bluuebunny/crossref_metadata_2025 huggingface dataset:
Cloning and sequence analysis of b-tubulin gene of Trypanosoma evansi isolates from Indian dromedaries (Camelus dromedarius)
Authors: Ashraf M. , Srivastava Meera , Ghorui S. K. , Nagarajan G. , Swami Shelesh Kumar , Kumar Sanjay
Published: September 2015

Trypanosomosis caused by Trypanosoma evansi is a disease of economic significance of domestic livestock, causing trypanosomiasis, commonly known as Surra. In the present study, gene of Trypanosoma evansi have beer cloned and sequenced. Beta-tubulin is important for cellular structure and physical functions. Sequence analysis of this gene will be useful for the vaccine studies. The open reading frame of b-tubulin of Trypanosoma evansi is 1329 bp in length, encoding 442 amino acid polypeptide. The sequence analysis revealed that the ß tubulin gene of Trypanosoma evansi from India was having the identity of 99% with T.equiperdum, T.evansi (China isolate), T.evansi (Indian isolates), T.brucei and 88% with T.cruzi. The stable biological characteristics of Trypanosomatidae beta-tubulin make it an ideal target of many anti-parasitic drugs that block tubulin function.
Effect of different levels of organic acids in the diets of hens on laying performance, egg quality criteria, blood parameters, and intestinal histomorphology
Authors: Kaya A. , Kaya H. , Gül M. , Yildirim B Apaydin , Timurkaan S.
Published: September 2015

The objective of the current study was to evaluate the effects of different levels (1.5, 3.0, 4.5 g/kg) of organic acid mixture (OAM) (60% formic acid, 20% propionic acid, and 20% soft acid) supplementation in the diets of laying hens on egg production, egg quality parameters, and intestinal histomorphology. Seventy-two Lohman strains of layers (26-week-old) were divided into four treatment groups. The hens were fed either a control diet or the control diet supplemented with 0, 1.5, 3 or 4.5 kg/t level of OAM from 26 to 38 weeks of age. The OAM supplementation did not affect feed consumption, egg production, egg weight, feed conversion ratio and body weight. Except for yolk index none of egg quality parameters and metabolic profile were affected by the dietary treatments. Inclusion of OAM into the basal diet increased villus height (quadraticaly), villus weight (quadraticaly), and tunica mucosal width (linearly) eexcept for crypt depth. Based on the results, dietary supplementation with OAM containing 60% formic acid, 20% propionic acid, and 20% soft acid appears to exert neither positive nor negative effects on laying performance, egg quality parameters, or serum blood parameters. But the OAM supplementation into basal diet had a positive effect on the intestinal histomorphology except for crypt depth.
Effect of organic selenium supplementation in the diets of finishing sheep on meat color and pH during shelf life
Authors: Libién-Jiménez Yamel , Mariezcurrena-Berasain María D. , Fuente Jorge Lugo De la , Kholif Abdelfattah Z. M. Salem Ahmed E. , Vaca-Paulín Rocío , Mariezcurrena-Berasain María A.
Published: September 2015

Supplementation of Selenium (Se) can improve the oxidative stability of meat products and retard metmyoglobin formation; so prolonging color. Effect of organic Se addition in the diets of sheep on meat stability; color and pH are limited and need to be studied. A study was conducted with eighteen Pelibuey female sheep at the finishing stage, and supplemented with organic Se-enriched Saccharomyces cerevisiae for sixty days to evaluate its effect on color and pH of Longissimus dorsi muscle. The research was conducted in a block randomized design considering three treatments; control (Se0) without the addition of yeast, or with 0.35 ppm of yeast (Se34) or with 0.60 ppm (Se59). Sheep were slaughtered at an average weight of 39.5 ± 4.41 kg. Meat color and pH were recorder in the cold carcass, 24 h after slaughtering, and during shelf life at 0, 4, 6 and 8 days after slaughtering under refrigeration at 4 °C. No significant differences were observed (P>0.05) for meat color and pH characteristics due to treatment. Decreased redness (a*) and Chroma (C*) values due to storage time were observed; however, the yellowness (b*) and angle Hue were increased. It could be concluded that supplementation of Se-enriched yeast in finishing sheep with 0.35 ppm and 0.60 ppm has no effect on meat color and pH characteristics.
Effect of dietary esterified glucomannan on performance of broilers exposed to aflatoxin
Authors: Arvind Maldhure Niteen , Churchil R. Richard
Published: September 2015

Effect of esterified glucomannan (E-GM), a new generation toxin binder enzymatically extracted from the cell wall of certain stains of Saccharomyces cerevisiae in counteracting the deleterious effects of aflatoxin in broiler chicken was investigated. Broilers fed 1ppm aflatoxin-B1 recorded significantly (Psignificantly (Ped the ill effects on weight gain and feed consumption, while the feed efficiency was restored to normal. High mortality in broilers caused by aflatoxin, was slightly reduced by the addition of E-GM.
Diagnostic reliability of enzymatic-immunoassayed serum pregnancy-associated glycoproteins in detecting early pregnancy compared to other screening measures in Aardi goats (Capra hircus) in saudi arabia
Authors: Al-Samawi Kamal A. , Al-Hassan Mohamed J. , Swelum Ayman
Published: September 2015

