Primo vascular system in human umbilical cord and placenta.

The primo vascular system (PVS) has been observed in various animals such as mice, rats, rabbits, dogs, swine, and cow, but not in humans. In this work, we report on the observation of a human PVS on both the epithelial fascia and inside the blood vessels of the umbilical cord (UC). The main morphological characteristics of the primo vessels (PVs) and primo nodes (PNs) from the human UC were in agreement with those of the PVS in various animal organs, including the thicknesses and the transparency of the PVs, the sizes of the PNs, the broken-line arrangement of the rod-shaped nuclei, the sparse distribution of nuclei, and the presence of hollow lumens in the central inner parts of the PNs. It was rather surprising that the human PV was not thicker than the PVs from small animals. The difference between the PVS and blood/lymph vessels was confirmed using immunofluorescence staining of von Willebrand factor, CD31, LYVE-1, and D2-40. The positive expression of the PVS to proliferating cell nuclear antigen, a cell-proliferation marker, was consistent with the recent finding of very small embryonic-like stem cells in the PVS of mice.

Mycorrhizal colonisation and P-supplement effects on N uptake and N assimilation in perennial ryegrass under well-watered and drought-stressed conditions.

To compare the effect of arbuscular mycorrhiza (AM) and P-supplement on N uptake and N assimilation under well-watered or drought-stressed conditions, Glomus intraradices-colonised, P-supplemented non-mycorrhizal (P) and non-mycorrhizal (control) plants of Lolium perenne were exposed to 12 days of water treatment. Leaf water potential (Ψ (w)), photosynthetic ability, and N and P nutritional status were measured at the beginning (day 0) and end (day 12) of water treatment. N absorption, amino acid and protein synthesis were quantified using the isotopic tracer (15)N at day 12. Under well-watered conditions, growth response and physiological parameters were similar in AM and P plants, as compared to controls. Drought (10% water) significantly decreased these parameters in all three treatments. As compared to control plants, the negative impact of water deficit on the Ψ (w), photosynthesis, biomass, and N and P content was highly alleviated in AM plants, while only slightly improved or remained the same level in P plants. The effect of AM symbiosis on N absorption and N assimilation was greater than that of the P supplement under well-watered and drought-stressed conditions, and this effect was highly enhanced under drought-stressed conditions. At terminal drought stress on day 12, the effect of AM colonisation on de novo synthesis of amino acids and proteins was 4.4- and 4.8-fold higher than that of the P supplement. These results indicate that the AM symbiosis plays an integrative role in N nutrition by alleviating the negative impacts of drought on N or P uptake and N assimilation, whereas the efficiency of a direct P supplement is very limited under drought-stressed conditions.

Epigallocatechin-3-gallate inhibits paracrine and autocrine hepatocyte growth factor/scatter factor-induced tumor cell migration and invasion.

Aberrant activation of hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, Met, is involved in the development and progression of many human cancers. In the cell-based screening assay, (-)epigallocatechin-3-gallate (EGCG) inhibited HGF/SF-Met signaling as indicated by its inhibitory activity on HGF/SF-induced cell scattering and uPA activation (IC50=15.8 microgram/ml). Further analysis revealed that EGCG at low doses specifically inhibited HGF/SF-induced tyrosine phosphorylation of Met but not epidermal growth factor (EGF)-induced phosphorylation of EGF receptor (EGFR). On the other hand, high-dose EGCG decreased both Met and EGFR proteins. We also found that EGCG did not act on the intracellular portion of Met receptor tyrosine kinase, i.e., it inhibited InlB-dependent activation of Met but not NGF-induced activation of Trk-Met hybrid receptor. This inhibition decreased HGF-induced migration and invasion by parental or HGF/SF-transfected B16F10 melanoma cells in vitro in either a paracrine or autocrine manner. Furthermore, EGCG inhibited the invasion/metastasis of HGF/SF-transfected B16F10 melanoma cells in mice. Our data suggest the possible use of EGCG in human cancers associated with dysregulated paracrine or autocrine HGF/SF-Met signaling.

The herbal extract HMC05 inhibits neointima formation in balloon-injured rat carotid arteries: possible therapeutic implications of HMC05.

