 Hz closed-loop CE phase stabilization using an AOPDF inside a 1 kHz chirped pulse amplifier to correct for slow CE phase drifts.

Scalable Yb-MOPA-driven carrier-envelope phase-stable few-cycle parametric amplifier at 1.5 microm.

Carrier-envelope phase-stable 4 microJ pulses at approximately 1.5 microm are obtained from a femtosecond Yb:KGW-MOPA-pumped two-stage optical parametric amplifier. This novel technology represents a highly attractive alternative to traditional Ti:sapphire front-ends for seeding multimillijoule-level optical parametric chirped-pulse amplifiers. For this task, we demonstrate stretching of the OPA output to approximately 40 ps and recompression to 33 fs pulse duration. As a stand-alone system, our tunable two-stage OPA might find numerous applications in time-resolved spectroscopy and micromachining.

High-dynamic-range temporal measurements ofshort pulses amplified by OPCPA.

We report on the first experimental measurement of high-dynamic-range pulse contrast of compressed optical parametric chirped-pulse-amplification (OPCPA) pulses on the picosecond scale. The measured -80-dB OPCPA contrast at 1054 nm agrees well with theoretical predictions and exceeds the estimated and measured levels for comparable amplification in a Ti:sapphire regenerative amplifier by approximately 10 dB. A key to achieving better contrast with OPCPA is the simpler experimental setup that promotes more-efficient coupling of seed pulse energy into the amplification system.

Parametric amplification of few-cycle carrier-envelope phase-stable pulses at 2.1 microm.

We demonstrate an optical parametric chirped-pulse amplifier producing infrared 20 fs (3-optical-cycle) pulses with a stable carrier-envelope phase. The amplifier is seeded with self-phase-stabilized pulses obtained by optical rectification of the output of an ultrabroadband Ti:sapphire oscillator. Energies of -80 microJ with a well-suppressed background of parametric superfluorescence and up to 400 microJ with a superfluorescence background are obtained from a two-stage parametric amplifier based on periodically poled LiNbO3 and LiTaO3 crystals. The parametric amplifier is pumped by an optically synchronized 1 kHz, 30 ps, 1053 nm Nd:YLF amplifier seeded by the same Ti:sapphire oscillator.

Carboxy-terminal processing of the large subunit of [Fe] hydrogenase from Desulfovibrio desulfuricans ATCC 7757.

hydA and hydB, the genes encoding the large (46-kDa) and small (13. 5-kDa) subunits of the periplasmic [Fe] hydrogenase from Desulfovibrio desulfuricans ATCC 7757, have been cloned and sequenced. The deduced amino acid sequence of the genes product showed complete identity to the sequence of the well-characterized [Fe] hydrogenase from the closely related species Desulfovibrio vulgaris Hildenborough (G. Voordouw and S. Brenner, Eur. J. Biochem. 148:515-520, 1985). The data show that in addition to the well-known signal peptide preceding the NH2 terminus of the mature small subunit, the large subunit undergoes a carboxy-terminal processing involving the cleavage of a peptide of 24 residues, in agreement with the recently reported data on the three-dimensional structure of the enzyme (Y. Nicolet, C. Piras, P. Legrand, E. C. Hatchikian, and J. C. Fontecilla-Camps, Structure 7:13-23, 1999). We suggest that this C-terminal processing is involved in the export of the protein to the periplasm.

[3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis.

The role of the high potential [3Fe-4S]1+,0 cluster of [NiFe] hydrogenase from Desulfovibrio species located halfway between the proximal and distal low potential [4Fe-4S]2+,1+ clusters has been investigated by using site-directed mutagenesis. Proline 238 of Desulfovibrio fructosovorans [NiFe] hydrogenase, which occupies the position of a potential ligand of the lacking fourth Fe-site of the [3Fe-4S] cluster, was replaced by a cysteine residue. The properties of the mutant enzyme were investigated in terms of enzymatic activity, EPR, and redox properties of the iron-sulfur centers and crystallographic structure. We have shown on the basis of both spectroscopic and x-ray crystallographic studies that the [3Fe-4S] cluster of D. fructosovorans hydrogenase was converted into a [4Fe-4S] center in the P238 mutant. The [3Fe-4S] to [4Fe-4S] cluster conversion resulted in a lowering of approximately 300 mV of the midpoint potential of the modified cluster, whereas no significant alteration of the spectroscopic and redox properties of the two native [4Fe-4S] clusters and the NiFe center occurred. The significant decrease of the midpoint potential of the intermediate Fe-S cluster had only a slight effect on the catalytic activity of the P238C mutant as compared with the wild-type enzyme. The implications of the results for the role of the high-potential [3Fe-4S] cluster in the intramolecular electron transfer pathway are discussed.

Heterologous expression of the Desulfovibrio gigas [NiFe] hydrogenase in Desulfovibrio fructosovorans MR400.

The ability of Desulfovibrio fructosovorans MR400 DeltahynABC to express the heterologous cloned [NiFe] hydrogenase of Desulfovibrio gigas was investigated. The [NiFe] hydrogenase operon from D. gigas, hynABCD, was cloned, sequenced, and introduced into D. fructosovorans MR400. A portion of the recombinant heterologous [NiFe] hydrogenase was totally matured, exhibiting catalytic and spectroscopic properties identical to those of the native D. gigas protein. A chimeric operon containing hynAB from D. gigas and hynC from D. fructosovorans placed under the control of the D. fructosovorans hynAp promoter was constructed and expressed in D. fructosovorans MR400. Under these conditions, the same level of activity was obtained as with the D. gigas hydrogenase operon.

