Colletotrichum horii and Colletotrichum fioriniae Causing Anthracnose on Star Anise (Illicium verum) in China.

Star anise (Illicium verum) is an important economic and medical plant widely cultivated in Guangxi province, China. Its fruit can be used as spice and medicine (Wang et al. 2011). In recent years, anthracnose led to a serious decline in the production of star anise in Guangxi. In 2021, a survey conducted in CenwangLaoshan Reserve of Guangxi (24°21'N; 106°27'E) showed that the 2500 ha planting area had disease incidence greater than 80%. The leaf symptoms initially appeared as small spots, then expanded to round spots, finally becoming withered with grayish-white centers, surrounded by dark brown margins. Sometimes, small black acervuli were observed in the later stage. To explore the pathogen, infected leaves were collected and cut into small pieces (about 5 mm2) from the edge of the lesion, disinfected with 75% ethanol for 10 s, 1% NaClO for 1 min, washed with sterilized water and incubated on potato dextrose agar (PDA) plates at 28 °C in the dark. Ten single-spore isolates were obtained from the cultures. After 7 days on PDA at 28 °C, the colonies of 7 isolates were white with abundant aerial hyphae, gray-black with white-gray margins, and the other 3 isolates were light gray on the upper surface, and pink or orange on the underside. Representative isolates BS3-4 and BS3-1 were selected from 3 isolates and 7 isolates, respectively. Conidia of BS3-4 and BS3-1 were both hyaline, cylindrical, aseptate, smooth, apex obtuse, base truncate, and no significant differences (P > 0.05) in size between BS3-1 (13.22 to 5.38 × 3.89 to 1.99 µm) (n = 50) and BS3-4 (12.04 to 4.34 × 3.48 to 1.64 µm) (n = 50). These morphological characteristics were consistent with the Colletotrichum ssp. (Damm et al. 2012). The species identification of BS3-4 and BS3-1 was performed based on DNA sequence analysis. Genomic DNA was extracted as a template. Partial sequences of the rDNA internal transcribed spacer (ITS), actin gene (ACT), ß-tubulin2 (TUB2) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified and sequenced (Weir et al. 2012). The sequences were deposited in GenBank (ITS:OQ062642-43; ACT:OQ067614-15; GAPDH:OQ067616-17;TUB2：OQ067618-19). Based on the concatenated sequences of the 4 genes (ITS-ACT- GAPDH -TUB2) of BS3-4 and BS3-1 as well as sequences of other Colletotrichum spp. obtained from GenBank, the Maximum likelihood (ML) tree which produced with IQ-TREE (Minh et al. 2020) revealed that the isolate BS3-1 was Colletotrichum horii, and BS3-4 was Colletotrichum fioriniae. Pathogenicity was confirmed on healthy leaves of 1-year-old star anise seedlings (cultivar Dahong), and the leaves were wounded by sterilized toothpicks, and were inoculated with 10 µl of conidial suspensions of BS3-1 and BS3-4 (106 conidia/ml). Control seedlings were inoculated with sterilized distilled water. Five leaves per plant and 3 plants per treatment were selected. All inoculated seedlings were maintained in the greenhouse (12/12h light/dark, 25 ± 2℃, 90% relative humidity). Wound sites inoculated with BS3-1 and BS3-4 both turned greenish-brown after 2 days and then turned light brown with water-soaked spots. Black (BS3-1) or orange (BS3-4) dots of acervuli developed after 6 days. The lesion diameter of BS3-1 (14.4 mm) was larger than that of BS3-4 (8.1 mm). No symptoms were observed on controls. BS3-1 and BS3-4 were re-isolated from inoculated leaves, fulfilling Koch's postulates. Anthracnose of star anise caused by C.horii has been reported in China (Liao et al. 2017). However, to our knowledge, this is the first report of C.fioriniae infecting star anise in China. Accurate pathogen identification in this study could provide a reference for the control of anthracnose on star anise.

Overexpression and kinetic analysis of Ideonella sakaiensis PETase for polyethylene terephthalate (PET) degradation.