Despite proven ability of pregnancy-associated glycoproteins (PAGs) for early detection of pregnancy in several livestock, studies on its diagnostic reliability in Aardi goats are lacking. Thirty-two healthy mature Aardi does, managed under an intensive condition, were screened for pregnancy at 1, 7, 15, 23, 35 and 60 days post natural mating (NM) to evaluate the ability of utilizing caprine pregnancy-associated glycoproteins (caPAGs) compared to other methods frequently applied -but have several setbacks- in screening goats for early pregnancy diagnosis. According to the enzymatic-immunoassay data, our results indicated that pregnant Aardi does had higher (P 0.05) between the sensitivity, specificity, predictive values, and accuracy of serum caPAGs and other methods as early as day 23 post NM. Moreover, both serum caPAGs and US technique were proven to give higher (P y diagnosis in Aardi goats. In conclusion, the obtained results herein clearly demonstrated that enzymatic-immunoassayed serum caPAGs, as a direct, simple, accurate, inexpensive, practical and fast screening measure, was greatly feasible for distinguishing pregnant from non-pregnant as early as day 23 post NM, which is crucial to treat fertility problems early and efficiently.
Analgesic and adjunct actions of nalbuphine hydrochloride in xylazine or xylazine and acepromazine premedicated horses
Authors: Kulkarni Harish , William B. Justin , George Ravi Sundar , Kannan T. A.
Published: September 2015

The study was conducted in eighteen clinical cases of horses for diagnostic and surgical procedures requiring general anaesthesia were randomly divided into three groups, group I, group II and group III, each consisting of six cases. All the horses were premedicated with glycopyrrolate at the dose rate of 0.02 mg/kg body weight, intravenously. Horses in Group I and Group II were administered xylazine hydrochloride at the dose rate of 1.10 mg/kg body weight intravenously, whereas in Group III at the dose rate of 0.50 mg/kg body weight intravenously. In Group III, acepromazine was injected after xylazine administration, at the dose rate of 0.02mg/kg body weight, intravenously. Before induction of anaesthesia, nalbuphine hydrochloride was administered for Group II and Group III at the dose rate of 0.75 mg/kg body weight intravenously. Ketamine hydrochloride was administered intravenously to induce anaesthesia at the dose rate of 2.20 mg/kg body weight and maintained with 0.50 mg/kg body weight in required cases to maintain for duration of 15 ± 1.04 minutes. The mean time for induction in group I, group II and group III were 1.78 ± 0.27, 1.73 ± 0.10 and 1.85 ± 0.28 minutes respectively. The mean total number of additional doses of ketamine for standard duration of 15 ± 1.04 minutes surgery required in group I, group II and group III were 5.00 ± 0.36, 1.66 ± 0.33 and 2.00 ± 0.36 respectively. The quality of induction was 100 per cent smooth in group III, 83.33 per cent smooth and 16.67 per cent rough in group II and 66.66 per cent smooth and 33.34 per cent rough in group I. The quality of analgesia in group I, group II and group III were 2.83 ± 0.47, 1.83 ± 0.30 and 1.33 ± 0.21 respectively. The quality of muscle relaxation in group I, group II and group III were 3.16 ± 0.30, 1.50 ± 0.22 and 1.33 ± 0.21 respectively. The mean time for recovery in group I, group II and group III were 23.00 ± 1.52, 33.00 ± 0.93 and 41.98 ± 1.32 minutes respectively. The mean number of attempts for unassisted standing in group I, group II and group III were 6.66 ± 0.71, 5.00 ± 0.57 and 5.00 ± 0.36 respectively. The quality of recovery was 83.33 per cent smooth and 16.67 per cent rough in group III, 66.66 per cent smooth and 33.34 per cent rough in group II and 50.00 per cent smooth and 50.00 per cent rough in group I. None of the animals in any groups showed any intra and post operative complication.
Development of valuation index for draught cattle breeds of Tamil Nadu, India
Authors: Kumar N. K. Sudeep , Nisha P. R. , Kumar S. Senthil , Kumar G. Senthil
Published: September 2015

An attempt was made to develop a valuation index for draught cattle breeds of Tamil Nadu, which would help the farmers in getting the right price for their animals sold. The data were collected through personal interview using pre-tested interview schedule from 225 draught cattle owners from selected three districts of Tamil Nadu during the year 2010-11. A multiple linear regression was fitted for arriving at the valuation index of draught cattle breeds viz., Burgur, Kangayam and Umbalacherry. The adjusted R2 for overall draught cow valuation index was 0.818. The factors like presence of whirls in acceptable body region, skin and coat, body weight, milk yield and presence of pregnancy were found to be significant and had influenced the value of animal to the tune of Rs. 1642.40, Rs.1250.21, Rs. 36.25, Rs. 679.30 and Rs. 1241.24, respectively from their mean value. The overall draught bullock valuation index was also fitted and its adjusted R2 was 0.428. The variable body weight was found to be significant at one per cent level, whereas age was found to be significant at five per cent level and had negative influence on bullock value. The duration of work was found to be significant at five per cent level and it hiked the value at the rate of Rs.528.26. The variables like skin and coat, activeness, whirls, right pair, disease tolerance and breed were found to be non-significant.
Microbial quality of frozen squid (Sepioteuthis lessoniana, Lesson 1830) treated with food grade commercial chemicals
Authors: Kumar Vikas , Sukumar D. , Muruganantham M.
Published: September 2015

