Regulation of colonic apical potassium (BK) channels by cAMP and somatostatin.

High-conductance apical K+ (BK) channels are present in surface colonocytes of mammalian (including human) colon. Their location makes them well fitted to contribute to the excessive intestinal K(+) losses often associated with infective diarrhea. Since many channel proteins are regulated by phosphorylation, we evaluated the roles of protein kinase A (PKA) and phosphatases in the modulation of apical BK channel activity in surface colonocytes from rat distal colon using patch-clamp techniques, having first increased channel abundance by chronic dietary K+ enrichment. We found that PKA activation using 50 micromol/l forskolin and 5 mmol/l 3-isobutyl-1-methylxanthine stimulated BK channels in cell-attached patches and the catalytic subunit of PKA (200 U/ml) had a similar effect in excised inside-out patches. The antidiarrheal peptide somatostatin (SOM; 2 micromol/l) had a G protein-dependent inhibitory effect on BK channels in cell-attached patches, which was unaffected by pretreatment with 10 micromol/l okadaic acid (an inhibitor of protein phosphatase type 1 and type 2A) but completely prevented by pretreatment with 100 micromol/l Na+ orthovanadate and 10 micromol/l BpV (inhibitors of phosphoprotein tyrosine phosphatase). SOM also inhibited apical BK channels in surface colonocytes in human distal colon. We conclude that cAMP-dependent PKA activates apical BK channels and may enhance colonic K+ losses in some cases of secretory diarrhea. SOM inhibits apical BK channels through a phosphoprotein tyrosine phosphatase-dependent mechanism, which could form the basis of new antidiarrheal strategies.

Altered cryptal expression of luminal potassium (BK) channels in ulcerative colitis.

Decreased sodium (Na(+)), chloride (Cl(-)), and water absorption, and increased potassium (K(+)) secretion, contribute to the pathogenesis of diarrhoea in ulcerative colitis. The cellular abnormalities underlying decreased Na(+) and Cl(-) absorption are becoming clearer, but the mechanism of increased K(+) secretion is unknown. Human colon is normally a K(+) secretory epithelium, making it likely that K(+) channels are expressed in the luminal (apical) membrane. Based on the assumption that these K(+) channels resembled the high conductance luminal K(+) (BK) channels previously identified in rat colon, we used molecular and patch clamp recording techniques to evaluate BK channel expression in normal and inflamed human colon, and the distribution and characteristics of these channels in normal colon. In normal colon, BK channel alpha-subunit protein was immunolocalized to surface cells and upper crypt cells. By contrast, in ulcerative colitis, although BK channel alpha-subunit protein expression was unchanged in surface cells, it extended along the entire crypt irrespective of whether the disease was active or quiescent. BK channel alpha-subunit protein and mRNA expression (evaluated by western blotting and real-time PCR, respectively) were similar in the normal ascending and sigmoid colon. Of the four possible beta-subunits (beta(1-4)), the beta(1)- and beta(3)-subunits were dominant. Voltage-dependent, barium-inhibitable, luminal K(+) channels with a unitary conductance of 214 pS were identified at low abundance in the luminal membrane of surface cells around the openings of sigmoid colonic crypts. We conclude that increased faecal K(+) losses in ulcerative colitis, and possibly other diseases associated with altered colonic K(+) transport, may reflect wider expression of luminal BK channels along the crypt axis.

It ain't over till it's ova: germline sex determination in C. elegans.

Sex determination in most organisms involves a simple binary fate choice between male or female development; the outcome of this decision has profound effects on organismal biology, biochemistry and behaviour. In the nematode C. elegans, there is also a binary choice, either male or hermaphrodite. In C. elegans, distinct genetic pathways control somatic and germline sexual cell fate. Both pathways share a common set of globally acting regulatory genes; however, germline-specific regulatory genes also participate in the decision to make male or female gametes. The determination of sexual fate in the germline of the facultative hermaphrodite poses a special problem, because first sperm then oocytes are produced. It has emerged that additional layers of post-transcriptional regulation have been imposed to modulate the activities of the global sex-determining genes, tra-2 and fem-3; the balance between these activities is crucial in controlling sexual cell fate in the hermaphrodite germline.