AIM OF THE STUDY: In a previous study, HMC05, a water extract from eight medicinal herbs was demonstrated to possess anti-inflammatory effects in murine macrophages and anti-atherosclerotic effects in apoE(-/-) mice. HSP27 expression was shown to be decreased in advanced atherosclerotic plaques of human carotid arteries. In the present study, the role of HMC05 in the prevention of restenosis and the possible mechanisms involved in the decrease of neointima formation were investigated using in vivo balloon injury rat model and in vitro biochemical assays. MATERIALS AND METHODS: A rat carotid artery balloon injury restenosis model was used. Different doses of HMC05 were administered to the rats by tube feeding, starting from four days before surgery and continuing twice per week for two weeks after carotid injury. Injured carotid arteries isolated from rats were embedded in paraffin block and tissue sections were stained with H&E to assess neointima formation. Mechanism by HMC05 that are involved in smooth muscle cell proliferation and migration was assessed by western blot assay, immunohistochemistry and confocal analysis. RESULTS: There was no significant difference in the medial area between the control and HMC05-treated groups. However, neointima formation was significantly inhibited in the HMC05-treated group, resulting in 47-fold lower intima to media ratios in rats treated with 25 mg/kg/day HMC05 as compared to the control. Surprisingly, monocytes infiltration in the neointima area was almost completely blocked by HMC05 administration. When rat vascular SMCs were treated with HMC05, the proliferation and migration of smooth muscle cells was dramatically inhibited in a dye uptake assay and in a scratch model in a culture dish, respectively. HMC05 dose-dependently inhibited PDGF-mediated MAPK and AKT activation. However, HMC05 did not affect PDGF-mediated HSP27 phosphorylation but it induced HSP27 overexpression and phosphorylation. In addition, medial SMCs in the arterial wall of rats treated with HMC05 showed a significant increase in HSP27 expression compared with that of the control rats. CONCLUSIONS: HMC05, a strong anti-inflammatory reagent, might use HSP27 as an effector molecule in SMCs to reduce neointimal hyperplasia by inhibiting PDGF-mediated MAPK and AKT activation. HMC05 could be a useful drug candidate for the prevention of restenosis after balloon injury of the arteries.

Therapeutic effect of hyperbaric oxygen in psoriasis vulgaris: two case reports and a review of the literature.

INTRODUCTION: Psoriasis is an inflammatory and immunological cutaneous disease. The high morbidity in patients with psoriasis results from severe clinical manifestations and/or adverse effects of treatment. The Undersea and Hyperbaric Medical Society and Federal Medicare and Medicaid Services have approved the use of hyperbaric oxygen (HBO(2)) for more than 15 indications, including wound healing, infections and late effects of radiation, which are largely unresponsive to conventional treatments. Accumulated data show that HBO(2) has anti-inflammatory effects and other positive influences on the immune system, making it a rational treatment in the management of psoriasis plaques and arthritis. CASE PRESENTATION: We present the cases of two patients with long histories of psoriasis vulgarus who exhibited marked improvement with use of HBO(2.) The first patient was 40 years old and had pustular psoriasis and psoriatic arthritis. He was treated with six sessions of HBO(2) (at 2.8 atmospheres of pressure for 60 minutes), which successfully controlled his symptoms. At the 18-month post-treatment follow up, the patient exhibited complete remission of psoriasis and marked improvement in psoriatic arthritis without medication. The second patient was 55 years old with extensive psoriatic lesions, and exhibited marked improvement within 15 sessions of HBO(2). No adverse effects of HBO(2) were identified. CONCLUSIONS: HBO(2) may possess potential therapeutic efficacy in the management of psoriasis. We outline the pathogenesis of psoriasis and the selective anti-inflammatory and immunosuppressive effects of HBO(2). We hope that this will provide a basis for elucidating the mechanisms of action and consequently pave the way for further controlled studies.

Ultra-low microcurrent in the management of diabetes mellitus, hypertension and chronic wounds: report of twelve cases and discussion of mechanism of action.

Oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus and cardiovascular diseases including hypertension. The low levels of antioxidants accompanied by raised levels of markers of free radical damage play a major role in delaying wound healing. Ultra-low microcurrent presumably has an antioxidant effect, and it was shown to accelerate wound healing. The purpose of the study is to investigate the efficacy of ultra-low microcurrent delivered by the Electro Pressure Regeneration Therapy (EPRT) device (EPRT Technologies-USA, Simi Valley, CA) in the management of diabetes, hypertension and chronic wounds. The EPRT device is an electrical device that sends a pulsating stream of electrons in a relatively low concentration throughout the body. The device is noninvasive and delivers electrical currents that mimic the endogenous electric energy of the human body. It is a rechargeable battery-operated device that delivers a direct current (maximum of 3 milliAmperes) of one polarity for 11.5 minutes, which then switched to the opposite polarity for another 11.5 minutes. The resulting cycle time is approximately 23min or 0.000732 Hz and delivers a square wave bipolar current with a voltage ranging from 5V up to a maximum of 40 V. The device produces a current range of 3 mA down to 100 nA. Twelve patients with long standing diabetes, hypertension and unhealed wounds were treated with EPRT. The patients were treated approximately for 3.5 h/day/5 days a week. Assessment of ulcer was based on scale used by National Pressure Ulcer Advisory Panel Consensus Development Conference. Patients were followed-up with daily measurement of blood pressure and blood glucose level, and their requirement for medications was recorded. Treatment continued from 2-4 months according to their response. Results showed that diabetes mellitus and hypertension were well controlled after using this device, and their wounds were markedly healed (30-100%). The patients either reduced their medication or completely stopped after the course of treatment. No side effects were reported. The mechanism of action was discussed.

HMCO5, herbal extract, inhibits NF-kappaB expression in lipopolysaccharide treated macrophages and reduces atherosclerotic lesions in cholesterol fed mice.

HMCO5 is a herbal extract which comprises of eight different herbs. We studied whether this extract has anti-atherosclerotic effects. In lipopolysaccharide (LPS) stimulated RAW264.7 cells, HMCO5 inhibited NF-kappaB activation as well as iNOS promoter activity, inhibited the secretion of TNF-alpha and IL-1beta, and directly inhibited the intracellular accumulation of reactive oxygen species. ApoE knock-out mice fed a high-fat high-cholesterol diet with HMCO5 for 10 weeks showed a significant reduction in atherosclerotic lesions. A notable finding was the preservation of the smooth muscle cell layer in the media of aorta in the HMCO5 co-treated mice. HMCO5 treated mice did not show significant decrease in serum level of cholesterol. These results suggest that HMCO5 has anti-atherosclerotic effects which in part may be attributable to the inhibition of production of NF-kappaB dependent pro-inflammatory cytokines.

Wogonin suppresses TARC expression induced by mite antigen via heme oxygenase 1 in human keratinocytes. Suppressive effect of wogonin on mite antigen-induced TARC expression.

BACKGROUND: Mite antigen, extract from Dermatophagoides farinae in house dust, is a well-known causative agent of atopy or allergic diseases, which involves many inflammatory cytokines/chemokines expression. Heme oxygenase 1 (HO1) has recently emerged as an important cytoprotective enzyme against oxidative stress and inflammatory responses in many cell types. OBJECTIVE: The aim of this study was to investigate the possible mechanism by which wogonin, a natural product isolated from Scutellaria baicalensis, inhibited the mite antigen-induced chemokine expression in human keratinocytes, HaCaT cells. METHODS: The level of chemokine expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) and signaling study was performed by Western blot analysis. RESULTS: The mite antigen-induced thymus- and activation-regulated chemokine (TARC/CCL17) expression in a dose-dependent manner via extracellular signal-regulated kinase (ERK) activation. However, it did not affect the expression of other chemokines including macrophage-derived chemokine (MDC/CCL22), RANTES, and IL-8. Interestingly, wogonin significantly suppressed the mite antigen-induced TARC expression via the induction of HO1. This suppression was completely restored by HO1 siRNA, suggesting a direct role of HO1 for the suppressive effect. Furthermore, exogenous CO, but not other end products of HO1 activity, also suppressed the mite antigen-induced TARC expression. CONCLUSION: Wogonin induces HO1 expression, which in turn HO1 and/or CO suppresses TARC expression induced by mite antigen in human HaCaT cells.