Squids perish rapidly like other fishes and require some treatment to maintain the quality for export. The microbial quality of frozen squid (Sepioteuthis lessoniana, Lesson 1830) treated with the commercial food grade chemicals imported from Spain was therefore studied. Dressed squids were treated with the chemicals Hidratech_4A (0.4%) and Whitech_3 (0.25%) dissolved in chilled freshwater (STCF) and chilled saltwater (STCS). Chilled squids not treated served as control (SNTC). Squids were quick frozen at -40°C in contact plate freezer and stored at-20±1°C. Samples were tested raw, after pre-processing, treatments and freezing and during storage at monthly intervals for 7 months. The microbial quality evaluation included Total Plate Count (TPC), Escherichia coli, Staphylococcus aureus, Vibrio cholerae and Salmonella. Study revealed a better quality of treated samples than control. Microbiological quality of STCS was better than STCF and SNTC. E. coli counts decreased after treatment. Salmonella and V. cholerae were absent.
Overcoming extinction: understanding processes of recovery of the Tibetan antelope
Authors: Leclerc C. , Bellard C. , Luque G. M. , Courchamp F.
Published: September 2015

Since the middle of the 20th century, the Tibetan antelope (Pantholops hodgsonii) has been poached for its wool to make luxury shawls, shahtoosh. This direct overexploitation caused a drastic decline in their population, with a loss of more than 90% compared to the baseline population a few decades ago. Assuming this is an anthropogenic Allee effect (AAE), human attraction for rarity can drive rare species to extinction, which could explain the increasing rates of antelope harvests, paralleling the escalating prices of shahtoosh as the species got rarer. Since 1999, international concern led to conservation actions and the population soon started increasing. This unique situation allowed the presence of an AAE in Tibetan antelope to be tested, as well as an assessment of the potential effects of conservation actions in the presence of this process. We developed a theoretical discrete‐time population dynamics model and examined effects of variation in shahtoosh prices. Furthermore, we tested the effects of major conservation actions into our models assessing their relative contribution to population recovery. During the exploitation phase, we found some evidence supporting the presence of an AAE compared to non‐AAE models when hunting ceased at antelope population sizes below 10% of the initial population size. Regarding the conservation phase, our results suggested that poacher numbers reduction had the most likely positive impact on Tibetan antelope population dynamics. As many other species are similarly declining and/or close to extinction, our results suggest that adequate conservation actions could counter the most dramatic declines, even when populations have entered an extinction vortex.
Caprock Analysis—A Practical Approach
Authors: Miller Guindon Leah
Published: September 2015

Caprock-integrity issues have been brought to the forefront with recent documented steam and bitumen leaks making their way to the ground surface, forcing the Alberta Energy Regulator (AER) to place a moratorium on the licensing of shallow thermal in-situ oil-sand projects. Reviewing core-acquisition, -storage, -analysis, and -preservation techniques can prevent potential project failure, capital loss, environmental liabilities, and public-safety risks. Responsible development of thermal projects requires that caprock integrity be quantified.
Stability of bicalicene isomers: A topological study
Authors: Gutman Ivan
Published: September 2015

Bicalicene is the conjugated hydrocarbon obtained by joining two calicene
fragments. This can be done in two different ways, thus resulting in two
bicalicene isomers (tentatively referred to as cis and trans). The trans
isomer is a stable compound whereas cis-bicalicene appears to be less stable
and has never been prepared. The stability order of the bicalicene isomers
cannot be rationalized by means of standard topological theory of conjugated
?-electron systems, and requires a special graph-theory-based analysis.
Factors associated with pathological fractures in dogs with appendicular primary bone neoplasia: 84 cases (2007–2013)
Authors: Rubin Jacob A. , Suran Jantra N. , Brown Dorothy Cimino , Agnello Kimberly A.
Published: September 2015

Objective—To determine the incidence of pathological fractures associated with appendicular primary bone tumors in dogs managed medically and to identify potential risk factors at the time of radiographic diagnosis that may be associated with eventual pathological fracture.

Design—Retrospective case series.

Animals—84 dogs with primary long bone neoplasia treated medically.

Procedures—Medical records for dogs with a diagnosis of primary long bone neoplasia based on results of radiography that was confirmed subsequently at necropsy were reviewed. Owners elected medical treatment at a pain clinic. Data regarding clinical signs, diagnostic testing, pathological findings, and outcome were evaluated.

Results—84 dogs met study inclusion criteria with 85 limbs affected. Osteosarcoma was the most common tumor and was identified in 78 of 85 (91.8%) limbs. The median time from diagnosis to euthanasia was 111 days (range, 28 to 447 days). Pathological fractures were identified in 33 of 85 limbs (38%), with the femur most commonly affected, (8/14 [57.1%]), followed by the tibia (9/17 [52.9%]), humerus (10/27 [37%]), radius (5/25 [20%]), and ulna (1/2 [50%]). Logistic regression analysis indicated that tumors arising from long bones other than the radius had odds of eventual fracture 5.05 as great as the odds for tumors of the radius, and lytic tumors had odds of eventual fracture 3.22 as great as the odds for tumors that appeared blastic or mixed lytic-blastic.

Conclusions and Clinical Relevance—Results suggested that radial primary bone tumors were less likely and lytic tumors were more likely to fracture. The overall incidence of pathological fractures secondary to appendicular primary bone neoplasia in this study with patients treated by means of intensive management for bone pain was higher than previously reported.
Prevalence of and risk factors associated with ovine progressive pneumonia in Wyoming sheep flocks
Authors: Gerstner Shelley , Adamovicz Jeffrey J. , Duncan John V. , Laegreid William W. , Marshall Katherine L. , Logan James R. , Schumaker Brant A.
Published: September 2015

Objective—To determine the prevalence of antibodies against small ruminant lentivirus (SRLV), the causative agent of ovine progressive pneumonia (OPP), and to identify risk factors associated with OPP in Wyoming sheep flocks.

Design—Cross-sectional study.

Animals—1,415 sheep from 54 flocks in Wyoming.

