Expression of the peptide antibiotic human beta-defensin 1 in cultured gingival epithelial cells and gingival tissue.

Human beta-defensin-1 (hBD-1) is a member of the family of small cationic antimicrobial peptides that have been identified in several mucosal epithelia. Because human gingival epithelium is a site that is constantly challenged by oral microorganisms, we examined the expression of hBD-1 in human gingival epithelial and fibroblast cell cultures and tissue samples. Cell cultures were challenged with cell wall extracts of Porphyromonas gingivalis or Fusobacterium nucleatum, Escherichia coli lipopolysaccharide, tumor necrosis factor alpha, or phorbol myristate acetate.

Can nurse practitioners offer a quality service? An evaluation of a year's work of a nurse led minor injury unit.

Objective: To evaluate activities of a nurse led minor injuries unit based in a community hospital situated eight miles away from its sister district hospital in Crawley to demonstrate whether nurses can provide an alternative service.

Design: "Walk in" service provided by emergency nurse practitioners (ENPs) working within protocols and parameters with supervision from the accident and emergency consultant during the period of May 1995 to April 1996.

Results: Within the 12 month period, ENPs treated 6944 new patients.

Nucleotide sequence of the inter-structural gene region of feline infectious peritonitis virus.

The sequence of the region located between the S and M glycoprotein genes of the 79-1146 strain of feline infectious peritonitis virus (FIPV) is presented. The inter-structural gene region encodes 3 open reading frames (ORFs), termed ORFs 3a, 3b and 4, with nucleotide sequences conforming to the minimum conserved transcription signal upstream of each. An additional ORF, 3x, partially overlaps the 3' end of ORF 3a.

Non-specific currents at fertilisation in sea urchin oocytes.

Using the whole-cell voltage-clamp technique to clamp sea urchin oocytes we show that the fertilising spermatozoon triggers an inward current of -521 +/- 56.7 pA (n = 8) at activation. Simultaneously, the plasma membrane depolarises and the conductance increases from 23.

Regional difference in expression of characteristic abnormality of harlequin ichthyosis in affected fetuses.

Harlequin ichthyosis (HI) is a severe congenital ichthyosis in which newborn infants are covered with a thick plate of stratum corneum. We examined skin specimens from a variety of regions of the body including the scalp, face, tongue, trunk, upper and lower extremities, digits, palms, and soles of three fetuses affected with HI that were diagnosed prenatally. In all the skin regions, characteristic morphological abnormalities (absent or abnormal lamellar granules and intercellular lamellae, lipid inclusions in the cornified cells) were expressed in the late second trimester of the fetal period.

Intracellular pH regulation in the human oocyte.

We have used fluorescence techniques to study the regulation of intracellular pH during fertilization and preimplantation embryo development in human oocytes. The intracellular pH of human oocytes during maturation and fertilization was always 7.4, suggesting that these processes do not involve long-term changes in intracellular pH.

Diagnostic value of antibodies to filaggrin in rheumatoid arthritis.

Objective: To determine the prevalence of antibodies to filaggrin in a cross sectional sample of patients with rheumatoid arthritis (RA).

Methods: Filaggrin from human skin was either extracted with 0.05% Nonidet P-40 and then partially purified by precipitating in ethanol and resuspending in water (Nonidet preparation) or extracted with 9 M urea and then purified by sequential fractionation on a DEAE Sephadex column and on a strong cation exchange column (purified preparation).

Protein phosphatase activity in human keratinocytes cultured from normal epidermis and epidermis from patients with harlequin ichthyosis.

We investigated serine/threonine protein phosphatase (PP) activity and the expression of PP2A during growth and differentiation of epidermal keratinocytes in culture. Keratinocyte PP activity was strongly inhibited by calyculin A and okadaic acid, indicating that the activity was mainly due to PP2A and PP1. The phosphatase activity decreased to about 20% of the initial (day 1) level by the time of confluence and to about 10% at day 7 postconfluence.

Nitric oxide gates fertilization channels in ascidian oocytes through nicotinamide nucleotide metabolism.

In this paper we use the nitric oxide (NO) donor sodium nitroprusside to examine the response of the unfertilised oocyte of the ascidian Ciona intestinalis to nitric oxide. We show that the release of NO triggers an inward current that displays similar properties to the ascidian fertilisation current. Furthermore, the production of NO causes the release of intracellular calcium through a ruthenium-red sensitive mechanism.

Polarized distribution of L-type calcium channels in early sea urchin embryos.

Using the whole cell clamp technique, we have measured calcium-dependent currents and steady-state conductance in early sea urchin blastomeres. The calcium currents in M phase decreased from 8.5 microA/cm2 at the four-cell stage to 5.

Ambient air and its potential effects on conception in vitro.