Ideonella sakaiensis PET hydrolase (IsPETase) is a well-characterized enzyme for effective PET biodegradation. However, the low soluble expression level of the enzyme hampers its practical implementation in the biodegradation of PET. Herein, the expression of IsPETaseMut, one of the most active mutants of IsPETase obtained so far, was systematically explored in E. coli by adopting a series of strategies. A notable improvement of soluble IsPETaseMut was observed by using chaperon co-expression and fusion expression systems. Under the optimized conditions, GroEL/ES co-expression system yielded 75 ± 3.4 mg·L-1 purified soluble IsPETaseMut (GroEL/ES), and NusA fusion expression system yielded 80 ± 3.7 mg·L-1 purified soluble NusA-IsPETaseMut, which are 12.5- and 4.6-fold, respectively, higher than its commonly expression in E. coli. The two purified enzymes were further characterized. The results showed that IsPETaseMut (GroEL/ES) displayed the same catalytic behavior as IsPETaseMut, while the fusion of NusA conferred new enzymatic properties to IsPETaseMut. Although NusA-IsPETaseMut displayed a lower initial hydrolysis capacity than IsPETaseMut, it showed a 1.4-fold higher adsorption constant toward PET. Moreover, the product inhibition effect of terephthalic acid (TPA) on IsPETase was reduced with NusA-IsPETaseMut. Taken together, the latter two catalytic properties of NusA-IsPETaseMut are more likely to contribute to the enhanced product release by NusA-IsPETaseMut PET degradation for two weeks.

Application of high expansion degradable cotton in nasal bleeding model of dog.

BACKGROUND: The discomfort and complications have always been problems for nasal packing materials. This study provided a new nasal packing material called high expansion degradable cotton (HEDC). METHODS: Nasal endoscopic surgery was used to establish a dog model of nasal bleeding, and wound surfaces were filled with Merocel, Nasopore and HEDC, respectively. Intraoperative and postoperative bleeding of 24 h was calculated. We evaluate the absorbability score, adhesion score, infection sore and nasal mucosal epithelium in postoperative 3, 7, 14 and 28 days. HE staining and electron microscopy were used to evaluate the recovery of nasal mucosa. RESULTS: There was no significant difference in nasal bleeding between HEDC, Merocel and Nasopore. Nasal endoscopic examination revealed HEDC absorbability of score, adhesion score, infection score were significantly lower than Merocel and Nasopore. The epithelialization time of HEDC was significantly shorter than that of Merocel and Nasopore. HE staining showed that HEDC and Nasopore could significantly reduce scar hyperplasia on the wound surface. The results of electron microscopy suggested that HEDC could protect the edge cilia of the wound. CONCLUSION: HEDC could be used as new choice for hemostasis after nasal endoscopic surgery, which could reduce nasal epithelialization time, and protect wound edge cilia.

Comparison of modification of a bacterial uricase with N-hydroxysuccinimide esters of succinate and carbonate of monomethoxyl poly(ethylene glycol).

Uricase after modification with monomethoxy poly(ethylene glycol) (mPEG) is currently the sole agent to treat refractory gout. For formulating Bacillus fastidious uricase, succinimidyl carbonate of mPEG-5000 (SC-mPEG5k) and succinimidyl succinate of mPEG-5000 (SS-mPEG5k) were compared. SC-mPEG5k possessed higher purity, comparable reaction rate constant with glycine but lower hydrolysis rate, and stronger effectiveness to modify amino groups. The uricase possessed two types of amino groups bearing a 25-fold difference in reactivity with SC-mPEG5k or SS-mPEG5k at pH 9.2. Oxonate and xanthine concentration-dependently protected the bacterial uricase from inactivation during PEGylation. With SC-mPEG5k at a molar ratio of 200 to uricase subunits and oxonate of 50 µM, the PEGylated uricase (1) retained about 73% of the original activity, (2) displayed about 10% reactivity to rabbit anti-sera recognizing the native uricase, (3) elicited IgG in rats accounting for about 5% of that by the native uricase, (4) exhibited circulation half-life time of about 25 H in cock plasma in vivo, and (5) concurrently maintained uric acid at lowered levels for over 20 H. Hence, PEGylation with SC-mPEG under the protection of a competitive inhibitor was a practical approach to formulation of the bacterial uricase; protection of enzymes by competitive inhibitors during PEGylation may have universal significance.