Procedures—Flocks were surveyed as part of the National Animal Health Monitoring System (NAHMS) 2011 sheep study. Serum samples obtained from sheep in Wyoming were analyzed for anti-SRLV antibodies by use of a competitive-inhibition ELISA. The prevalence of seropositive animals overall and within each flock was calculated. Respective associations between flock OPP status and various demographic and management variables were assessed.

Results—The estimated prevalence of sheep seropositive for anti-SRLV antibodies and OPP-infected flocks in Wyoming was 18.0% and 47.5%, respectively. Within OPP-infected flocks, the prevalence of seropositive sheep ranged from 3.9% to 96%. Flocks maintained on nonfenced range were more likely to be infected with OPP than were flocks maintained on fenced range (OR, 3.4; 95% confidence interval, 1.1 to 10.7). The estimated prevalence of OPP-infected flocks in Wyoming did not vary substantially from that at the regional or national level reported in the NAHMS 2001 sheep study. Compared with results of the NAHMS 2011 sheep study, Wyoming producers were more familiar with OPP than were other US sheep producers, but only 61% of Wyoming producers surveyed reported being very or somewhat familiar with the disease.

Conclusions and Clinical Relevance—Results indicated that OPP is prevalent in many Wyoming sheep flocks, which suggested that continued efforts are necessary to increase producer knowledge about the disease and investigate practices to minimize economic losses associated with OPP.
Earthquake Disaster and Anesthesiologists’ service in Nepal
Authors: Pradhan Bishwas
Published: September 2015

Journal of Society of Anesthesiologists of Nepal 2015; 2(2): 39-40
Oral midazolam versus oral triclofos for sedation of children for computed tomography scan - a randomized clinical trial
Authors: Gupta Anushu , Pandey Maitree , Choudhry Lalita , Jain Aruna , Pemde Harish
Published: September 2015

Background: Effective and safe pediatric procedural sedation is still a concern especially in areas outside operation theatres. The aim of the study was to compare the efficacy and safety of oral triclofos and oral midazolam in children undergoing computed tomography.Methods: A prospective randomized double blind study was conducted in 100 children aged one to five years. Group-I (n=50) received oral triclofos 100 mg/kg and Group-II (n=50) oral midazolam 0.75 mg/kg. Both groups were given oral atropine 0.03 mg/kg and supplemented with intravenous midazolam upto 0.1 mg/kg in case of inadequate effect. Onset and duration of sedation, success for completion of procedure and time to recovery were noted. Student’s t test and Z test of proportions were used for statistical analysis.Results Majority of children 36(72%) in Group-I achieved Ramsay Sedation Score >4 as compared to 25(50%) in Group-II. Computed tomography scan could be successfully completed at comparable rate (52% vs 56%). Success rate improved to 96% vs 80% after supplementing intravenous midazolam in Group I & II respectively (p< 0.05). Onset (37.91minutes ± 7.96 vs 26 ± 10), duration of sedation ( 117.91minutes ± 72.41 vs 66.2minutes ± 33) were significantly shorter and recovery (98.19minutes ± 72.58 vs 47.4minutes ± 31.42) in Group I & II respectively was faster in children who received oral midazolam (p< 0.05).Conclusion We conclude that both drugs were equally effective and safe for computed tomography scan in children. However better recovery profile of midazolam makes it more suitable for day care procedures.Journal of Society of Anesthesiologists of Nepal 2015; 2(2): 41-45
A Comprehensive Assessment Tool for Patient Registry Software Systems: The CIPROS Checklist
Authors: Mansmann U. , Lindoerfer D.
Published: September 2015

Background: Patient registries are an important instrument in medical research. Often their structure is complex and their implementation uses composite software systems to meet the wide spectrum of challenges.

Objectives: For the implementation of a registry, there is a wide range of commercial, open source, and self-developed systems available and a minimal standard for the critical appraisal of their architecture is needed.

Methods: We performed a systematic review of the literature to define a catalogue of relevant criteria to construct a minimal appraisal standard.

Results: The CIPROS list is developed based on 64 papers which were found by our systematic review. The list covers twelve sections and contains 72 items.

Conclusions: The CIPROS list supports developers to assess requirements on existing systems and strengthens the reporting of patient registry software system descriptions. It can be a first step to create standards for patient registry software system assessments.
Human Amnion-Derived Stem Cells Have Immunosuppressive Properties on NK Cells and Monocytes
Authors: Li Jiali , Koike-Soko Chika , Sugimoto Jun , Yoshida Toshiko , Okabe Motonori , Nikaido Toshio
Published: September 2015

Human amnion-derived cells are considered to be a promising alternative cell source for their potential clinical use in tissue engineering and regenerative medicine because of their proliferation and differentiation ability. The cells can easily be obtained from human amnion, offering a potential source without medical intervention. It has been proven that human amnion-derived cells express immunosuppressive factors CD59 and HLA-G, implying that they may have an immunosuppressive function. To assess the immunosuppressive activity, we investigated the effect of human amnion-derived cells on NK cell and monocyte function. Amnion-derived cells inhibited the cytotoxicity of NK cells to K562 cells. The inhibition depended on the NK/amnion-derived cell ratio. The inhibition of NK cytotoxicity was recovered by continuous culturing without amnion-derived cells. The inhibition of NK cytotoxicity was related to the downregulation of the expression of the activated NK receptors and the production of IFN-γ, as well as the upregulation of the expression of IL-10 and PGE2 in human amnion-derived cells. The addition of antibody to IL-10 or PGE2 inhibitor tended to increase NK cytotoxicity. IL-10 and PGE2 might be involved in the immunosuppressive activity of amniotic cells toward NK cells. Amniotic cells also suppressed the activity of cytokine production in monocytes analyzed with TNF-α and IL-6. These data suggested that amniotic cells have immunosuppressive activity.
Comparison of Magnetic Intensities for Mesenchymal Stem Cell Targeting Therapy on Ischemic Myocardial Repair: High Magnetic Intensity Improves Cell Retention but Has no Additional Functional Benefit
Authors: Shen Yunli , Liu Xuebo , Huang Zheyong , Pei Ning , Xu Jianfeng , Li Zheng , Wang Yunkai , Qian Juying , Ge Junbo
Published: September 2015