Incidences of chemical air contamination (CAC) are common in assisted reproductive technology, but not reported in peer review format. Justified fear of car and industrial emissions clearly exists among reproductive specialists, but standards for air contents and gaseous emission limits have not been reported. Here, we describe air sampling methods and assay systems which can be applied to any laboratory or laboratory item.

Distinctive expression of filaggrin and trichohyalin during various pathways of epithelial differentiation.

Filaggrin and trichohyalin are keratin intermediate filament-associated proteins, and are primarily expressed in the granular cells of the epidermis and in the inner root sheath cells of the hair follicles, respectively. These two proteins are, however, occasionally co-expressed in some tissues. To gain more insights into the mechanisms for expression and processing of (pro)filaggrin and trichohyalin during various pathways of epithelial differentiation, we compared their localization by double immunostaining techniques in normal and psoriatic epidermis, tongue filiform papillae and cultured human epidermal keratinocytes.

Harlequin ichthyosis keratinocytes in lifted culture differentiate poorly by morphologic and biochemical criteria.

Harlequin ichthyosis (HI) is a severe congenital ichthyosis in which massively thickened stratum corneum with abnormal barrier function often results in death of affected newborns. Survivors evolve into a severe nonbullous ichthyosiform erythroderma. Previously we have ascertained three biochemical phenotypes of HI, based on abnormal profilaggrin and K6 and K16 expression in epidermis.

A soluble extract from human spermatozoa activates ascidian oocytes.

A soluble extract from human spermatozoa induced calcium oscillations and extrusion of the first polar body when injected into oocytes of the ascidian Ciona intestinalis. The properties of calcium oscillations and time of polar body extrusion precisely mimic oocyte activation induced by C. intestinalis sperm or sperm extracts.

Effect of high-dose chemotherapy on intestinal permeability in humans.

1. Mucositis is a common side-effect of chemotherapy which is difficult to assess except by invasive means such as upper gastrointestinal endoscopy. Differential absorption of mono- and di-saccharides, such as rhamnose and lactulose, is a non-invasive measure of intestinal damage.

Sperm factor: what is it and what does it do?

There are two current hypotheses as to how the spermatozoon triggers the oocyte into activity; a transmembrane receptor mechanism involving G-proteins and a soluble sperm-factor mechanism. In this short review we show that the present data favours the idea of a soluble factor diffusing from the spermatozoon into the oocyte following plasma membrane fusion of the two gametes that triggers calcium release in the oocyte. Two categories of calcium release mechanisms, inositol-1,4,5-triphosphate-induced calcium release (IICR) and calcium induced calcium release (CICR) are found in oocytes from a variety of species and both appear to be activated at fertilization.

Mobilization of predominantly Philadelphia chromosome-negative blood progenitors using cyclophosphamide and rHUG-CSF in early chronic-phase chronic myeloid leukaemia: correlation with Sokal prognostic index and haematological control.

Mobilization of Philadelphia chromosome (Ph) negative blood progenitors was attempted in 23 newly diagnosed chronic myeloid leukaemia (CML) patients using a regimen of cyclophosphamide (CY) 5 g/m2 and rHUG-CSF 150 microg/m2 daily. This regimen was well tolerated with no major adverse events reported. More than 2 x 10(6)/kg CD34+ cells were collected in 21 patients (91%).

Transient expression of epidermal filaggrin in cultured cells causes collapse of intermediate filament networks with alteration of cell shape and nuclear integrity.

Filaggrin is an intermediate filament-associated protein (IFAP) that aggregates epidermal keratin filaments in vitro and is thought to perform a similar function during terminal differentiation in vivo. To test this function in living cells, we transiently expressed constructs encoding human filaggrin in both simple epithelial cells (COS-7) and rat keratinocytes. Scanning laser confocal microscopy showed that filaggrin-positive cells had collapsed keratin and vimentin intermediate filament (IF) networks, and that filaggrin partially co-localized with the IF networks.

Evidence for specific proteolytic cleavage of the N-terminal domain of human profilaggrin during epidermal differentiation.

Profilaggrin is a large phosphoprotein that is expressed in the granular cells of epidermis where it is localized in keratohyalin. It consists of multiple copies of single filaggrin units plus N- and C-terminal sequences that differ from filaggrin. Profilaggrin is dephosphorylated and proteolytically processed during terminal differentiation to yield filaggrin, which associates with keratin intermediate filaments to form macrofibrils in the lower layers of the stratum corneum.

MALDI-MS as a monitor of the purification and folding of synthetic eclosion hormone.

Analogues of the small protein Manduca sexta eclosion hormone (62 amino acids) were synthesized by Fmoc solid-phase methodology. Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) was used to analyze the products of the syntheses and this information was used to design an efficient purification scheme. MALDI-MS was used to monitor the target products through purification and it was also used to monitor folding of the purified materials.