A first report of Streptococcus iniae infection of the spotted sea bass (Lateolabrax maculates).

This study marks the first occasion that Streptococcus iniae has been isolated, identified, and characterized as the causative pathogen in spotted sea bass (Lateolabrax maculates). Infected fish exhibited a range of external symptoms, including scale loss, bleeding from the jaw, anus, and tail, among other signs, as well as internal manifestations such as congested liver, splenomegaly, branchial anemia, yellow fat syndrome, and intestinal edema. Notably, exophthalmia and meningoencephalitis-typical symptoms associated with previous S. iniae infections-were not observed. A predominant bacterial isolate (designated 10S01) was recovered from the pure culture of spleen of a diseased spotted sea bass in Zhuhai, China. The strain was then subjected to Gram staining, biochemical profiling, and molecular confirmation through 16S rRNA and gyrB gene, corroborating its identity as S. iniae. Pathogenicity was assessed by intraperitoneal injection challenge in spotted sea bass weighing approximately 13 g/fish, revealing a LD50 of 74 cfu/g-fish. The 10S01 strain demonstrated the ability to colonize various organs, including the spleen, liver, kidney, and brain, with a relatively higher affinity for the spleen. Furthermore, antimicrobial susceptibility testing indicated that the 10S01 strain was sensitive to 14 tested antibiotics, particularly chloramphenicol, ciprofloxacin, clarithromycin, florfenicol, ofloxacin, rifampicin, and trimethoprim/sulfamethoxazole, highlighting these as preferred treatments for S. iniae infections in spotted sea bass. These findings contribute significantly to our understanding of S. iniae pathogenesis and inform the prompt and appropriate antibiotic treatment of S. iniae infections.

[Effects and Mechanisms of Polystyrene Microplastics on Extracellular Antibiotic Resistance Genes in Wastewater].

Microplastics (MPs) and antibiotic resistance genes (ARGs) are typical co-existing emerging pollutants in wastewater treatment plants. MPs have been shown to alter the distribution pattern of ARGs in sludge, but their effects on free extracellular ARGs (feARGs) in wastewater remain unclear. In this study, we used fluorescence quantitative PCR to investigate the dynamics of feARGs (including tetC, tetO, sul1, and sul2) in wastewater and their transition mechanisms after 60 d of exposure to typical MPs (polystyrene, PS). The results showed that the absolute abundance of tetracycline feARGs decreased by 28.4 %-76.0 % and 35.2 %-96.2 %, respectively, under nm-level and mm-level PS exposure and changed by -55.4 %-122.4 % under µm-level PS exposure. The abundance of sul1 showed a trend of nm-level > µm-level > mm-level upon PS exposure, and the changes in sul1 abundance was greater with ρ(PS)=50 mg·L-1 exposure. The relative abundance of sul2 was reduced by 25.4 %-42.6 % and 46.1 %-90.3 % after µm-level and mm-level PS exposure, respectively, and increased by 1.9-3.9 times after nm-level PS exposure, and the sul2 showed a higher reduction at ρ (PS)=50 mg·L-1 exposure than that at ρ (PS)=0.5 mg·L-1. The Pearson correlation analysis showed that the relative abundance of feARGs during PS exposure was positively correlated with cell membrane permeability and typical mobile genetic elements (intI1) abundance and negatively correlated with reactive oxygen species level. Our findings elucidated the effects and corresponding mechanisms of PS on the growth and mobility of feARGs in wastewater, providing a scientific basis for the control of the combined MPs and ARGs pollution in wastewater.

Comprehensive environmental performance of bottle-to-bottle recycling of PET bottles based on deposit-refund system in China.