Foot compartment syndrome.

FCS is a recognized clinical entity that has few consistent clinical signs except tense swelling. A high degree of clinical suspicion is necessary to provide appropriate treatment. Invasive direct pressure monitoring is needed to diagnose FCS. High-energy injuries are known to cause FCS, but individual risk factors, such as prolonged venous occlusion and blood dyscrasias, are causative factors.

Asymmetrically distributed oligonucleotide repeats in the Caenorhabditis elegans genome sequence that map to regions important for meiotic chromosome segregation.

The roundworm Caenorhabditis elegans has a haploid karyotype containing six linear chromosomes. The termini of worm chromosomes have been proposed to play an important role in meiotic prophase, either when homologs are participating in a genome-wide search for their proper partners or in the initiation of synapsis. For each chromosome one end appears to stimulate crossing-over with the correct homolog; the other end lacks this property. We have used a bioinformatics approach to identify six repetitive sequence elements in the sequenced C.elegans genome whose distribution closely parallels these putative meiotic pairing centers (MPC) or homolog recognition regions (HRR). We propose that these six DNA sequence elements, which are largely chromosome specific, may correspond to the genetically defined HRR/MPC elements.

Fast ignition by intense laser-accelerated proton beams.

The concept of fast ignition with inertial confinement fusion (ICF) is a way to reduce the energy required for ignition and burn and to maximize the gain produced by a single implosion. Based on recent experimental findings at the PETAWATT laser at Lawrence Livermore National Laboratory, an intense proton beam to achieve fast ignition is proposed. It is produced by direct laser acceleration and focused onto the pellet from the rear side of an irradiated target and can be integrated into a hohlraum for indirect drive ICF.

Fabrication of large-aperture lightweight diffractive lenses for use in space.

We describe the advantages of using diffractive (Fresnel) lenses on thin membranes over conventional optics for, among others, future space telescope projects. Fabrication methods are presented for lenses on two types of freestanding membrane up to 50 cm in size. The first is a Fresnel lens etched into a thin (380-microm) glass sheet, and the second is an approximately 50-microm-thick polymer membrane containing a Fresnel lens made by replication process from a specially made fused-silica master. We show optical performance analysis of all the lenses that are fabricated, including a diffraction-limited Airy spot from a 20-m- focal-length membrane lens in a diffractive telescope system.

Heterotopic ossification of the adductor longus muscle presenting as dyspareunia.

Dyspareunia after heterotopic ossification of the adductor longus is a rare complication. We describe a patient with symptomatic heterotopic ossification of the adductor muscle that developed years after sustaining a fracture of the inferior pubic ramus in association with an injury to the adductor longus muscle. The patient's pain was reduced and his dyspareunia resolved after excision of the adductor longus heterotopic ossification and subsequent physical therapy.

Intense high-energy proton beams from Petawatt-laser irradiation of solids.

An intense collimated beam of high-energy protons is emitted normal to the rear surface of thin solid targets irradiated at 1 PW power and peak intensity 3x10(20) W cm(-2). Up to 48 J ( 12%) of the laser energy is transferred to 2x10(13) protons of energy >10 MeV. The energy spectrum exhibits a sharp high-energy cutoff as high as 58 MeV on the axis of the beam which decreases in energy with increasing off axis angle. Proton induced nuclear processes have been observed and used to characterize the beam.

Search for a Caenorhabditis elegans FMR1 homologue: identification of a new putative RNA-binding protein (PRP-1) that hybridizes to the mouse FMR1 double K homology domain.