Ultra-low microcurrent therapy: a novel approach for treatment of chronic resistant wounds.

This study was undertaken to investigate the efficacy of ultra-low microcurrent delivered by the Electro Pressure Regeneration Therapy (EPRT) device for the management of chronic wounds. In this study, 23 patients with chronic skin ulcers and 2 with abdominal dehiscence that was present for an average of 16.5 mo, who were not responsive to standard conservative treatment in a hospital setting, were treated with the EPRT device. Wounds were treated with direct current (maximum of 3 mA) of 1 polarity for 11.5 min and then with a current of the opposite polarity for another 11.5 min. Treatment was applied through ultra-low microcurrents (in the mA to nA range) conducted through special wraps applied above and below the wound. The results revealed that 34.8% of cases achieved complete wound healing after an average of 45.6 h of treatment, and 39.1% achieved >or=50% healing after an average of 39.7 h of treatment. Several patients achieved significant results after 1 to 2 treatments. The EPRT device not only accelerated healing but also appeared to negate the effect of a person's age on wound healing.

Influences of hyperbaric oxygen on blood pressure, heart rate and blood glucose levels in patients with diabetes mellitus and hypertension.

BACKGROUND: We investigated the influences of hyperbaric oxygen (HBO(2)) on systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR) and blood glucose level (BGL). METHODS: Forty one patients with hypertension (HTN), diabetes mellitus (DM), HTN and DM and/or no HTN or DM underwent HBO(2) sessions (15-40 sessions for each patient). SBP, DBP, HR and BGL (for diabetics) were recorded before and after each session. RESULTS: HBO(2) caused significant elevation in SBP (11%) and DBP (12%) and a decrease in HR (18%) (p <0.001). Patients with DM and HTN showed higher elevation in SBP and DBP. HBO(2) lowered BGL by 23% (p <0.001). When basal BGL was in the range of 120-170 mg/dl, it dropped to <100 mg/dl in 31/60 treatment sessions (52%). When basal BGL was <120 mg/dl it dropped to <70 mg/dl in 8/34 sessions. There was a possibility of lowered BGL when basal BGL was <170 mg/dl and a marked reduction in BGL occurred when basal BGL was <120 mg/dl. HBO(2) caused a marked elevation in SBP and DBP when basal SBP was >140 mmHg. Critical elevation was obtained when SBP was >160 mmHg. The use of beta blockers caused significant elevation of blood pressure while reducing HR. CONCLUSIONS: HBO(2) causes elevation of blood pressure and lowering of HR and BGL, which were augmented in the presence of HTN, DM, or beta blocker. The use of beta blockers for the management of HTN should be avoided during HBO(2) therapy.

Polyozellin inhibits nitric oxide production by down-regulating LPS-induced activity of NF-kappaB and SAPK/JNK in RAW 264.7 cells.

Polyozellin, isolated from Polyozellus multiplex (Thelephoraceae), was investigated for its anti-inflammatory activity in the murine macrophage cell line RAW 264.7. Polyozellin inhibited both lipopolysaccharide (LPS)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) gene expression in a dose-dependent manner. The effects of polyozellin on the activation of nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein (MAP) kinases in these cells were studied in order to elucidate the underlying mechanism. Polyozellin suppressed the activation of both LPS-induced NF-kappaB and the stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), but had no effect on the extracellular signal-regulated kinase (ERK) or p38. These data suggest that polyozellin suppresses iNOS expression by inhibiting the activation of NF-kappaB and SAPK/JNK, leading to the inhibition of NO production.

A synthetic peptidoglycan fragment as a competitive inhibitor of the melanization cascade.