Polyethylene terephthalate (PET) is a widely used packaging material and has high value in recycling. However, under China's dominant informal recycling system, most PET bottles are downcycled into fibers. The deposit-refund system (DRS) is considered a feasible mechanism to facilitate the high-value recycling of PET bottles. To comparatively evaluate the environmental performance [reduction of greenhouse gas (GHG) and pollutant emissions] under different scenarios using life cycle assessments, including the current system based on informal recycling, an improved system with a larger contribution from the source separation of municipal solid waste, and evolving systems with DRS application, five scenarios were set up. The DRS can reduce GHG emissions and the comprehensive environmental impact by 0.538 kg CO2 /kg PET bottles and 1.73 × 10-3 PE/kg PET bottles, respectively, compared to informal recycling. It can be concluded that the DRS-based recycling approach and the bottle-to-bottle recycling provide the substantial emission reduction potential of GHGs and pollutants.

Kinetics of Phosphate Ions and Phytase Activity Production for Lactic Acid-Producing Bacteria Utilizing Milling and Whitening Stages Rice Bran as Biopolymer Substrates.

A study evaluated nine kinetic data and four kinetic parameters related to growth, production of various phytase activities (PEact), and released phosphate ion concentration ([Pi]) from five lactic acid bacteria (LAB) strains cultivated in three types of media: phytate (IP6), milling stage rice bran (MsRB), and whitening stage rice bran (WsRB). Score ranking techniques were used, combining these kinetic data and parameters to select the most suitable LAB strain for each medium across three cultivation time periods (24, 48, and 72 h). In the IP6 medium, Lacticaseibacillus casei TISTR 1500 exhibited statistically significant highest (p ≤ 0.05) normalized summation scores using a 2:1 weighting between kinetic and parameter data sets. This strain also had the statistically highest levels (p ≤ 0.05) of produced phosphate ion concentration ([Pi]) (0.55 g/L) at 72 h and produced extracellular specific phytase activity (ExSp-PEact) (0.278 U/mgprotein) at 48 h. For the MsRB and WsRB media, Lactiplantibacillus plantarum TISTR 877 performed exceptionally well after 72 h of cultivation. It produced ([Pi], ExSp-PEact) pairs of (0.53 g/L, 0.0790 U/mgprotein) in MsRB and (0.85 g/L, 0.0593 U/mgprotein) in WsRB, respectively. Overall, these findings indicate the most promising LAB strains for each medium and cultivation time based on their ability to produce phosphate ions and extracellular specific phytase activity. The selection process utilized a combination of kinetic data and parameter analysis.

Trojan-Horse Diameter-Reducible Nanotheranostics for Macroscopic/Microscopic Imaging-Monitored Chemo-Antiangiogenic Therapy.

Although nanotheranostics have displayed striking potential toward precise nanomedicine, their targeting delivery and tumor penetration capacities are still impeded by several biological barriers. Besides, the current antitumor strategies mainly focus on killing tumor cells rather than antiangiogenesis. Enlightened by the fact that the smart transformable self-targeting nanotheranostics can enhance their targeting efficiency, tumor penetration, and cellular uptake, we herein report carrier-free Trojan-horse diameter-reducible metal-organic nanotheranostics by the coordination-driven supramolecular sequential co-assembly of the chemo-drug pemetrexed (PEM), transition-metal ions (FeIII), and antiangiogenesis pseudolaric acid B. Such nanotheranostics with both a high dual-drug payload efficiency and outstanding physiological stability are responsively decomposed into numerous ultra-small-diameter nanotheranostics under stimuli of the moderate acidic tumor microenvironment and then internalized into tumor cells through tumor-receptor-mediated self-targeting, synergistically enhancing tumor penetration and cellular uptake. Besides, such nanotheranostics enable visualization of self-targeting capacity under the macroscopic monitor of computed tomography/magnetic resonance imaging, thereby realizing efficient oncotherapy. Moreover, tumor microvessels are precisely monitored by optical coherence tomography angiography/laser speckle imaging during chemo-antiangiogenic therapy in vivo, visually verifying that such nanotheranostics possess an excellent antiangiogenic effect. Our work will provide a promising strategy for further tumor diagnosis and targeted therapy.

