Hypoxia improves hair inductivity of dermal papilla cells via nuclear NADPH oxidase 4-mediated reactive oxygen species generation'.

BACKGROUND: Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis. Therefore, tremendous efforts have been made to promote DPC hair inductivity. OBJECTIVES: The aim of this study was to investigate the mitogenic and hair inductive effects of hypoxia on DPCs and examine the underlying mechanism of hypoxia-induced stimulation of DPCs. METHODS: DPCs' hair inductivity was examined under normoxia (20% O2 ) and hypoxia (2% O2 ). RESULTS: Hypoxia significantly increased the proliferation and delayed senescence of DPCs via Akt phosphorylation and downstream pathways. Hypoxia upregulated growth factor secretion of DPCs through the mitogen-activated protein kinase pathway. Hypoxia-preconditioned DPCs induced the telogen-to-anagen transition in C3 H mice, and also enhanced hair neogenesis in a hair reconstitution assay. Injected green fluorescent protein-labelled DPCs migrated to the outer root sheath of the hair follicle, and hypoxia-preconditioning increased survival and migration of DPCs in vivo. Conditioned medium obtained from hypoxia increased the hair length of mouse vibrissa follicles via upregulation of alkaline phosphatase, vascular endothelial growth factor, and glial cell line-derived neurotrophic factor. We examined the mechanism of this hypoxia-induced stimulation, and found that reactive oxygen species (ROS) play a key role. For example, inhibition of ROS generation by N-acetylcysteine or diphenyleneiodonium treatment attenuated DPCs' hypoxia-induced stimulation, but treatment with ROS donors induced mitogenic effects and anagen transition. NADPH oxidase 4 is highly expressed in the DPC nuclear region, and NOX4 knockout by CRISPR-Cas9 attenuated the hypoxia-induced stimulation of DPCs. CONCLUSIONS: Our results suggest that DPC culture under hypoxia has great advantages over normoxia, and is a novel solution for producing DPCs for cell therapy. What's already known about this topic? Dermal papilla cells (DPCs) play a key role in hair regeneration and morphogenesis, but they are difficult to isolate and expand for use in cell therapy. Tremendous efforts have been made to increase proliferation of DPCs and promote their hair formation ability. What does this study add? Hypoxia (2% O2 ) culture of DPCs increases proliferation, delays senescence and enhances hair inductivity of DPCs. Reactive oxygen species play a key role in hypoxia-induced stimulation of DPC. What is the translational message? Preconditioning DPCs under hypoxia improves their hair regenerative potential, and is a novel solution for producing DPCs for cell therapy to treat hair loss.

Antimicrobial activities of an oxygenated cyclohexanone derivative isolated from Amphirosellinia nigrospora JS-1675 against various plant pathogenic bacteria and fungi.

AIMS: To evaluate the antimicrobial activities of an active compound isolated from the culture broth of Amphirosellinia nigrospora JS-1675 against various plant pathogenic bacteria and fungi. METHODS AND RESULTS: While screening for bioactive secondary metabolites from endophytic fungi, we found that A. nigrospora JS-1675 showed strong in vitro antibacterial activity against Ralstonia solanacearum. One compound (1) was isolated and identified as (4S, 5S, 6S)-5,6-epoxy-4-hydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one. Growth of most of the tested phytopathogenic bacteria was inhibited by compound 1 and the ethyl acetate (EtOAc) layer except Pseudomonas syringae pv. lachrymans. Compound 1 also inhibited the mycelial growth of several plant pathogenic fungi. Both compound 1 and the EtOAc layer reduced bacterial leaf spot disease in detached peach leaves. They also suppressed the development of bacterial wilt on tomato seedlings quite effectively. CONCLUSIONS: Amphirosellinia nigrospora JS-1675 showed antimicrobial activity against plant pathogenic bacteria and fungi by producing compound 1. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the occurrence of compound 1 in A. nigrospora JS-1675 and its efficacy against plant pathogenic bacteria and fungi. Their strong disease control efficacy against tomato bacterial wilt suggests that this fungus can be used as a microbial bactericide.

Renal failure and progressive pancytopenia.

Hyperparathyroidism may be a precipitating factor to the development of myelofibrosis; however, this is extremely rare with only a few documented case reports of myelofibrosis caused by secondary hyperparathyroidism. We describe a case of a 24-year-old female who had a failed live donor renal transplant and secondary hyperparathyroidism. While on haemodialysis she became increasingly pancytopenic despite erythropoietin injections and adequate iron, vitamin B12 and folate replacement. Her secondary hyperparathyroidism evolved to tertiary hyperparathyroidism despite vitamin D supplementation and phosphate binders. In order to determine the cause of her pancytopenia, a bone marrow biopsy was performed that confirmed myelofibrosis due to her secondary hyperparathyroidism. Following a successful parathyroidectomy in a tertiary hospital, her pancytopenia resolved and she is now awaiting a second transplant.

