Skeletal stem cell fate defects caused by Pdgfrb activating mutation.

Autosomal dominant PDGFRß gain-of-function mutations in mice and humans cause a spectrum of wasting and overgrowth disorders afflicting the skeleton and other connective tissues, but the cellular origin of these disorders remains unknown. We demonstrate that skeletal stem cells (SSCs) isolated from mice with a gain-of-function D849V point mutation in PDGFRß exhibit colony formation defects that parallel the wasting or overgrowth phenotypes of the mice. Single-cell RNA transcriptomics with SSC-derived polyclonal colonies demonstrates alterations in osteogenic and chondrogenic precursors caused by PDGFRßD849V. Mutant cells undergo poor osteogenesis in vitro with increased expression of Sox9 and other chondrogenic markers. Mice with PDGFRßD849V exhibit osteopenia. Increased STAT5 phosphorylation and overexpression of Igf1 and Socs2 in PDGFRßD849V cells suggests that overgrowth in mice involves PDGFRßD849V activating the STAT5-IGF1 axis locally in the skeleton. Our study establishes that PDGFRßD849V causes osteopenic skeletal phenotypes that are associated with intrinsic changes in SSCs, promoting chondrogenesis over osteogenesis.

Optimization of spray drying process of Japanese apricot (Prunus mume Sieb. et Zucc.) juice powder using nondigestible maltodextrin by response surface methodology (RSM).

The optimal spray-drying conditions for manufacturing Japanese apricot (Prunus mume Sieb. et Zucc.) juice powder (JAJP) using response surface methodology (RSM) were investigated. The optimization was performed using two independent factors, which are inlet air temperature (130-180 °C) and different concentrations of nondigestible maltodextrin (NMD) as a carrier agent (10-30%). Responses such as drying yield, moisture content, water solubility index (WSI), bulk density, color, pH, and antioxidant activities of JAJP were investigated. Moisture content, vitamin C content, color, antioxidant activity, pH and bulk density were greatly influenced by inlet air temperature, but dry yield and WSI were only significantly affected by NMD concentration. The optimum spray drying conditions were determined as 14.7% NMD concentration and 154.5 °C inlet air temperature, respectively. At these optimum conditions, a drying yield of 55.73%, 4.84% moisture content, 90.98% WSI, 0.59 g/mL of bulk density, and 169.87 mg/g vitamin C content in JAJP were measured. Therefore, JAJP with the desirable physicochemical properties could be produced.

Microbial communities of a variety of cheeses and comparison between core and rind region of cheeses.

Understanding the microbial community of cheese is important in the dairy industry, as the microbiota contributes to the safety, quality, and physicochemical and sensory properties of cheese. In this study, the microbial compositions of different cheeses (Cheddar, provolone, and Swiss cheese) and cheese locations (core, rind, and mixed) collected from the Arbuthnot Dairy Center at Oregon State University were analyzed using 16S rRNA gene amplicon sequencing with the Illumina MiSeq platform (Illumina, San Diego, CA). A total of 225 operational taxonomic units were identified from the 4,675,187 sequencing reads generated. Streptococcus was observed to be the most abundant organism in provolone (72 to 85%) and Swiss (60 to 67%), whereas Lactococcus spp. were found to dominate Cheddar cheese (27 to 76%). Species richness varied significantly by cheese. According to alpha diversity analysis, porter-soaked Cheddar cheese exhibited the highest microbial richness, whereas smoked provolone cheese showed the lowest. Rind regions of each cheese changed color through smoking and soaking for the beverage process. In addition, the microbial diversity of the rind region was higher than the core region because smoking and soaking processes directly contacted the rind region of each cheese. The microbial communities of the samples clustered by cheese, indicated that, within a given type of cheese, microbial compositions were very similar. Moreover, 34 operational taxonomic units were identified as biomarkers for different types of cheese through the linear discriminant analysis effect size method. Last, both carbohydrate and AA metabolites comprised more than 40% of the total functional annotated genes from 9 varieties of cheese samples. This study provides insight into the microbial composition of different types of cheese, as well as various locations within a cheese, which is applicable to its safety and sensory quality.

Temporal control of PDGFRα regulates the fibroblast-to-myofibroblast transition in wound healing.