Magnetic targeting has the potential to enhance the therapeutic effects of stem cells through increasing retention of transplanted cells. To investigate the effects of magnetic targeting intensities on cell transplantation, we performed different magnetic intensities for mesenchymal stem cell (MSC)-targeting therapy in a rat model of ischemia/reperfusion. Rat MSCs labeled with superparamagnetic oxide nanoparticles (SPIOs) were injected into the left ventricular (LV) cavity of rats during a brief aorta and pulmonary artery occlusion. The 0.15 Tesla (T), 0.3 T, and 0.6 T magnets were placed 0~1 mm above the injured myocardium during and after the injection of 1 × 106 MSCs. Fluorescence imaging and quantitative PCR revealed that magnetic targeting enhanced cell retention in the heart at 24 h in a magnetic field strength-dependent manner. Compared with the 0 T group, three magnetic targeting groups enhanced varying cell engraftment at 3 weeks, at which time LV remodeling was maximally attenuated, and the therapeutic benefit (LV ejection fraction) was also highest in the 0.3 T groups. Interestingly, due to the low MSC engraftment resulting from microvascular embolisms, the 0.6 T group failed to translate into additional therapeutic outcomes, though it had the highest cell retention. Magnetic targeting enhances cell retention in a magnetic field strength-dependent manner. However, too high of a magnetic intensity may result in microembolization and consequently undermine the functional benefits of cell transplantation.
Ex Vivo Expanded Allogeneic Mesenchymal Stem Cells with Bone Marrow Transplantation Improved Osteogenesis in Infants with Severe Hypophosphatasia
Authors: Taketani Takeshi , Oyama Chigusa , Mihara Aya , Tanabe Yuka , Abe Mariko , Hirade Tomohiro , Yamamoto Satoshi , Bo Ryosuke , Kanai Rie , Tadenuma Taku , Michibata Yuko , Yamamoto Soichiro , Hattori Miho , Katsube Yoshihiro , Ohnishi Hiroe , Sasao Mari , Oda Yasuaki , Hattori Koji , Yuba Shunsuke , Ohgushi Hajime , Yamaguchi Seiji
Published: September 2015

Patients with severe hypophosphatasia (HPP) develop osteogenic impairment with extremely low alkaline phosphatase (ALP) activity, resulting in a fatal course during infancy. Mesenchymal stem cells (MSCs) differentiate into various mesenchymal lineages, including bone and cartilage. The efficacy of allogeneic hematopoietic stem cell transplantation for congenital skeletal and storage disorders is limited, and therefore we focused on MSCs for the treatment of HPP. To determine the effect of MSCs on osteogenesis, we performed multiple infusions of ex vivo expanded allogeneic MSCs for two patients with severe HPP who had undergone bone marrow transplantation (BMT) from asymptomatic relatives harboring the heterozygous mutation. There were improvements in not only bone mineralization but also muscle mass, respiratory function, and mental development, resulting in the patients being alive at the age of 3. After the infusion of MSCs, chimerism analysis of the mesenchymal cell fraction isolated from bone marrow in the patients demonstrated that donor-derived DNA sequences existed. Adverse events of BMT were tolerated, whereas those of MSC infusion did not occur. However, restoration of ALP activity was limited, and normal bony architecture could not be achieved. Our data suggest that multiple MSC infusions, following BMT, were effective and brought about clinical benefits for patients with lethal HPP. Allogeneic MSC-based therapy would be useful for patients with other congenital bone diseases and tissue disorders if the curative strategy to restore clinically normal features, including bony architecture, can be established.
Pancreatic Islet-Like Three-Dimensional Aggregates Derived from Human Embryonic Stem Cells Ameliorate Hyperglycemia in Streptozotocin-Induced Diabetic Mice
Authors: Shim Joong-Hyun , Kim Jonghyun , Han Jiyou , An Su Yeon , Jang Yu Jin , Son Jeongsang , Woo Dong-Hun , Kim Suel-Kee , Kim Jong-Hoon
Published: September 2015

We previously reported the in vitro differentiation of human embryonic stem cells (hESCs) into pancreatic endoderm. Here we demonstrate that islet-like three-dimensional (3D) aggregates can be derived from the pancreatic endoderm by optimizing our previous protocol. Sequential treatment with Wnt3a, activin A, and noggin induced a transient upregulation of T and MixL1, followed by increased expression of endodermal genes, including FOXA2, SOX17, and CXCR4. Subsequent treatment with retinoic acid highly upregulated PDX1 expression. We also show that inhibition of sonic hedgehog signaling by bFGF/activin βB and cotreatment with VEGF and FGF7 produced many 3D cellular clusters that express both SOX17 and PDX1. We found for the first time that proteoglycans and vimentin+ mesenchymal cells were mainly localized in hESC-derived PDX1+ clusters. Importantly, treatment with chlorate, an inhibitor of proteoglycan sulfation, together with inhibition of Notch signaling significantly increased the expression of Neurog3 and NeuroD1, promoting a transition from PDX1+ progenitor cells toward mature pancreatic endocrine cells. Purified dithizone+ 3D aggregates generated by our refined protocol produced pancreatic hormones and released insulin in response to both glucose and pharmacological drugs in vitro. Furthermore, the islet-like 3D aggregates decreased blood glucose levels and continued to exhibit pancreatic features after transplantation into diabetic mice. Generation of islet-like 3D cell aggregates from human pluripotent stem cells may overcome the shortage of cadaveric donor islets for future cases of clinical islet transplantation.
Stem Cell Therapy for Corneal Regeneration Medicine and Contemporary Nanomedicine for Corneal Disorders
Authors: Hsu Chih-Chien , Peng Chi-Hsien , Hung Kuo-Hsuan , Lee Yi-Yen , Lin Tai-Chi , Jang Shih-Fan , Liu Jorn-Hon , Chen Yan-Ting , Woung Lin-Chung , Wang Chien-Ying , Tsa Ching-Yao , Chiou Shih-Hwa , Chen Shih-Jen , Chang Yuh-Lih
Published: September 2015