Antibacterial activities of penicillic acid isolated from Aspergillus persii against various plant pathogenic bacteria.

UNLABELLED: The emergence of pathogenic bacterial strains resistant to agrochemicals and the increasing demand for organic foods have led to the discovery of new antibacterial metabolites that can be used either directly or as a lead molecule for development of synthetic bactericides. During the screening of antibacterial fungal cultures, we found that one fungal strain, Aspergillus persii EML-HPB1-11, showed strong in vitro antibacterial activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibitory concentration (MIC) of 10% of fermentation broth filtrate. The active compound was identified as penicillic acid (PA: 3-methoxy-5-methyl-4-oxo-2,5-hexadienoic acid) by mass and NMR spectroscopy. The in vitro antibacterial activity of PA was tested against 12 phytopathogenic bacteria. All of the bacterial pathogens tested were highly inhibited by PA with MIC values of 12·3-111·1 µg ml(-1) . It also effectively suppressed the development of bacterial spot disease in detached peach leaves, showing control values of 82·4 and 94·1% at concentrations of 111·1 and 333·3 µg ml(-1) respectively. This is the first report on the production of PA by A. persii. This study suggests that PA can be used as a lead molecule for development of synthetic bactericides for control of various plant diseases. SIGNIFICANCE AND IMPACT OF THE STUDY: Penicillic acid (PA) produced by the seed-borne fungus Aspergillus persii EML-HPB1-11 showed antibacterial activity against various plant pathogenic bacteria. The compound effectively inhibited the growth of 12 plant pathogenic bacteria and successfully controlled bacterial spot disease on peach leaf. These results suggest that PA can be used as a lead molecule for development of synthetic agrochemicals to control plant bacterial diseases.

Contrast echocardiography to assess left ventricular volume and function in Beagle dogs: comparison with 3-Tesla dual source parallel cardiac magnetic resonance imaging.

This study was performed to evaluate the effect and feasibility of contrast echocardiography (CE) compared with unenhanced echocardiography (UE) and cardiac magnetic resonance imaging (CMRI) to assess left ventricular (LV) volume and function, including end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), and ejection fraction (EF) in six healthy Beagles. When the dogs were conscious, LV measurements using CE were significantly higher than those obtained using UE, except for EF, and were similar to the values obtained using CMRI. Additionally, EDV, SV, and EF obtained using UE from anesthetized dogs were significantly lower than those obtained using CE or CMRI. Measurements of EDV, SV and EF using CE were not significantly different from the corresponding measurements obtained using CMRI (31.13±2.18 vs. 32.88±1.17 mL, 18.41±1.25 vs. 17.92±0.96 mL, 59.29±2.29% vs. 53.33±1.69%, respectively). Inter-observer agreements for UE (0.74±0.05) were lower than those for CE (0.80±0.04) and CMRI (0.92±0.03). In conclusion, LV function was assessed reproducibly using CE, and the measurements obtained were consistent with reference standard measurements obtained using CMRI. Measurements made using CE agreed more closely with CMRI than those made using UE.

Effects of honeybee venom supplementation in drinking water on growth performance of broiler chickens.

The effects of water supplementation of bee venom (BV) on performance, antioxidant activity, and liver function in Arbor Acres broiler chickens were investigated. Hence, 3 experimental treatment groups (control, 0.5 mg/L of BV, and 1 mg/L of BV) were allocated to 3 replicates of 5,000 one-day-old chicks each. The control group was kept on tap water, whereas the other 2 groups were supplied water supplemented with 0.5 and 1 mg of BV, respectively, per liter of drinking water. Broilers were provided ad libitum access to feed for the experimental period of 1 to 28 d of age. Supplementing drinking water with BV significantly increased BW gain at 28 d of age (P < 0.05). The average daily weight gain from d 1 to 28 was increased for birds supplemented with BV compared with control birds. The increase in BW gain was more pronounced with supplementation of 1 mg/L of BV compared with 0.5 mg/L of BV. An improved feed intake was noted in groups supplemented with BV as compared with control chicks. Liver function enzymes, aspartate aminotransferase, and alanine aminotransferase activities including total cholesterol, total protein, albumin, and globulin were not changed by BV supplementation. Tap water supplementation of BV did not alter the number of leukocytes, erythrocytes, heterophils, and lymphocytes. However, the antioxidative activities estimated as a superoxide dismutase-like activity of broiler chicks supplemented with BV was significantly increased (P < 0.05) in comparison with those without BV supplementation. These data indicate a possibility of better broiler performance through BV supplementation under conditions of severe stressful challenges the newly born chicks encounter.