Fibroblasts differentiate into myofibroblasts by acquiring new contractile function. This is important for tissue repair, but it also contributes to organ fibrosis. Platelet-derived growth factor (PDGF) promotes tissue repair and fibrosis, but the relationship between PDGF and myofibroblasts is unclear. Using mice with lineage tracing linked to PDGF receptor α (PDGFRα) gene mutations, we examine cell fates during skin wound healing. Elevated PDGFRα signaling increases proliferation but unexpectedly delays the fibroblast-to-myofibroblast transition, suggesting that PDGFRα must be downregulated for myofibroblast differentiation. In contrast, deletion of PDGFRα decreases proliferation and myofibroblast differentiation by reducing serum response factor (SRF) nuclear localization. Consequences of SRF deletion resemble PDGFRα deletion, but deletion of two SRF coactivators, MRTFA and MRTFB, specifically eliminates myofibroblasts. Our findings suggest a scenario where PDGFRα signaling initially supports proliferation of fibroblast progenitors to expand their number during early wound healing but, later, PDGFRα downregulation facilitates fibroblast differentiation into myofibroblasts.

Optimization of spray drying process parameters for production of Japanese apricot (Prunus mume Sieb. et Zucc.) juice powder.

Optimization of spray drying conditions namely inlet air temperature (IAT) and maltodextrin (MD) concentration was utilized by response surface methodology for Japanese apricot (Prunus mume Sieb. et Zucc.) juice powder (JAJP) manufacture. Drying yield, moisture content, water solubility index (WSI), bulk density, color, pH, total phenol content (TPC), total flavonoid content (TFC), vitamin C content, and DPPH radical-scavenging activity of juice powder were measured. Moisture content, vitamin C content, color, DPPH radical-scavenging activity, pH, and bulk density were greatly influenced by IAT, but drying yield, WSI, TPC, and TFC were only significantly affected by MD concentration. The spray drying condition was optimum at 10% MD concentration and 165.8 °C IAT. The properties of juice powder were 37.50% drying yield, 4.81% moisture content, 134.25 mg/g vitamin C content, 27.52% DPPH radical-scavenging activity, 2.78 pH, 89.15% WSI, 232.856 µg GAE/100 g TPC, 404.66 µg CE/100 g TFC, and 0.49 bulk density. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00950-8.

Incidence and Risk of Scleral-Fixated Akreos (AO60) Lens Opacification: A Case Series.

Purpose: Postoperative hydrophilic intraocular lens opacification can lead to decreased vision and may require intraocular lens exchange. This study aims to identify the incidence of scleral-fixated Akreos AO60's (Bausch + Lomb) lens opacification and risk factors for this phenomenon. Methods: This is a retrospective case series of all patients who underwent scleral-fixated Akreos AO60 lens at our institution between January 1, 2015 and December 31, 2019. The following data were recorded: age, sex, medical history, indication for Akreos AO60 implantation, laterality, ocular history, previous ocular surgical procedures, subsequent intraocular surgical procedures after the Akreos implantation, lens opacification, visual significance of opacification, and Akreos explantation. Intraoperative and postoperative complications were recorded. Main outcome measures were the overall incidence of Akreos lens opacification as well as the incidence of these eyes undergoing subsequent intraocular surgery. Results: A total of 262 eyes of 257 patients underwent Akreos lens implantation. Overall, 2% (5 of 262) developed lens opacification. Two patients had Descemet stripping automated endothelial keratoplasty (DSAEK) concurrently with Akreos implantation. One patient underwent subsequent Baerveldt glaucoma implantation and DSAEK. The fourth patient had vitrectomy with sulfur hexafluoride gas followed by DSAEK. This represents a 25% (4 of 16) opacification rate among all patients who underwent DSAEK (P ≤ .01, Fisher exact test). One patient developed opacification after undergoing 2 vitrectomies for retinal detachment in the absence of DSAEK. Conclusions: Akreos lens opacification can be visually significant and may occur after a retinal or corneal procedure that involves the use of intraocular gas or air.

Role of Holmium laser enucleation of the prostate to increase cancer detection rate in patients with gray-zone PSA level.

BACKGROUND: Even though the safety of the treatment for prostate cancer diagnosed by HoLEP has been reported, the diagnostic value of HoLEP for prostate cancer detection has not been confirmed. Therefore, we investigated the diagnostic potential of HoLEP for detecting prostate cancer. METHODS: Between December 2009 and October 2015, 359 patients (median age, 70.9 years; range, 66.2-74.8) were treated simultaneously with HoLEP and transrectal prostate needle biopsy (TPNB). Of these, 199 patients with a normal digital rectal examination and serum PSA concentration between 3.5 and 10.0 ng/mL were included in the study. Univariate and multivariate logistic regression analyses were performed to identify the predictive factor for prostate cancer detected by HoLEP. RESULTS: Median PSA, prostate volume and PSA density were 4.97 ng/mL (range, 4.20-6.70), 57.40 gm (range, 43.67-77.80) and 0.09 ng/mL2 (range, 0.07-0.12), respectively. Prostate cancer (Gleason score ≥6) was detected in 46 cases (23.1%). Of these, 26 (56.5%) were detected by HoLEP pathology, 11 (23.9%) by TPNB pathology, and 9 (19.6%) by both. Univariate and multivariate logistic regression analyses were performed in 179 patients, including benign prostatic hyperplasia patients (N=153, 76.9%) and patients with cancer detected by HoLEP pathology. PSA density was identified as an independent predictor of prostate cancer detected by HoLEP in gray-zone PSA. CONCLUSIONS: HoLEP is a viable modality for detecting prostate cancer in selected cases. PSA density was an independent predictor of prostate cancer detected by HoLEP in gray-zone PSA.