The ocular surface is the outermost part of the visual system that faces many extrinsic or intrinsic threats, such as chemical burn, infectious pathogens, thermal injury, Stevens–Johnson syndrome, ocular pemphegoid, and other autoimmune diseases. The cornea plays an important role in conducting light into the eyes and protecting intraocular structures. Several ocular surface diseases will lead to the neovascularization or conjunctivalization of corneal epithelium, leaving opacified optical media. It is believed that some corneal limbal cells may present stem cell-like properties and are capable of regenerating corneal epithelium. Therefore, cultivation of limbal cells and reconstruction of the ocular surface with these limbal cell grafts have attracted tremendous interest in the past few years. Currently, stem cells are found to potentiate regenerative medicine by their capability of differentiation into multiple lineage cells. Among these, the most common cell sources for clinical use are embryonic, adult, and induced stem cells. Different stem cells have varied specific advantages and limitations for in vivo and in vitro expansion. Other than ocular surface diseases, culture and transplantation of corneal endothelial cells is another major issue for corneal decompensation and awaits further studies to find out comprehensive solutions dealing with nonregenerative corneal endothelium. Recently, studies of in vitro endothelium culture and ρ-associated kinase (ROCK) inhibitor have gained encouraging results. Some clinical trials have already been finished and achieved remarkable vision recovery. Finally, nanotechnology has shown great improvement in ocular drug delivery systems during the past two decades. Strategies to reconstruct the ocular surface could combine with nanoparticles to facilitate wound healing, drug delivery, and even neovascularization inhibition. In this review article, we summarized the major advances of corneal limbal stem cells, limbal stem cell deficiency, corneal endothelial cell culture/transplantation, and application of nanotechnology on ocular surface reconstruction. We also illustrated potential applications of current knowledge for the future treatment of ocular surface diseases.
A Rotating Bioreactor for Scalable Culture and Differentiation of Respiratory Epithelium
Authors: Raredon Micha Sam Brickman , Ghaedi Mahboobe , Calle Elizabeth A. , Niklason Laura E.
Published: September 2015

Respiratory epithelium is difficult to grow in vitro, as it requires a well-maintained polarizing air–liquid interface (ALI) to maintain differentiation. Traditional methods rely on permeable membrane culture inserts, which are difficult to work with and are ill-suited for the production of large numbers of cells, such as the quantities required for cell-based clinical therapies. Herein, we investigate an alternative form of culture in which the cells are placed on a porous substrate that is continuously rolled, such that the monolayer of cells is alternately submerged in media or apically exposed to air. Our prototype bioreactor is reliable for up to 21 days of continuous culture and is designed for scale-up for large-scale cell culture with continuous medium and gas exchange. Normal human bronchial epithelial (NHBE) cells were cultured on an absorbent substrate in the reactor for periods of 7, 14, and 21 days and were compared to static controls that were submerged in media. Quantification by immunohistochemistry and quantitative PCR of markers specific to differentiated respiratory epithelium indicated increased cilia, mucous production, and tight junction formation in the rolled cultures, compared to static. Together with scanning electron microscopy and paraffin histology, the data indicate that the intermittent ALI provided by the rolling bioreactor promotes a polarized epithelial phenotype over a period of 21 days.
Pollen structure and development in Nymphaeales: Insights into character evolution in an ancient angiosperm lineage
Authors: Taylor Mackenzie L. , Cooper Ranessa L. , Schneider Edward L. , Osborn Jeffrey M.
Published: September 2015

PREMISE OF THE STUDY:

A knowledge of pollen characters in early‐diverging angiosperm lineages is essential for understanding pollen evolution and the role of pollen in angiosperm diversification. In this paper, we report and synthesize data on mature pollen and pollen ontogeny from all genera of Nymphaeales within a comparative, phylogenetic context and consider pollen evolution in this early‐diverging angiosperm lineage. We describe mature pollen characters for Euryale, Barclaya, and Nymphaea ondinea, taxa for which little to no structural data exist.

METHODS:

We studied mature pollen for all nymphaealean genera using light, scanning electron, and transmission electron microscopy. We reviewed published reports of nymphaealean pollen to provide a comprehensive discussion of pollen characters in water lilies.

KEY RESULTS:

Nymphaeales exhibit diversity in key pollen characters, including dispersal unit size, ornamentation, aperture morphology, and tapetum type. All Nymphaeales pollen are tectate‐columellate, exhibiting one of two distinct patterns of infratectal ultrastructure—a thick infratectal space with robust columellae or a thin infratectal space with thin columellae. All genera have pollen with a lamellate endexine that becomes compressed in the proximal, but not distal wall. This endexine ultrastructure supports the operculate hypothesis for aperture origin. Nymphaeaceae pollen exhibit a membranous granular layer, which is a synapomorphy of the family.