Computational modelling of blood flow through curved stenosed arteries.

A computational model of three-dimensional blood flow in curved arteries with elliptic stenosis was developed. Two groups of models, (a) different angles of curvature and (b) degrees of stenosis, have been studied under typical conditions for stenosed coronary artery. Useful information on the haemodynamics has been obtained. Results of pressure drop show that the presence of the curvature augments the increased flow resistance due to stenotic lesions. The study also demonstrates the significant presence of secondary flow in a curved artery. In addition, the results have shown that the secondary flow in a curved artery brings about elevated shear stress on the vessel wall. These results indicated that both curvature and stenosis should be considered together by cardiologists to assess or quantify the severity of the stenosis. This study employed a powerful computer-aided design (CAD) package to construct the model and a commercial computational fluid dynamics (CFD) code for the analysis of blood flow in stenosed arteries. The long-term application of this form of research promises to be an effective tool for gaining insights into the pathology of arterial diseases.

The effects of Pva/chitosan/fibroin (PCF)-blended spongy sheets on wound healing in rats.

The effects of poly vinyl alcohol (PVA)/Chitosan/Fibroin (PCF)-blended sponge on wound healing in rats were investigated. We excised the skin of a rat, including the dermis, approximately 2 x 2 cm in size. The wound was covered with PCF-blended spongy sheets. The spongy sheets absorbed the exudate, and gained flexibility and softness. Histopathological inspection of the wound 12 d later showed an increase of vascular ingrowth and the absence of inflammatory cells. Regeneration of the skin around the wound was faster than that of the control. We also tested wound healing effects of PVA, Chitosan and Fibroin, alone or in various combinations. Wound healing was accelerated in the order of PCF-blended sponge>Chitosan/Fibroin (CF)-blended sponge>Fibroin (F) sponge > PVA/Chitosan-blended sponge (PC) > Chitosan (C) sponge.

Synthesis and antimicrobial activity of novel 3-[(aminopyrimidiniumyl)thio]methyl cephalosporins.

A series of novel cephalosporin compounds which have 3-[(aminopyrimidiniumyl)thio]methyl substituents was synthesized. They show high antimicrobial activity against various bacterial species including Pseudomonas aeruginosa. Structure-activity relationships with various thiopyrimidines, thiopyrimidiniums, bicyclic thiotriazolopyrimidiniums, and bicyclic thioimidazolopyrimidiniums as 3'-substituents were also studied; cephalosporins with quarternized pyrimidinium moieties have better antimicrobial activities than neuteral pyrimidine cephalosporins, and stabilization of the positive charge on the pyrimidinium moieties is essential for better activity. According to semiempirical PM3 calculations, amino and alkylthio substituents on the pyrimidinium rings play a major role in charge stabilization and delocalization.

Structural analysis of highly oriented poly(p-phenylene-terephthalamide) by 15N solid-state nuclear magnetic resonance.

The structure of an aromatic polyamide, poly(p-phenylene-terephthalamide) (PPTA), was studied in the solid state using 15N nuclear magnetic resonance (NMR) spectroscopy. Spectra of uniaxially aligned molecules placed with the axis of alignment both parallel with and perpendicular to the applied magnetic field were analyzed to yield the orientations of specific molecular bonds with respect to the fiber axis. The 15N chemical shift tensor was characterized by simulating powder pattern spectra of both PPTA and a model compound, benzanilide. Chemical shift and dipolar coupled chemical shift line shapes were calculated through Euler angle transformations from the principal axis system (PAS) reference frame to the fiber axis system (FAS) frame. The orientations of NH and NC' bonds in PPTA are determined as well as the orientational distribution of the PPTA fiber axis. The structural parameters determined for PPTA are compared with those obtained by X-ray diffraction.

The rate of sulfur hexafluoride escape from a plastic syringe.

Sulfur hexafluoride (SF6) gas is widely used for internal tamponade during retinal reattachment surgery and is commonly injected into the eye from a 10-mL plastic syringe. The rate of diffusion of SF6 out of a plastic syringe has not bee studied. We measured the percentage of SF6 gas in a 10-mL plastic syringe by gas chromatography, confirmed by infrared spectrometry. Measurements were obtained immediately after aspiration, and at 30 s and 10, 15, 60, 90, and 120 minutes, and 18 hours. A marked decrease in SF6 concentration, from 97% at 30 s to 76% at 60 minutes and 2% at 18 hours, was noted. The results were highly reproducible. Sulfur hexafluoride gas should be injected into the patient's eye as soon as possible after aspiration from the tank to ensure accurate concentrations.