Performance of PROPELLER FSE T2WI in reducing metal artifacts of material porcelain fused to metal crown: a clinical preliminary study.

This study aimed to compare MRI quality between conventional fast spin echo T2 weighted imaging (FSE T2WI) with periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) FSE T2WI for patients with various porcelain fused to metal (PFM) crown and analyze the value of PROPELLER technique in reducing metal artifacts. Conventional FSE T2WI and PROPELLER FSE T2WI sequences for axial imaging of head were applied in participants with different PFM crowns: cobalt-chromium (Co-Cr) alloy, pure titanium (Ti), gold-palladium (Au-Pd) alloy. Two radiologists evaluated overall image quality of section in PFM using a 5-point scale qualitatively and measured the maximum artifact area and artifact signal-to-noise ratio (SNR) quantitatively. Fifty-nine participants were evaluated. The metal crown with the least artifacts and the optimum image quality shown in conventional FSE T2WI and PROPELLER FSE T2WI were in Au-Pd alloy, Ti, and Co-Cr alloy order. PROPELLER FSE T2WI was superior to conventional FSE T2WI in improving image quality and reducing artifact area for Co-Cr alloy (17.0 ± 0.2% smaller artifact area, p < 0.001) and Ti (11.6 ± 0.7% smaller artifact area, p = 0.005), but had similar performance compared to FSE T2WI for Au-Pd alloy. The SNRs of the tongue and masseter muscle were significantly higher on PROPELLER FSE T2WI compared with conventional FSE T2WI (tongue: 29.76 ± 8.45 vs. 21.54 ± 9.31, p = 0.007; masseter muscle: 19.11 ± 8.24 vs. 15.26 ± 6.08, p = 0.016). Therefore, the different PFM crown generate varying degrees of metal artifacts in MRI, and the PROPELLER can effectively reduce metal artifacts especially in the PFM crown of Co-Cr alloy.

The therapeutic effect of stem cells from human exfoliated deciduous teeth on a rat model of tracheal fistula.

BACKGROUND: Tracheal fistulas (TF) can be dangerous and even fatal in patients. The current treatment is really challenging. Previous studies reported that mesenchymal stem cells (MSCs) could be used to treat respiratory tract fistulas. Stem cells from human exfoliated deciduous teeth (SHED) are considered to be MSC-like cells that may also have the potential to treat the tracheal fistulas. In this study, we investigated the therapeutic effects of SHED in rat tracheal fistula models. METHODS: A total of 80 SD rats were randomly divided into five groups: a sham-operated group, a local PBS group (L-PBS), an intravenous PBS group (I-PBS), a local SHED treatment group (L-SHED), and an intravenous SHED treatment group (I-SHED). The L-SHED and I-SHED groups were given a topical application around the fistula or an intravenous injection of 1*107 SHED via the tail vein, respectively, while the L-PBS and I-PBS groups were given an equivalent volume of PBS through local or intravenous administration. A stereomicroscope was used to observe fistula healing on the 2nd, 3rd, and 5th days following transplantation. On the 7th day, the survival of SHED was observed by immunofluorescence. The pathology of the lungs and fistulas was observed by hematoxylin and eosin (H&E) and Masson staining. The expression levels of the Toll-like receptor 4 (TLR4), interleukin (IL)-1ß, IL-33, and IL-4 were measured using immunohistochemistry. The expression levels of TLR4, high mobility group box 1 (HMGB1), and myeloid differentiation factor 88 (MYD88) were studied using western blotting. On day 14, airway responsiveness of rats was detected and analyzed. RESULTS: Fistula healing in the L-SHED and I-SHED groups was faster than that in their respective PBS groups after transplantation. The fistula diameters in the L-SHED and I-SHED groups were significantly smaller than those in the L-PBS and I-PBS groups on the 3rd day. Moreover, the phenomenon of fibroblast proliferation and new blood vessel growth around the fistula seemed more pronounced in the L-SHED and I-SHED groups. Although no discernible difference was found in airway responsiveness after SHED treatment, the degree of inflammation in the lungs was reduced by intravenous SHED treatment. However, there was no significant reduction in lung inflammation by local SHED treatment. The expression levels of IL-1ß and IL-33 were decreased in the I-SHED group, while IL-4 was elevated compared with the I-PBS group. Interestingly, intravenous SHED treatment inhibited the activation of HMGB1/TLR4/MYD88 in the lung tissues of TF rats. CONCLUSIONS: SHED transplantation accelerated the rate of fistula healing in rats. Intravenous SHED treatment reduced lung inflammation. Thus, SHED may have potential in the treatment of tracheal fistula, providing hope for future therapeutic development for TF.

