Bilateral Necrotizing Fasciitis of the Foot Associated with Group B Streptococcus.

Necrotizing fasciitis (NF) is a severe bacterial infection involving fascia and subcutaneous tissue. It generally affects upper or lower extremities unilaterally, and there are few reports of bilateral-extremity NF. Here, we report a case of a 43-year-old male with type 1 diabetes who had NF on the left foot and subsequently developed NF on the other foot 1 week later. The patient survived with antimicrobial therapy and bilateral below-knee amputation. As group B streptococcus (GBS) was isolated by blood culture and culture of excised tissues of both feet, bilateral GBS NF of the foot was diagnosed. GBS is a rare causative pathogen in NF; however, there have been two case reports of bilateral GBS NF of an extremity in which NF appeared on the opposite extremity 1 week after the primary site infection, as in our case. GBS was isolated from cultures of blood and excised tissues of both extremities in both cases. Together, these observations suggest that GBS has a potential to cause secondary NF at remote sites by hematogenous dissemination with approximately 1 week delay and thereby lead to bilateral NF.

Repeated-batch production of galactooligosaccharides from lactose at high concentration by using alginate-immobilized cells of Sporobolomyces singularis YIT 10047.

We investigated the production of galactooligosaccharides (GOS) from lactose by alginate-immobilized cells of Sporobolomyces singularis YIT 10047 (IM-SS). beta-Galactosidase activity was stable at 30 to 50 degrees C but decreased dramatically between 50 and 60 degrees C and disappeared at 70 degrees C in acetate buffer. The enzyme activity remaining was no more than 20% of that of unheated samples after incubation in distilled water at 55 degrees C, whereas its value was about 60% at the same temperature under buffered condition. However, activity was maintained more than 80% with 10% to 50% (w/w) lactose after incubation at 55 degrees C without buffer. In a single-batch reaction, GOS yield was 41.0% with free cells and 40.4% with IM-SS. We attempted a repeated-batch reaction using IM-SS with 600 g L(-1)lactose. IM-SS produced GOS stably for 20 batches (22 h/batch, 440 h in total) at 55 degrees C and pH 5.0 or 6.0. IM-SS produced GOS at 242 g L(-1), at a rate of 8.72 g L(-1) h(-1). Both GOS yield and production rate were higher than those in published experiments on GOS production using immobilized biocatalysts. The repeated-batch reaction with IM-SS would be an ideal system for GOS production because of its stability and high productivity.

Single nucleotide polymorphism-based genome-wide linkage analysis in Japanese atopic dermatitis families.

BACKGROUND: Atopic dermatitis develops as a result of complex interactions between several genetic and environmental factors. To date, 4 genome-wide linkage studies of atopic dermatitis have been performed in Caucasian populations, however, similar studies have not been done in Asian populations. The aim of this study was to identify chromosome regions linked to atopic dermatitis in a Japanese population. METHODS: We used a high-density, single nucleotide polymorphism genotyping assay, the Illumina BeadArray Linkage Mapping Panel (version 4) comprising 5,861 single nucleotide polymorphisms, to perform a genome-wide linkage analysis of 77 Japanese families with 111 affected sib-pairs with atopic dermatitis. RESULTS: We found suggestive evidence for linkage with 15q21 (LOD = 2.01, NPL = 2.87, P = .0012) and weak linkage to 1q24 (LOD = 1.26, NPL = 2.44, P = .008). CONCLUSION: We report the first genome-wide linkage study of atopic dermatitis in an Asian population, and novel loci on chromosomes 15q21 and 1q24 linked to atopic dermatitis. Identification of novel causative genes for atopic dermatitis will advance our understanding of the pathogenesis of atopic dermatitis.

Selective impairment of Toll-like receptor 2-mediated proinflammatory cytokine production by monocytes from patients with atopic dermatitis.