Molecular study and partial characterization of iron-only hydrogenase in Desulfovibrio fructosovorans.

An iron-only hydrogenase was partially purified and characterized from Desulfovibrio fructosovorans wild-type strain. The enzyme exhibits a molecular mass of 56 kDa and is composed of two distinct subunits HydA and HydB (46 and 13 kDa, respectively). The N-terminal amino acid sequences of the two subunits of the enzyme were determined with the aim of designing degenerate oligonucleotides. Direct and inverse polymerase chain reaction techniques were used to clone the hydrogenase encoding genes. A 9-nucleotide region located 75 bp upstream from the translational start codon of the D. fructosovorans hydA gene was found to be highly conserved. The analysis of the deduced amino acid sequence of these genes showed the presence of a signal sequence located in the small subunit, exhibiting the consensus sequence which is likely to be involved in the specific export mechanism of hydrogenases. Two ferredoxin-like motives involved in the coordination of [4Fe-4S] clusters were identified in the N-terminal domain of the large subunit. The amino acid sequence of the [Fe] hydrogenase from D. fructosovorans was compared with the amino acid sequences from eight other hydrogenases (cytoplasmic and periplasmic). These enzymes share an overall 18% identity and 28% similarity. The identity reached 73% and 69% when the D. fructosovorans hydrogenase sequence was compared with the hydrogenase sequences from Desulfovibrio vulgaris Hildenborough and Desulfovibrio vulgaris oxamicus Monticello, respectively.

Further characterization of the two tetraheme cytochromes c3 from Desulfovibiro africanus: nucleotide sequences, EPR spectroscopy and biological activity.

The genes encoding the basic and acidic tetraheme cytochromes c3 from Desulfovibrio africanus have been sequenced. The corresponding amino acid sequences of the basic and acidic cytochromes c3 indicate that the mature proteins consist of a single polypeptide chain of 117 and 103 residues, respectively. Their molecular masses, 15102 and 13742 Da, respectively, determined by mass spectrometry, are in perfect agreement with those calculated from their amino acid sequences. Both D. africanus cytochromes c3 are synthesized as precursor proteins with signal peptides of 23 and 24 residues for the basic and acidic cytochromes, respectively. These cytochromes c3 exhibit the main structural features of the cytochrome c3 family and contain the 16 strictly conserved cysteine + histidine residues directly involved in the heme binding sites. The D. africanus acidic cytochrome c3 differs from all the other homologous cytochromes by its low content of basic residues and its distribution of charged residues in the amino acid sequence. The presence of four hemes per molecule was confirmed by EPR spectroscopy in both cytochromes c3. The g-value analysis suggests that in both cytochromes, the angle between imidazole planes of the axial histidine ligands is close to 90 degrees for one heme and much lower for the three others. Moreover, an unusually high exchange interaction (approximately 10[-2] cm[-1]) was evidenced between the highest potential heme (-90 mV) and one of the low potential hemes in the basic cytochrome c3. The reactivity of D. africanus cytochromes c3 with heterologous [NiFe] and [Fe] hydrogenases was investigated. Only the basic one interacts with the two types of hydrogenase to achieve efficient electron transfer, whereas the acidic cytochrome c3 exchanges electrons specifically with the basic cytochrome c3. The difference in the specificity of the two D. africanus cytochromes c3 has been correlated with their highly different content of basic and acidic residues.

Involvement of a single periplasmic hydrogenase for both hydrogen uptake and production in some Desulfovibrio species.

Various sulphate-reducing bacteria differing in the number of genes encoding hydrogenase were shown to ferment lactate in coculture with Methanospirillum hungatei, in the absence of sulphate. The efficiency of interspecies H2 transfer carried out by these species of sulphate-reducing bacteria does not appear to correlate with the distribution of genes coding for hydrogenase. Desulfovibrio vulgaris Groningen, which possesses only the gene for [NiFe] hydrogenase, oxidizes hydrogen in the presence of sulphate and produces some hydrogen during fermentation of pyruvate without electron acceptor. The hydrogenase of D. vulgaris was purified and characterized. It exhibits a molecular mass of 87 kDa and is composed of two different subunits (60 and 28 kDa). D. vulgaris hydrogenase contains 10.6 iron atoms, 0.9 nickel atom and 12 acid-labile sulphur atoms/molecule, and the absorption spectrum of the enzyme is characteristic of an iron-sulphur protein. Maximal H2 uptake and H2 evolution activities were 332 and 230 units/mg protein, respectively. D. vulgaris cells contain exclusively the [NiFe] hydrogenase, whatever the growth conditions, as shown by biochemical and immunological studies. Immunocytolocalization in ultrathin frozen sections of cells grown on lactate and sulphate, on H2 and sulphate and on pyruvate showed that the [NiFe] hydrogenase was located in the periplasmic space. Labelling was enhanced in cells grown on H2 and sulphate and on pyruvate. The results enable us to conclude that D. vulgaris Groningen contains a single hydrogenase of the [NiFe] type, located in the periplasmic space like that described for D. gigas. This enzyme appears to be involved in both H2 uptake and H2 production, depending on the growth conditions.