A mixed stage lambdagt10 Caenorhabditis elegans cDNA library was screened with a probe derived by polymerase chain reaction from the double K homology (KH) domain of mouse FMR1 cDNA, a region that is highly conserved in the human, mouse, chicken, and frog FMR1 proteins. Four positively hybridizing cDNAs were cloned and characterized by sequencing. The overlapping sequences map to cosmid R119 from C. elegans linkage group (chromosome) I, and encode a novel proline-, polyglutamine-, and RGG box-rich putative RNA-binding protein. While the cDNA has two regions with similarity to the mouse double KH domain probe at the nucleotide level, there is no significant similarity of the amino acid sequence with human FMR1, FXR1 or FXR2, nor with KH amino acid motifs. The R119 protein, therefore, does not represent an FMR1 homologue.

High-intensity third-harmonic generation in beta barium borate through second-order and third-order susceptibilities.

The azimuthal dependence of third-order and cascaded second-order nonlinear coupling was used to measure the relative contributions of each to direct third-harmonic generation in beta-barium borate. This permitted the measurement of the values of tensor elements chi(3)(10) , chi(3)(11) , and chi(3)(16) relative to the known chi(2)(ij) . Finally, conversion efficiencies to 3 omega of up to 6% were achieved with a femtosecond chirped-pulse amplification laser with 200 GW/cm(2) in collimated beams.

Petawatt laser pulses.

We have developed a hybrid Ti:sapphire-Nd:glass laser system that produces more than 1500 TW (1.5 PW) of peak power. The system produces 660 J of power in a compressed 440+/-20 fs pulse by use of 94-cm master diffraction gratings. Focusing to an irradiance of >7x10(20) W/cm (2) is achieved by use of a Cassegrainian focusing system employing a plasma mirror.

Iron metabolism and human ferritin heavy chain cDNA from adult brain with an elongated untranslated region: new findings and insights.

Ferritin is a ubiquitous protein which plays a major role in iron sequestration, detoxification and storage. In this paper we highlight the role of ferritin in iron homeostasis and describe factors and diseases that affect its expression. We also describe new studies which further characterize the structure and expression of a novel form of ferritin heavy (H) chain mRNA that was identified in brain and discuss possible implications of these findings. Human fetal and adult brain cDNA libraries previously were screened with cDNA for well-characterized liver ferritin H. In addition to 'liver-like' brain ferritin H cDNA, novel ferritin H cDNAs with an additional 279 nucleotide sequence at the 3'untranslated region (UTR) were identified in both libraries (see refs. 1 and 2; Dhar, M., Chauthaiwale, V., and Joshi, J. G., Gene, 1993, 126, 275 and Dhar, M., and Joshi, J. G., J. Neurochem., 1993, 61, 2140). However, relative to liver ferritin H cDNA, these novel cDNAs were incomplete at their 5'ends [see ref. 3; Joshi, J. G., Fleming, J. T., Dhar, M. S., and Chauthaiwale, V., J. Neurol Sci., 1995, 134, (Suppl.), 52]. In the present paper, by sequencing of cDNAs using reverse transcriptase polymerase chain reaction, we show that the 279 nt 3'UTR sequence, a coding sequence identical to that in human liver ferritin H, and a full-length 5'UTR that includes one mRNA regulatory iron-response element sequence, co-exist in at least one species of ferritin H transcript in six normal human adult and six late-onset, sporadic Alzheimer disease (AD) brains. This sequence is the same in the normal and AD brains. Dot-blot analysis of poly A+ RNAs from different human tissues indicates that relative to the coding sequence of ferritin H, expression of the 279 nt 3'UTR sequence varies among different tissues, is highest in the adult brain, and is very low in fetal brain. In normal adult hippocampus, ferritin H RNA with the novel 279 nt sequence localizes strongly to small non-neuronal cells, capillary endothelial cells, and to selected populations of neurons (granule cells of the dentate gyrus). Significant homology was observed between a region in the 279 nt 3'UTR segment of ferritin H RNA and the 3'UTR of cyclooxygenase-2 mRNA (an inducible iron-containing enzyme involved in prostaglandin synthesis). Possible functions for ferritin H protein derived from the novel message and for the elongated 3'UTR and 5'UTR are discussed.

High-efficiency fused-silica transmission gratings.