BACKGROUND: The skin of patients with atopic dermatitis (AD) exhibits a striking susceptibility to infection with gram-positive bacteria and herpes simplex virus (HSV), which are known to stimulate Toll-like receptor (TLR) 2. OBJECTIVE: We investigated whether TLR2-mediated proinflammatory cytokine production by monocytes is selectively impaired in patients with AD and, if so, whether high FcvarepsilonRI levels on the monocytes could be related to the impairment. METHODS: The 2 subpopulations of monocytes, CD14(dim) proinflammatory and CD14(bright) classical monocytes, from patients with AD and healthy control subjects were stimulated to produce IL-1beta and TNF-alpha with phorbol 12-myristate 13-acetate/ionomycin, LPS (TLR4 ligand), or Pam3Cys (TLR2 ligand) for 4 hours, and simultaneous flow cytometric assessment of surface phenotype and intracellular cytokine synthesis was performed. Surface expression of TLR2, TLR4, and FcvarepsilonRI on the monocyte subpopulations was also assessed by means of flow cytometry. RESULTS: TLR2-mediated IL-1beta and TNF-alpha production by either the CD14(dim) or CD14(bright) monocytes was found to be selectively impaired in patients with AD. The most remarkable reduction in TLR2-mediated proinflammatory cytokine production was observed in CD14(dim) monocytes expressing high FcvarepsilonRI levels from patients with AD. This reduction was restored by means of downregulation of their FcvarepsilonRI expression after preculture in the absence of IgE. CONCLUSION: Monocytes, particularly the proinflammatory monocytes, from patients with AD are functionally defective in their capacity to produce proinflammatory cytokines on TLR2 stimuli in part because of the high levels of their FcvarepsilonRI expression. CLINICAL IMPLICATIONS: This selective impairment of monocytes would explain why patients with AD are specifically susceptible to cutaneous staphylococcal and streptococcal and HSV infections.

Hypocholesterolemic mechanism of Chlorella: Chlorella and its indigestible fraction enhance hepatic cholesterol catabolism through up-regulation of cholesterol 7alpha-hydroxylase in rats.

Chlorella powder (CP) has a hypocholesterolemic effect and high bile acid-binding capacity; however, its effects on hepatic cholesterol metabolism are still unclear. In the present study, male Wistar rats were divided into four groups and fed a high sucrose + 10% lard diet (H), an H + 10% CP diet (H+CP), an H + 0.5% cholesterol + 0.25% sodium cholate diet (C), or a C + 10% CP diet (C+CP) for 2 weeks. CP decreased serum and liver cholesterol levels significantly in rats fed C-based diets, but did not affect these parameters in rats fed H-based diets. CP increased the hepatic mRNA level and activity of cholesterol 7alpha-hydroxylase (CYP7A1). CP increased hepatic HMG-CoA reductase (HMGR) activity in the rats fed H-based diets, but not in rats fed C-based diets. CP did not affect hepatic mRNA levels of sterol 27-hydroxylase, HMGR, low-density lipoprotein (LDL) receptor, scavenger receptor class B1, ATP-binding cassette (ABC) A1, ABCG5, or ABCB11. Furthermore, the effect of a 3.08% Chlorella indigestible fraction (CIF, corresponding to 10% CP) on hepatic cholesterol metabolism was determined using the same animal models. CIF also decreased serum and liver cholesterol levels significantly in rats fed C-based diets. CIF increased hepatic CYP7A1 mRNA levels. These results suggest that the hypocholesterolemic effect of CP involves enhancement of cholesterol catabolism through up-regulation of hepatic CYP7A1 expression and that CIF contributes to the hypocholesterolemic effect.

Roles of nuclear receptors in the up-regulation of hepatic cholesterol 7alpha-hydroxylase by cholestyramine in rats.

Nuclear receptors are involved in regulating the expression of cholesterol 7alpha-hydroxylase (CYP7A1), however, their roles in the up-regulation of CYP7A1 by cholestyramine (CSR) are still unclear. In the present study, male Wistar rats were divided into four groups and fed [high sucrose + 10% lard diet] (H), [H + 3% CSR diet] (H + CSR), [H + 0.5% cholesterol + 0.25% sodium cholate diet] (C), or [C + 3% CSR diet] (C + CSR) for 2 weeks. Cholestyramine decreased serum and liver cholesterol levels significantly in rats fed C-based diets, but had no effect on these parameters in rats fed H-based diets. Cholestyramine raised hepatic levels of CYP7A1 mRNA and activity in both groups. The gene expression of hepatic ATP-binding cassettes A1 and G5, regulated by liver X receptor (LXR), were unchanged and down-regulated by cholestyramine, respectively. The mRNA levels of the hepatic ATP-binding cassette B11 and short heterodimer partner (SHP), regulated by farnesoid X receptor (FXR), were not changed by cholestyramine. C-based diets, which contained cholesterol and cholic acid, increased SHP mRNA levels compared to H-based diets. Consequently, in rats fed the C+CSR diet, hepatic FXR was activated by dietary bile acids, but the hepatic CYP7A1 mRNA level was increased 16-fold compared to that in rats fed an H diet. These results suggest that cholestyramine up-regulates the expression of CYP7A1 independently via LXR- or FXR-mediated pathways in rats.

Role of the renal nerves in gamma-aminobutyric acid-induced antihypertensive effect in spontaneously hypertensive rats.