CONCLUSIONS:

Variation in pollen characters indicates that significant potential for lability in pollen development was present in Nymphaeales at the time of its divergence from the rest of angiosperms. Structural and ontogenetic data are essential for interpreting pollen characters, such as infratectum and endexine ultrastructure in Nymphaeales.
Diversity of fig glands is associated with nursery mutualism in fig trees
Authors: Souza Camila D. , Pereira Rodrigo A. S. , Marinho Cristina R. , Kjellberg Finn , Teixeira Simone P.
Published: September 2015

PREMISE OF THE STUDY:

Fig trees (Moraceae) have remarkable enclosed inflorescences called figs or syconia. The flowers are pollinated by host‐specific fig wasps that enter the fig to lay their eggs. This nursery pollination system is one of the most studied of tropical mutualism interactions, but the source of the volatiles that attract fig wasps to their specific host figs has not been confirmed. The fragrance is the basis of host selection and, therefore, of reproductive isolation among sympatric Ficus species. This study locates and characterizes the glands likely to be responsible for pollinator attraction and also protection from herbivory in the figs of nine Ficus species representing all the major lineages within the genus.

METHODS:

Figs with receptive pistillate flowers were examined using light and scanning electron microscopy. Tests for histolocalization of substances were employed to detect glandular activity throughout the figs.

KEY RESULTS:

A great diversity of glands is found throughout the fig, and for the first time, the sites producing fragrances are identified. Scent glands are present on the ostiolar bracts and the outer layers of the fig receptacle. Laticifers and phenolic‐producing idioblasts, epidermis, and trichomes associated with fig protection occur on the ostiolar bracts, the fig receptacle, and floral tissues.

CONCLUSIONS:

The volatiles produced by glands on the ostiolar bracts are candidate sources for the long‐distance attraction of pollinator fig wasps. Scent glands on the outer layers of the receptacle may also play a role in chemical perception of the figs or may be related to their protection. The high cost to the plants if the figs are eaten and the temperature conditions required for nursery pollination are likely the factors that led to the selection of phenolic glands and laticifers during the group's evolution.
Synthesis and structures of ruthenium–NHC complexes and their catalysis in hydrogen transfer reaction
Authors: Chen Chao , Lu Chunxin , Zheng Qing , Ni Shengliang , Zhang Min , Chen Wanzhi
Published: September 2015

Ruthenium complexes [Ru(L1)2(CH3CN)2](PF6)2 (1), [RuL1(CH3CN)4](PF6)2 (2) and [RuL2(CH3CN)3](PF6)2 (3) (L1= 3-methyl-1-(pyrimidine-2-yl)imidazolylidene, L2 = 1,3-bis(pyridin-2-ylmethyl)benzimidazolylidene) were obtained through a transmetallation reaction of the corresponding nickel–NHC complexes with [Ru(p-cymene)2Cl2]2 in refluxing acetonitrile solution. The crystal structures of three complexes determined by X-ray analyses show that the central Ru(II) atoms are coordinated by pyrimidine- or pyridine-functionalized N-heterocyclic carbene and acetonitrile ligands displaying the typical octahedral geometry. The reaction of [RuL1(CH3CN)4](PF6)2 with triphenylphosphine and 1,10-phenanthroline resulted in the substitution of one and two coordinated acetonitrile ligands and afforded [RuL1(PPh3)(CH3CN)3](PF6)2 (4) and [RuL1(phen)(CH3CN)2](PF6)2 (5), respectively. The molecular structures of the complexes 4 and 5 were also studied by X-ray diffraction analysis. These ruthenium complexes have proven to be efficient catalysts for transfer hydrogenation of various ketones.
Comparison of approaches to deal with matrix effects in LC-MS/MS based determinations of mycotoxins in food and feed
Authors: Fabregat-Cabello N. , Zomer P. , Sancho J.V. , Roig-Navarro A.F. , Mol H.G.J.
Published: September 2015

This study deals with one of the major concerns in mycotoxin determinations: the matrix effect related to LC-MS/ MS systems with electrospray ionization sources. To this end, in a first approach, the matrix effect has been evaluated in two ways: monitoring the signal of a compound (added to the mobile phase) during the entire chromatographic run, and by classical post-extraction addition. The study was focused on nine selected mycotoxins: aflatoxin B1, fumonisins B1, B2 and B3, ochratoxin A, deoxynivalenol, T-2 and HT-2 toxins and zearalenone in various sample extracts giving moderate to strong matrix effects (maize, compound feed, straw, spices). Although the permanent monitoring of a compound provided a qualitative way of evaluating the matrix effects at each retention time, we concluded that it was not adequate as a quantitative approach to correct for the matrix effect. Matrix effects measured by post-extraction addition showed that the strongest ion suppression occurred for the spices (up to -89%). Five different calibration approaches to compensate for matrix effects were compared: multi-level external calibration using isotopically labelled internal standards, multi-level and single level standard addition, and two ways of single-point internal calibration: one point isotopic internal calibration and isotope pattern deconvolution. In general, recoveries and precision meeting the European Union requirements could be achieved with all approaches, with the exception of the single level standard addition at levels too close to the concentration in the sample. When an isotopically labelled internal standard is not available, single-level standard addition is the most efficient option.
Gene expression in the kidneys of broilers fed ochratoxin A for different time periods
Authors: Zeferino C.P. , Wells K.D. , Moura A.S.A.M.T. , Murarolli R.A. , Rottinghaus G.E. , Ledoux D.R.
Published: September 2015