Photocoagulation to prevent retinal detachment in acute retinal necrosis.

Retinal detachment (RD) occurs in more than 50% of eyes with acute retinal necrosis (ARN) and is the leading cause of visual loss in this syndrome. In order to decrease the incidence of RD in ARN, the authors treated 12 eyes of 10 patients with prophylactic laser photocoagulation. Retinal detachment occurred in two eyes (17%). Over the same time period, seven eyes with ARN did not receive prophylactic laser treatment, most often because of dense vitreous debris, with a 67% rate of RD. Prophylactic photocoagulation treatment should be considered in the management of patients with ARN.

Interphotoreceptor retinoid-binding protein levels in subretinal fluid from rhegmatogenous retinal detachment and retinopathy of prematurity.

Interphotoreceptor retinoid-binding protein (IRBP) has been detected qualitatively in subretinal fluid associated with rhegmatogenous retinal detachments. We have used a quantitative enzyme-linked immunosorbent assay to determine the IRBP concentrations in 24 fluid specimens collected at retinal reattachment surgery from 22 patients, and from 17 patients who underwent surgery for retinopathy of prematurity with traction detachment. We observed a wide range of concentrations (1 to 176 micrograms of IRBP per milligram of protein). Higher concentrations were found only in fluid from the more recent rhegmatogenous detachments. The lowest concentrations were detected among fluids drained from under rhegmatogenous detachments ranging from five days' to one year's duration. The IRBP concentration in subretinal fluid showed no correlation with visual outcome and may be influenced by IRBP turnover in the vitreous cavity. Most strikingly, none of the 17 samples from patients with retinopathy of prematurity contained detectable amounts of IRBP.

Retinal pigment epithelial tears. Patterns and prognosis.

Increasing experience with the diagnosis of retinal pigment epithelial (RPE) tears has led to expanded recognition and understanding of this clinical entity. The authors report 18 RPE tears followed for an average of 28 months; 16 were associated with age-related macular degeneration and 2 with presumed ocular histoplasmosis syndrome. Retinal pigment epithelial dehiscences fell into four categories: nine spontaneous tears associated with choroidal neovascularization, one tear associated with an RPE detachment without choroidal neovascularization, four iatrogenic tears occurring at krypton treatment of choroidal neovascularization, and four iatrogenic tears developing weeks to months after laser treatment of choroidal neovascularization. Eight patients had a final visual acuity of 20/100 or better, four were 20/200, and six were 20/400 or worse. Photocoagulation, particularly with the use of krypton red laser, may be modified on the basis of possible RPE tear formation. Heightened awareness of the possibility of inducing pigment epithelial rips should improve diagnosis and management of these cases.

Silicone oil in the treatment of complicated retinal detachments.

Silicone oil is now used with increasing frequency to treat cases of complicated retinal detachment (RD). The authors report their results using silicone oil in eyes with RD resulting from proliferative diabetic retinopathy (PDR) or proliferative vitreoretinopathy (PVR). The retina was completely reattached at last examination in 16 (70%) of 23 eyes with PDR and 20 (67%) of 30 eyes with PVR. Final vision of 5/200 or better was obtained in 5 eyes with PDR (22%) and 16 eyes with PVR (53%) (P = 0.0016). Reproliferation of epiretinal membranes occurred in 26% of diabetics and 23% of eyes with PVR. Silicone oil did not cause regression of iris neovascularization. Complications of silicone oil included corneal decompensation, lens opacification, intraocular pressure elevation, and hypotony.

Conjunctival pigmentation following pars plana vitrectomy.

A 32-year-old white man developed distinctive episcleral and conjunctival pigmentation surrounding a sclerotomy site 6 weeks after vitrectomy and retinal reattachment surgery. The conjunctival lesion was biopsied during a second operation to remove an epiretinal membrane causing macular pucker. Light and electron microscopic studies demonstrated melanin granules extracellularly and within macrophages.

Acute retinal necrosis syndrome following herpes zoster dermatitis.

The acute retinal necrosis (ARN) syndrome has been recently linked to intraocular infection with one or more members of the herpesvirus family. The authors report two cases of ARN following herpes zoster skin eruptions, and one case following ipsilateral facial nerve palsy (Ramsay Hunt syndrome). Evaluation of serial serum antibody titers against cytomegalovirus (CMV), herpes simplex virus (HSV) (types 1 and 2), and varicella zoster (VZ) virus revealed diagnostic changes for VZ virus alone following the retinitis. Immune precipitation of radiolabeled VZ proteins by these sera followed by gel fractionation yielded radioimmune precipitation profiles characteristic of a recent zoster reactivation. These cases further implicate a central role for VZ virus infection in the etiology of the ARN syndrome.