The aim of this study was to clarify the role of the renal sympathetic nerves in the gamma-aminobutyric acid (GABA)-induced hypotensive effect in spontaneously hypertensive rats. Male spontaneously hypertensive rats (SHR/Izm) aged 7 weeks were divided into four groups on the basis of diet (containing 0.05% GABA, or GABA-free control diet) and operation (renal sympathetic-denervated or sham-operated) (n = 10, each). Water intake, urine volume and urinary sodium were, or tended to be, slightly higher, while plasma renin activity was significantly lower in the GABA group than the GABA-free control group. GABA inhibited the development of hypertension in sham-operated spontaneously hypertensive rats but not in renal-denervated spontaneously hypertensive rats. Plasma renin activity was significantly higher in sham-operated spontaneously hypertensive rats fed the control diet than in the other three groups. These results suggest that a reduction in the effects induced by the renal nerves may play an important role in the hypotensive effect induced in spontaneously hypertensive rats by chronic dietary administration of GABA.

T-cell dynamics of inflammatory skin diseases.

The migration of memory T-cells to sites of inflammation is a multistep process controlled by an array of specific receptor-ligand pairs. Chemokines and their receptors represent a central paradigm of the molecular basis of the skin-homing of T-cells. Although CCR4 and CCR10 are both associated with conventionally defined skin-homing T-cells, the association is not necessarily perfect. Interaction between E-selectin and its ligand may represent more specific targets for therapeutic intervention. Although fucosyl transferase?VII is essential for generating E-selectin ligands necessary for T-cell homing to skin, fucosyltransferase-IV, another fucosyltransferase expressed to a significant degree in T-cells, can also generate E-selectin ligands. The induction and upregulation of both enzymes can be co-ordinately regulated depending on their state of activation and differentiation and the cytokine milieu. The dynamic balance between the two enzymes is a major check point for the regulation of skin-homing T-cell differentiation. Polarized T-cells regulate their adhesions on a minute-to-minute basis depending on the cytokine environment. Soluble adhesion molecules found to be increased in chronic inflammatory skin diseases may serve to limit the duration or magnitude of T-cell recruitment. In addition to T-cells migrating from the circulation, T-cells indigenously residing in the tissue itself, such as skin-resident T-cells, would also be responsible for tissue damage. It should also be appreciated that T-cell recruitment to the skin is critical for host defense and that no definitive means exist to distinguish protective regulatory T-cells from pathogenic T-cells.

Effect of a gamma-aminobutyric acid-enriched dairy product on the blood pressure of spontaneously hypertensive and normotensive Wistar-Kyoto rats.

We investigated the blood-pressure-lowering effects of gamma-aminobutyric acid (GABA) and a GABA-enriched fermented milk product (FMG) by low-dose oral administration to spontaneously hypertensive (SHR/Izm) and normotensive Wistar-Kyoto (WKY/Izm) rats. FMG was a non-fat fermented milk product produced by lactic acid bacteria, and the GABA contained in FMG was made from the protein of the milk during fermentation. A single oral dose of GABA or FMG (5 ml/kg; 0.5 mg GABA/kg) significantly (P<0.05) decreased the blood pressure of SHR/Izm from 4 to 8 h after administration, but did not increase that of WKY/Izm rats. The hypotensive activity of GABA was dose-dependent from 0.05 to 5.00 mg/kg in SHR/Izm. During the chronic administration of experimental diets to SHR/Izm, a significantly slower increase in blood pressure with respect to the control group was observed at 1 or 2 weeks after the start of feeding with the GABA or FMG diet respectively (P<0.05) and this difference was maintained throughout the period of feeding. The time profile of blood-pressure change due to administration of FMG was similar to that of GABA. FMG did not inhibit angiotensin 1-converting enzyme. Furthermore, an FMG peptide-containing fraction from reverse-phase chromatography lacked a hypotensive effect in SHR/Izm rats. The present results suggest that low-dose oral GABA has a hypotensive effect in SHR/Izm and that the hypotensive effect of FMG is due to GABA.

In vitro differentiation from naive to mature E-selectin binding CD4 T cells: acquisition of skin-homing properties occurs independently of cutaneous lymphocyte antigen expression.