Melanin synthesis is essential for defense and development but must be tightly controlled because systemic hyperactivation of the prophenoloxidase and excessive melanin synthesis are deleterious to the hosts. The melanization cascade of the arthropods can be activated by bacterial lysine-peptidoglycan (PGN), diaminopimelic acid (DAP)-PGN, or fungal beta-1,3-glucan. The molecular mechanism of how DAP- or Lys-PGN induces melanin synthesis and which molecules are involved in distinguishing these PGNs are not known. The identification of PGN derivatives that can work as inhibitors of the melanization cascade and the characterization of PGN recognition molecules will provide important information to clarify how the melanization is regulated and controlled. Here, we report that a novel synthetic Lys-PGN fragment ((GlcNAc-MurNAc-L-Ala-D-isoGln-L-Lys-D-Ala)2, T-4P2) functions as a competitive inhibitor of the natural PGN-induced melanization reaction. By using a T-4P2-coupled column, we purified the Tenebrio molitor PGN recognition protein (Tm-PGRP) without causing activation of the prophenoloxidase. The purified Tm-PGRP recognized both Lys- and DAP-PGN. In vitro reconstitution experiments showed that Tm-PGRP functions as a common recognition molecule of Lys- and DAP-PGN-dependent melanization cascades.

Hyperbaric oxygen and malignancies: a potential role in radiotherapy, chemotherapy, tumor surgery and phototherapy.

Over the past 40 years, hyperbaric oxygen (HBO2) therapy has been recommended and used in a wide variety of medical conditions. In the 1950s, HBO2 was first used as a treatment, in addition to radiation, for head and neck cancers and cervical cancer. Many studies have been conducted to investigate possible therapeutic effects HBO2 as part of cancer management. Evidences showed that HBO2 improved tumor oxygenation, and treatment with HBO2 during irradiation has been shown to improve the radiation response of many solid tumors. It was used for delayed radiation injuries for soft tissue and bony injuries, for symptomatic radiation reactions of the urinary bladder and the bowel, for laryngeal radionecrosis, for radiation-induced optic neuropathy, for radiation-induced proctitis and for radiation-induced necrosis of the brain. HBO2 also increases sensitivity to chemotherapy. A significant improvement in tumor response was obtained when photodynamic therapy (PDT) was delivered during hyperoxygenation. These studies were extensively reviewed and rational scientific basis for further investigations was discussed. The possibility of combining HBO2, PDT and photosensitizers to overcome primary and secondary carcinoma deserve extensive laboratory and clinical research works. HBO2 is a relatively benign with few contraindications, even for active cancer patients.

A zymogen form of masquerade-like serine proteinase homologue is cleaved during pro-phenoloxidase activation by Ca2+ in coleopteran and Tenebrio molitor larvae.

To elucidate the biochemical activation mechanism of the insect pro-phenoloxidase (pro-PO) system, we purified a 45-kDa protein to homogeneity from the hemolymph of Tenebrio molitor (mealworm) larvae, and cloned its cDNA. The overall structure of the 45-kDa protein is similar to Drosophila masquerade serine proteinase homologue, which is an essential component in Drosophila muscle development. This Tenebrio masquerade-like serine proteinase homologue (Tm-mas) contains a trypsin-like serine proteinase domain in the C-terminal region, except for the substitution of Ser to Gly at the active site triad, and a disulfide-knotted domain at the amino-terminal region. When the purified 45-kDa Tm-mas was incubated with CM-Toyopearl eluate solution containing pro-PO and other pro-PO activating factors, the resulting phenoloxidase (PO) activity was shown to be independent of Ca2+. This suggests that the purified 45-kDa Tm-mas is an activated form of pro-PO activating factor. The55-kDa zymogen form of Tm-mas was detected in the hemolymph when PO activity was not evident. However, when Tenebrio hemolymph was incubated with Ca2+, a 79-kDa Tenebrio pro-PO and the 55-kDa zymogen Tm-mas converted to 76-kDa PO and 45-kDa Tm-mas, respectively, with detectable PO activity. Furthermore, when Tenebrio hemolymph was incubated with Ca2+ and beta-1,3-glucan, the conversion of pro-PO to PO and the 55-kDa zymogen Tm-mas to the 45-kDa protein, was faster than in the presence of Ca2+ only. These results suggest that the cleavage of the 55-kDa zymogen of Tm-mas by a limited proteolysis is necessary for PO activity, and the Tm-mas is a pro-PO activating cofactor.