Missense mutation in DYNC1H1 gene caused psychomotor developmental delay and muscle weakness: A case report.

BACKGROUND: The DYNC1H1 gene encodes a part of the dynamic protein, and the protein mutations may further affect the growth and development of neurons, resulting in degeneration of anterior horn cells of the spinal cord, and a variety of clinical phenotypes finally resulting in axonal Charcot-Marie-Tooth disease type 20 (CMT20), mental retardation 13 (MRD13) and spinal muscular atrophy with lower extremity predominant 1 (SMA-LED). The incidence of the disease is low, and it is difficult to diagnose, especially in children. Here, we report a case of DYNC1H1 gene mutation and review the related literature to improve the pediatrician's understanding of DYNC1H1 gene-related disease to make an early correct diagnosis and provide better services for children. CASE SUMMARY: A 4-mo-old Chinese female child with adducted thumbs, high arch feet, and epileptic seizure presented slow response, delayed development, and low limb muscle strength. Electroencephalogram showed abnormal waves, a large number of multifocal sharp waves, sharp slow waves, and multiple spasms with a series of attacks. High-throughput sequencing and Sanger sequencing identified a heterozygous mutation, c.5885G>A (p.R1962H), in the DYNC1H1 gene (NM_001376) of the proband, which was not identified in her parents. Combined with the clinical manifestations and pedigree of this family, this mutation is likely pathogenic based on the American Academy of Medical Genetics and Genomics guidelines. The child was followed when she was 1 year and 2 mo old. The magnetic resonance imaging result was consistent with the findings of white matter myelinated dysplasia and congenital giant gyrus. The extensive neurogenic damage to the extremities was considered, as the results of electromyography showed that the motor conduction velocity and sensory conduction of the nerves of the extremities were not abnormal, and the degree of fit of the children with severe contraction was poor. At present, the child is 80 cm in length and 9 kg in weight, with slender limbs and low muscle strength, and still does not raise her head. She cannot sit or speak. Speech, motor, and mental development was significantly delayed. There is still no effective treatment for this disease. CONCLUSION: We herein report a de novo variant of DYNC1H1 gene, c.5885G>A (p.R1962H), leading to overlapping phenotypes (seizure, general growth retardation, and muscle weakness) of CMT20, MRD13, and SMA-LED, but there is no effective treatment for such condition. Our case enriches the DYNC1H1 gene mutation spectrum and provides an important basis for clinical diagnosis and treatment and genetic counseling.

Effects of modification of amino groups with poly(ethylene glycol) on a recombinant uricase from Bacillus fastidiosus.

After modification with monomethoxyl-poly(ethylene glycol)-5000, a recombinant intracellular uricase from Bacillus fastidiosus ATCC 29604 showed residual activity of about 65%, a thermo-inactivation half-life >85 h, a circulating half-life about 20 h in rats in vivo, consistent effects of common cations, and consistent optima for reaction temperature and pH. Thus, this uricase can be formulated via modification with monomethoxyl-poly(ethylene glycol).

Polystyrene microplastics increase uptake, elimination and cytotoxicity of decabromodiphenyl ether (BDE-209) in the marine scallop Chlamys farreri.