We describe the design, fabrication, and performance of high-efficiency transmission gratings fabricated in bulk fused silica for use in high-power ultraviolet laser systems. The gratings exhibit a diffraction efficiency of 94% in order m=-1 and a damage threshold greater than 13>J/cm( 2) for 3-ns pulses at 351 nm. Model calculations and experimental measurements are in good agreement.

125-TW Ti:sapphire/Nd:glass laser system.

We have demonstrated a Ti:sapphire/Nd:glass laser system that produces up to 51 J of energy in 395-fs pulses (125TW). Focusing at f/3 to a 2.5-times diffraction-limited spot results in a peak irradiance greater than 10(20) W/cm(2) . Our 40-cm-diameter gold diffraction gratings have a damage threshold of 0.42 J/cm(2) for 320-fs pulses.

Ablation of human nail by pulsed lasers.

BACKGROUND AND OBJECTIVE: The hard and resistant structure of the nail plate forms a natural barrier that limits the penetration of topical drugs. To overcome this barrier, the use of pulsed laser systems has been suggested. The purpose of this study was to evaluate the effect of four laser systems on nail plate ablation rates, ablation efficiencies, and subsequent craters morphology. STUDY DESIGN/MATERIAL AND METHODS: Solid state Er:YAG (2.94 microns, 250 microseconds), a Ho:YSGG (2.08 microns, 250 microseconds), a XeC1 Excimer (308 nm, 15 ns), and a novel solid-state ultrashort pulse laser (1.05 microns, 350 fs) were used. Ablation rates, surface morphology, and extent of collateral damage were evaluated using light and electron microscopy. RESULTS: Best ablation efficiencies were demonstrated with the ultrashort pulsed laser (1 micron/mJ), whereas maximum material removal per pulse was obtained with the Er:YAG laser (80 microns/ pulse). Scanning electron microscopy showed cracking damage with both Ho:YSGG and Er:YAG. XeC1 and the ultrashort pulse system left tissue surfaces free of cracks or thermal damage. CONCLUSION: With its minimal acoustical and mechanical impact, high efficiency, and negligible collateral damage, the ultrashort pulse laser at 3 J/cm2 was found to be the optimal laser system for nail ablation.

Universal grating design for pulse stretching and compression in the 800-1100-nm range.

We have developed a holographically produced master metallic grating that achieves >91% diffraction efficiency over the wavelength range 800-1100 nm and a maximum diffraction efficiency at 1053 nm greater than 93% when used with TM polarization near the Littrow angle. The near-uniform diffraction efficiency with laser wavelength makes this design attractive for use in chirped-pulse amplification systems employing Ti:sapphire, Cr:LiSAF, or Nd:glass and permits high-fidelity stretching and compression of extremely short (10-fs) pulses.

Designing fully continuous phase screens for tailoring focal-plane irradiance profiles.

An iterative algorithm for constructing fully continuous phase screens for tailoring far-field intensity profiles is presented. The algorithm is robust, stable, and, if run properly, maintains the continuous nature of the phase throughout the iterative process. The iterative procedure is applied to generate continuous phase screens to produce a 12th-power super-Gaussian far-field intensity profile.

High-efficiency multilayer dielectric diffraction gratings.

The design and performance of a new type of high-efficiency diffraction grating for use in either transmission or reflection are described. The gratings are produced in a multilayer dielectric coating deposited upon optically flat substrates. By proper design of the multilayer stack and grating structure, a diffraction efficiency in excess of 96% for polarized light in the m = -1 order in reflection has been achieved.

Amplification of femtosecond pulses to 1 J in Cr:LiSrAlF(6).

We have developed a large-aperture, flash-lamp-pumped Cr:LiSrAlF(6) (Cr:LiSAF) amplifier for use in a femtosecond chirped-pulse amplifier system. Optimum design of the 19-mm-diameter amplifier results in a single-pass gain of 5 with good beam quality. This amplifier produces 1.05-J pulses after compression, with a width of 125 fs at a repetition rate of 0.05 Hz.