Inhibitory roles of peripheral nitrergic mechanisms in capsaicin-induced detrusor overactivity in the rat.

OBJECTIVES: To investigate the peripheral role of nitric oxide (NO) in capsaicin-induced detrusor overactivity (DO), as exogenously applied vanilloids can evoke NO release in urothelial cells but its functional role has not yet been reported. MATERIALS AND METHODS: The effects of N(G)-nitro-l-arginine methyl ester (L-NAME), an inhibitor of NO synthase (NOS), on bladder activity during intravesical capsaicin (30 microm) instillation were examined by using continuous infusion cystometry in urethane-anaesthetized rats. L-NAME was administered intravenously (i.v., 20 mg/kg), intrathecally (i.t., 270 microg/rat), intracerebroventricularly (i.c.v., 270 microg/rat) or intravesically (10 mg/mL) before or during capsaicin instillation. RESULTS: During cystometry with intravesical saline infusion, L-NAME injected i.v., i.t. and i.c.v., but not intravesically, significantly increased the intercontraction intervals (ICI) and L-NAME injected i.v., but not i.t., i.c.v. or intravesically, increased the maximum voiding pressure (MVP) without affecting the baseline pressure. Capsaicin instillation induced DO evidenced by a significant reduction in the ICI. L-NAME administered i.v. further decreased the ICI and increased the MVP and the baseline pressure during capsaicin instillation. Co-intravesical application of capsaicin and L-NAME also similarly enhanced capsaicin-induced DO. However, L-NAME injected i.t. or i.c.v. had no effect on capsaicin-induced DO. The excitatory effects of i.v and intravesical L-NAME on the ICI, MVP and baseline pressure during capsaicin infusion were significantly suppressed by desensitization of C-fibre afferent pathways by capsaicin pretreatment (125 mg/kg s.c., 4 days before cystometry). CONCLUSION: These results indicate that locally released NO can suppress DO induced by capsaicin-mediated C-fibre activation and that central NO pathways are not involved in capsaicin-induced DO.

Improvement in erectile dysfunction after neurotrophic factor gene therapy in diabetic rats.

PURPOSE: Erectile dysfunction (ED) is a common but difficult to treat complication of diabetes mellitus (DM). We have previously reported herpes simplex virus (HSV) vector mediated delivery of nerve growth factor into the bladder to treat diabetic cystopathy and neurotrophin-3 (NT3) gene transfer for pyridoxine treatment. Nerve growth factor and NT3 are neurotrophic factors that may protect nerves from mechanical and metabolic damage. We investigated the effects of HSV mediated delivery of NT3 for the treatment of diabetic ED. MATERIALS AND METHODS: Male Sprague-Dawley rats weighing 300 to 400 gm were injected with 65 mg/kg streptozotocin to induce DM. After 4 weeks 20 microl containing 5 x 10 pfu replication defective HSV vector expressing lacZ (6 rats) or NT3 (6) were injected directly into the cavernous nerve sheath with a 30 gauge needle. Four weeks later the animals underwent measurement of intracavernous pressure under electrical stimulation (20 Hz, 0.5 millisecond and 10 V) of the cavernous nerve. Staining for lacZ and neuronal nitric oxide synthase in the major pelvic ganglia was also performed. RESULTS: beta-Galactosidase staining revealed lacZ positive neurons in the major pelvic ganglia. Maximal intracavernous pressure induced by electrical stimulation showed statistically significant mean values +/- SEM of 15.1 +/- 2.1 and 43.8 +/- 11.1 cm H2O in the lacZ and NT3 vector injected groups, respectively (p = 0.03). The mean number of neuronal nitric oxide synthase positive neurons per section in the NT3 group was significantly higher than that in the lacZ control group at 3.33 +/- 0.23 and 0.64 +/- 0.14 neurons per high power field, respectively (p < 0.001). CONCLUSIONS: We report that gene therapy for the treatment of diabetic ED is feasible with HSV vectors. NT3 gene therapy may be applicable for the treatment of ED induced by DM.