We previously showed that skin-homing CD4 T cells in peripheral blood can be subdivided into three populations on the basis of the expression pattern of the cutaneous lymphocyte Ag (CLA) and fucosyltransferase VII (FucT-VII): FucT-VII(+)CLA(-), FucT-VII(+)CLA(+), and FucT-VII(-)CLA(+). In view of the known late appearance of CLA during T cell differentiation, T cells programmed to attain skin-homing properties may start to generate E-selectin-binding epitopes at early stages of differentiation before induction of CLA expression. To this end, the in vitro differentiation from naive to CLA(+) memory T cells was followed after activation with anti-CD3 mAb. Here we demonstrate that naive skin-homing CD4 T cell precursors undergo a linear differentiation process from the FucT-VII(+)CLA(-) phenotype to the FucT-VII(+)CLA(+) phenotype and eventually to the FucT-VII(-)CLA(+) phenotype. The appearance of the FucT-VII(+)CLA(-) subset coincided with or could be immediately followed by the generation of E-selectin binding epitopes, and even after E-selectin-binding epitopes were no longer detectable, CLA remained expressed for prolonged periods of time, suggesting that induction of functional E-selectin ligands depends primarily on the expression of FucT-VII, but not CLA. Immunofluorescence and confocal microscopy studies of these T cells confirm that most E-selectin ligands were found independently of CLA expression.

Atypical bullous disease showing features of both erythema multiforme and bullous pemhigoid.

A 78-year-old man presented with multiple, centrifugal erythema, which tended to coalesce, producing polycyclic configurations. The patient developed an annular, narrow blister that was always visible along the margin of the erythema. Histology of a biopsy specimen revealed hydropic degeneration of basal cells, exocytosis of lymphoid cells, and subepidermal blister with necrosis of individual keratinocytes in the blister roof. Direct immunofluorescence studies showed a weak IgG deposition at the basement membrane zone, in a linear fashion, which was confined to the outer side of the blister. Immunoblotting of the patient's serum with human epidermal extract demonstrated circulating antibodies, which reacted to 230 kDa BP antigen 1. These findings suggest that this case is characteristic of both erythema multiforme and bullous pemphigoid and it seems likely that this condition could be a manifestation of epitope spreading, although the exact process in the development of immunological disturbances could not be elucidated.

Direct evidence for interferon-gamma production by effector-memory-type intraepidermal T cells residing at an effector site of immunopathology in fixed drug eruption.

Effector-memory T cells are strategically placed to epithelial tissues to provide frontline immune protection against pathogens. Their detrimental effects, however, have been rarely examined because of difficulty in sampling these T cells in pathological settings. Our previous studies suggested persistence of a similar subset of intraepidermal CD8(+) T cells at high frequencies in the lesions of fixed drug eruption, a localized variant of drug-induced dermatoses. In situ activation of this subset resulting in localized epidermal injury can be traced in the lesions after antigen challenge by paired immunohistochemical staining, reverse transcriptase-polymerase chain reaction in situ, and flow cytometry of dispersed cells. Here we show that effector-memory T cells were greatly enriched in these intraepidermal CD8(+) T cells, but not dermal and circulating counterparts, and that they constitutively express an early activation marker CD69 even before challenge. Surprisingly, a large proportion of these T cells expressed immediate effector function as evidenced by the rapid production of high levels of interferon-gamma in situ with much faster kinetics than their counterparts at the mRNA and protein levels after challenge. This was followed by localized epidermal injury. The intracellular cytokine assay ex vivo shows that the great majority of these dispersed T cells produce interferon-gamma. This study provides the first in situ description of the detrimental effects specifically mediated by effector-memory T cells residing at the effector site of immunopathology.

Involvement of gamma-aminobutyric acid (GABA) B receptors in the hypotensive effect of systemically administered GABA in spontaneously hypertensive rats.

We investigated the effects of intraduodenally (i.d.) administered gamma-aminobutyric acid (GABA) on blood pressure (BP) in anesthetized spontaneously hypertensive rats (SHR) and the mechanism underlying this effect, especially the type of GABA receptor involved in the depressive effect of this amino acid. GABA (0.3 to 300 mg/kg, i.d.) caused a dose-related decrease in the BP of 9.20 +/- 3.96 to 35.0 +/- 5.34 mmHg (mean +/- S.E.M.) that lasted for 30 to 50 min. The minimum effective i.d. dose of GABA was 0.3 to 1.0 mg/kg. Results pertaining to the mechanism underlying the GABA-induced effects on BP were as follows: a) GABA did not alter the BP-related effects of exogenous noradrenaline and acetylcholine; b) pretreatment with hexamethonium decreased the GABA-induced fall in BP, and GABA tended to reduce the pressor response associated with injection of dimethyl phenylpiperazinium; and c) pretreatment with 2-hydroxysaclofen markedly reduced the GABA-induced drop in BP, whereas pretreatment with bicuculline did not. In conclusion, in SHR, low-dose (0.3 to 1.0 mg/kg, i.d.) GABA had a hypotensive effect, which may result from attenuation of sympathetic transmission through the activation of GABA(B) receptors at presynaptic or ganglionic sites.