Consumption of ochratoxin A (OTA) contaminated diets by broilers results in economic losses to the poultry industry. This experiment evaluated the effects of quantity and time of exposure to dietary OTA on performance, organ weights, serum biochemistry, and renal gene expression of chicks. Determination of genes expressed in response to OTA will allow for the identification of pathways that are influenced by OTA. 180-day old male broiler chicks were randomly assigned to a 3×3 factorial arrangement of treatments (3 levels of OTA; 0, 1 and 2 mg OTA/kg diet and 3 time periods; 7, 14 and 21 days) with 4 replicate pens of 5 birds each per treatment. For RNA-sequencing analysis (RNA-Seq), kidney samples were collected weekly from 3 controls and 3 chicks fed 1 mg OTA/kg. NextGENe software was used for read alignment and transcript quantification. Birds fed 2 mg OTA/kg diet had decreased feed intake and body weight gain, and increased serum uric acid on days 14 and 21. Compared to controls, birds fed 2 mg OTA/kg diet also had poorer feed conversion and increased kidney weights. On day 21, birds fed 1 mg OTA/kg diet had decreased albumin, and aspartate aminotransferase concentrations. Genes associated with carbohydrate and amino acid metabolism were downregulated, and genes associated with the immune system were upregulated at days 7 and 14. Genes associated with lipid metabolism and xenobiotic biodegradation were also downregulated on day 14. These changes disappeared on day 21 suggesting that the kidney and other related organs were repaired or the damage was contained. In conclusion, decreased performance and increased kidney weight and serum uric acid in birds fed 2 mg OTA/kg confirmed the effects of OTA. Supplementation of 1 mg OTA/kg diet caused time-dependent alterations in renal gene expression in chicks.
Efficacy of potential biocontrol agents for control ofFusarium verticillioides and fumonisin B1 under different environmental conditions
Authors: Samsudin N.I.P. , Magan N.
Published: September 2015

A mycotoxigenic strain ofFusarium verticillioides previously isolated from Malaysian maize kernels and identified morphologically and molecularly was used in the present work. The objectives were (1) to screen the competitiveness of three potential antagonists isolated from Malaysian maize and three other known candidates for control of growth ofF. verticillioides in vitro based on interaction scores, growth rates and hyphal area ofF. verticillioides, and (2) to examine the best candidates using different spore/cell ratios on milled maize agar at different water activity conditions on relative control of fumonisin B1 (FB1). Three fungi and three bacteria (BCAs 1-6) were examined for antagonistic effects againstF. verticillioides in dual-culture assays. These showed that all fungal candidates intermingled withF. verticillioides while all bacterial candidates inhibitedF. verticillioides on contact or at a distance, which in turn decreased the growth rates and hyphal area ofF. verticillioides significantly. Although BCA1 (Clonostachys rosea 016) did not inhibit growth or hyphal area ofF. verticillioides, it was included in FB1 inhibition studies with other bacterial candidates (BCA4,Streptomyces sp. AS1; BCA5, Gram-negative rod; BCA6,Enterobacter hormaechei) because of its established mycoparasitism. The FB1 inhibition studies were conducted on milled maize agar with different spore/cell ratios of pathogen:antagonist mixtures at 0.95/0.98 water activity (aw) and 25°C for 14 days. FB1 biosynthesis for all treatments was significantly higher at 0.95 than 0.98 aw. Of the four antagonists tested, the best was BCA1 which inhibited FB1 biosynthesis by 73 and 100% at 0.95 and 0.98 aw, respectively. BCA5 was the next best, resulting in 38 and 78% FB1 inhibition at 0.95 and 0.98 aw, respectively. The pathogen:antagonist ratios for BCA1 and BCA5 showed best results at 50:50 and 25:75. These results were discussed in context of using biocontrol agents to minimise fumonisins in maize.
Incarceration of umbilical hernia: a rare complication of large volume paracentesis
Authors: Khodarahmi Iman , Shahid Muhammad Usman , Contractor Sohail
Published: September 2015

We present two cases of umbilical hernia incarceration following large volume paracentesis (LVP) in patients with cirrhotic ascites. Both patients became symptomatic within 48 hours after the LVP. Although being rare, given the significantly higher mortality rate of cirrhotic patients undergoing emergent herniorrhaphy, this complication of LVP is potentially serious. Therefore, it is recommended that patients be examined closely for the presence of umbilical hernias before removal of ascitic fluid and an attempt should be made for external reduction of easily reducible hernias, if a hernia is present.
Implementación del levantamiento eficiente de fachadas mediante fotogrametría digital automatizada y el uso de software gratuito
Authors: Souto-Vidal M. , Ortiz-Sanz J. , Gil-Docampo M.
Published: September 2015

El levantamiento de fachadas es un proceso demandado en multitud de ámbitos como la planificación arquitectónica y el planeamiento. La fotogrametría ofrece gran precisión geométrica y nos permite visualizar la textura del objeto en cuestión. Con la aparición de software gratuito en este ámbito, ésta se convierte en una de las técnicas más asequibles. Para demostrar las posibilidades de esta metodología, se presentan aquí tres ejemplos de levantamientos de fachadas de distinta tipología, realizados con fotogrametría y mediante la utilización del software gratuito, utilizando para ello una cámara convencional sin calibrar.
Además se incluye un estudio en el que se muestra un análisis de la precisión alcanzable con este proceso, y una comparación directa con dos instrumentos topográficos convencionales. El trabajo realizado muestra, por tanto, como la fotogrametría de objeto cercano posibilita el levantamiento de fachadas a un bajo coste y de una forma ágil, precisa y eficiente.