Microplastics are a growing problem in marine environments due to their ubiquitous occurrence and affinity for chemical pollutants. However, the influence of microplastics on the uptake, depuration and toxicity of decabromodiphenyl ether (BDE-209) in marine organisms is unclear. We exposed the marine scallop Chlamys farreri to polystyrene microplastics (PS; 125 µg/L) combined with BDE-209 (10 and 100 µg/L) to determine their toxicokinetics, cellular toxicity and histopathological effects. The results showed that PS acted as both a carrier and a scavenger for the bioaccumulation of BDE-209. Importantly, the carrier role of PS was greater than scavenger one. PS increased the negative effect of BDE-209 (100 µg/L) on hemocyte phagocytosis, and ultrastructural changes in gills and digestive gland of scallops due to their carrier role for the bioaccumulation of BDE-209. However, PS did not increase the DNA damage of BDE-209 on the hemocytes. These findings are evidence of microplastics transferring adsorbed pollutants to marine organisms, and increasing their toxicity.

Silicon-substituted hydroxyapatite composite coating by using vacuum-plasma spraying and its interaction with human serum albumin.

The incorporation of silicon can improve the bioactivity of hydroxyapatite (HA). Silicon-substituted HA (Ca(10)(PO(4))(6-x )(SiO(4))( x )(OH)(2-x ), Si-HA) composite coatings on a bioactive titanium substrate were prepared by using a vacuum-plasma spraying method. The surface structure was characterized by using XRD, SEM, XRF, EDS and FTIR. The bond strength of the coating was investigated and XRD patterns showed that Ti/Si-HA coatings were similar to patterns seen for HA. The only different XRD pattern was a slight trend toward a smaller angle direction with an increase in the molar ratio of silicon. FTIR spectra showed that the most notable effect of silicon substitution was that -OH group decreased as the silicon content increased. XRD and EDS elemental analysis indicated that the content of silicon in the coating was consistent with the silicon-substituted hydroxyapatite used in spraying. A bioactive TiO(2) coating was formed on an etched surface of Ti, and the etching might improve the bond strength of the coatings. The interaction of the Ti/Si-HA coating with human serum albumin (HSA) was much greater than that of the Ti/HA coating. This might suggest that the incorporation of silicon in HA can lead to significant improvements in the bioactive performance of HA.

Effect of Provisional Restorations with Ovate Pontics on Preservation of the Ridge after Tooth Extraction: Case Series.

OBJECTIVE: To evaluate the effect of provisional restorations using ovate pontics on the hard and soft tissues of the alveolar ridge after tooth extraction. METHODS: Patients with intact buccal bone after extraction of a maxillary incisor underwent a series of treatments. The protocol included tooth extraction and provisional restoration using ovate pontics. The outcomes were assessed 1, 3 and 6 months after the procedure, regarding changes in bone volume and soft tissue dimension. RESULTS: A total of 11 patients (6 female and 5 male) were treated using an ovate pontic provisional restoration after tooth extraction. After 6 months, bone loss at level 1, 3 and 5 mm below the most coronal section of the bone crest was 2.28 ± 0.53 mm, 1.51 ± 0.30 mm and 1.46 ± 0.41 mm, respectively. Changes in bone volume of these three levels were significantly lower (P < 0.01) than changes previously reported in the literature. The mesial and distal papillae recessed 0.47 ± 0.24 mm and 0.88 ± 0.57 mm, respectively, whereas the midfacial shrinkage was 0.86 ± 0.44 mm. The dimension of the ridge contour around the socket reduced mostly at 3 and 5 mm below the coronal section and the gingival mucosa, with values of 1.68 ± 0.48 mm and 1.61 ± 0.48 mm, respectively. CONCLUSION: Using a provisional restoration with ovate pontics after tooth extraction may preserve the alveolar ridge by sustaining the soft tissue, in particular the gingival papilla.

Expanding the clinical and genetic spectrum of Heimler syndrome.

BACKGROUND: Heimler syndrome (HS) is a rare hereditary systemic disorder, partial clinically overlapping with Usher syndrome. So far, our knowledge of HS is very limited, many cases are misdiagnosed or may not even be diagnosed at all. This study aimed to analyze the clinical and genetic characteristics of HS, and to evaluate potential phenotype-genotype correlations. RESULTS: Two HS cases caused by PEX1 mutations were identified, and a novel likely pathogenic mutation, PEX1 c.895_896insTATA, was found. The main ophthalmic finding of the two patients was consistent with retinitis pigmentosa accompanied by cystoid macular edema, but short axial length and hyperopia were also observed as two previously unreported ocular phenotypes. Analysis of the literature showed that of the 29 HS patients previously reported, 12 had PEX6 mutations, 10 had PEX1 mutations, two had PEX26 mutations, and the remaining patients were not genetically tested. Three novel genotype-phenotype correlations were revealed from analysis of these patients. First, most genotypes of every HS patient include at least one missense variant; second, at least one mutation in PEX1 or PEX6 gene affects the AAA-ATPase region in every HS patient with retinal dystrophy, suggesting AAA-ATPase region is a hypermutable region in patients with a retinal dystrophy; third, there are no significant differences between PEX1-, PEX6-, and PEX26-associated phenotypes. CONCLUSION: Next-generation sequencing is important for the diagnosis of HS. This study expands the clinical and genetic spectrum of HS, and provides additional insights into genotype-phenotype correlations, which is vital for accurate clinical practice, genetic counseling, and pathogenesis studies.

Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor.

A novel microfluidic device that can selectively and specifically isolate exceedingly small numbers of circulating tumor cells (CTCs) through a monoclonal antibody (mAB) mediated process by sampling large input volumes (>/=1 mL) of whole blood directly in short time periods (<37 min) was demonstrated. The CTCs were concentrated into small volumes (190 nL), and the number of cells captured was read without labeling using an integrated conductivity sensor following release from the capture surface. The microfluidic device contained a series (51) of high-aspect ratio microchannels (35 mum width x 150 mum depth) that were replicated in poly(methyl methacrylate), PMMA, from a metal mold master. The microchannel walls were covalently decorated with mABs directed against breast cancer cells overexpressing the epithelial cell adhesion molecule (EpCAM). This microfluidic device could accept inputs of whole blood, and its CTC capture efficiency was made highly quantitative (>97%) by designing capture channels with the appropriate widths and heights. The isolated CTCs were readily released from the mAB capturing surface using trypsin. The released CTCs were then enumerated on-device using a novel, label-free solution conductivity route capable of detecting single tumor cells traveling through the detection electrodes. The conductivity readout provided near 100% detection efficiency and exquisite specificity for CTCs due to scaling factors and the nonoptimal electrical properties of potential interferences (erythrocytes or leukocytes). The simplicity in manufacturing the device and its ease of operation make it attractive for clinical applications requiring one-time use operation.

The Effect of Chronic Fluorosis on Calcium Ions and CaMKIIα, and c-fos Expression in the Rat Hippocampus.

This study investigated neurotoxicity of chronic fluorosis in the rat hippocampus. Newly weaning, male, Sprague-Dawley (SD) rats were administered 15, 30, and 60 mg/L sodium fluoride (NaF) solution (fluorine ion concentration 8.25, 16.50, and 33.00 mg/L, respectively), and tap water, for 18 months. The neurotoxicological mechanism was examined with a focus on intracellular calcium overload. Results showed that as the fluoride concentration increased, calcium ion concentration [Ca2+], the expression of calcium/calmodulin-dependent protein kinase II α (CaMKIIα), and the expression of catus proto-oncogene protein c-fos (c-fos) all tend to increase. Compared to the control group, Ca2+, CaMKIIα, and c-fos significantly increased (P < 0.05) in the moderate-fluoride and the high-fluoride groups. These results indicate that Ca2+/CaMKIIα/c-fos channel signal may be the molecular mechanism of central nervous system damage caused by chronic fluoride intoxication. Moreover, elevated Ca2+ concentration in the hippocampus may be the initiating factor of neuronal apoptosis induced by fluoride.