bluuebunny/crossref_metadata_2025 huggingface dataset:
Reactive amyloidosis associated with ischial callosititis
Authors: Liu David X. , Gilbert Margaret H. , Wang Xiaolei , Didier Peter J. , Veazey Ronald S.
Published: October 2012

Ischial callosities have received little attention in veterinary medicine even though they are distinguishing anatomic organs. The organs are characterized by a pair of hairless pads of thickened epidermis, located bilaterally in the gluteal region, which overlay the tuberosities of the ischia of all Old World monkeys, gibbons, and siamangs. The current report describes a case of reactive amyloidosis associated with ischial callosititis in a rhesus macaque ( Macaca mulatta). Amyloid A (AA) protein was found in the liver, spleen, small intestine, mesenteric lymph nodes, and ischial callosities by histology, Congo red stain, and immunohistochemistry. Confocal microscopy showed that many cluster of differentiation (CD)68-positive macrophages within the ischial callosities contained intracellular AA protein, which suggests that CD68-positive macrophages have an important role in the pathogenesis of reactive amyloidosis in nonhuman primates. The normal histology of ischial callosities of rhesus macaques is also documented in this report.
Emerging tumefactive multiple sclerosis after switching therapy from natalizumab to fingolimod
Authors: Jander Sebastian , Turowski Bernd , Kieseier Bernd C , Hartung Hans-Peter
Published: October 2012

In this report we describe a multiple sclerosis patient who developed a relapse with magnetic resonance imaging (MRI) features of tumefactive demyelination after switching therapy from natalizumab to fingolimod. Tumefactive lesions emerged 16 weeks after stopping natalizumab and eight weeks after commencing fingolimod therapy but had been absent at the time of diagnosis and throughout the preceding course of the disease. Thus, the first-time occurrence of atypical lesion features may have been caused by the change in immunotherapy. The possible relevance of natalizumab withdrawal vs fingolimod introduction is discussed against the background of recently published case studies.
Switching therapy from natalizumab to fingolimod in relapsing–remitting multiple sclerosis: clinical and magnetic resonance imaging findings
Authors: Rinaldi F , Seppi D , Calabrese M , Perini P , Gallo P
Published: October 2012

Clinical and/or neuroimaging evidence of disease reactivation has been described in multiple sclerosis (MS) patients after a break from natalizumab. Whether fingolimod might be a therapeutic option following natalizumab needs to be evaluated. Twenty-two relapsing remitting MS patients having JC virus antibodies (JCVAb+) in serum were shifted from natalizumab to fingolimod after a three-month washout period. Neurological evaluation with the Expanded Disability Status Scale (EDSS) was performed monthly for a mean follow-up period of nine months. In 20/22 patients, brain magnetic resonance imaging (MRI) was obtained within one month after therapy initiation. Disease reactivation was observed in 11/22 (50%) patients: clinical relapses in six patients (four patients within the first month of therapy) and MRI activity in a further five patients (three patients within the first month of therapy). Clinical and/or MRI signs suggestive of disease rebound were observed in three patients. Our data indicate that fingolimod does not exert clinical activity quickly enough to stop MS reactivation after a break from natalizumab.
Using Research to Understand Youth in High-Risk Urban Communities
Authors: Yarmuth Megan , Patterson Jennifer , Burton Tessa , Douglas Caitlin , Taylor Trish , Boyle Marie
Published: October 2012

Research from the Centers for Disease Control and Prevention (CDC) shows that nearly 1.5 million high school students a year are affected by dating violence and that youth who are physically hurt by a boyfriend/girlfriend are more likely to report binge drinking, suicide attempts, and other harmful behaviors. Dating violence may be more prevalent in economically and socially disadvantaged communities, especially in urban areas. Targeting youth with prevention messages before they start dating may avert teen dating violence and subsequent adult intimate partner violence; however, there is a dearth of materials available for youth in high-risk urban communities. This article reports on secondary analyses of market research databases and other sources, as well as on primary research (e.g., focus groups) conducted with youth in high-risk urban communities. This research is exploratory in nature and is limited by the fact that the qualitative findings cannot be generalized to the overall population of high-risk youth. While the focus groups included youth from various races/ethnicities and geographical areas, CDC is aware that the study samples were not representative of the entire parent population in the United States. This exploratory research was conducted to inform the development of a communication campaign designed to reach youth in high-risk urban communities. Key findings for high-risk youth are provided across a variety of constructs including demographics, media and technology usage and impact, daily life and time spent in relationships, peer relationships, and attitudes and approaches to relationships. Implications for reaching this audience are discussed. These include reaching youth in urban settings and using approaches that focus on their peers, the Internet, cell phones, television, and music venues. Communication approaches that utilize print media or organized sports and other youth-oriented clubs and groups as channels to reach high-risk youth may be less impactful.
Conservative Management of Iatrogenic Esophageal Perforation During Neonatal Cardiac Surgery
Authors: Sasaki Takashi , Culham Gordon , Gandhi Sanjiv K.
Published: October 2012

Esophageal perforation is a rare, but life threatening, entity in children. The most common iatrogenic causes include nasogastric tube insertion, stricture dilation, or endotracheal intubation. Recently, transesophageal echocardiography (TEE) has been increasingly used in pediatric cardiac surgery to assess cardiac function and structural abnormalities. The safety of TEE in children is still controversial and complications such as airway obstruction, hemodynamic compromise, and esophageal injury have been reported. We recently experienced a case of esophageal perforation caused by TEE probe insertion during neonatal cardiac surgery; two weeks of conservative management resulted in complete resolution of the injury.
Using the UNOS/SRTR and PHTS Databases to Improve Quality in Pediatric Cardiac Transplantation
Authors: Davies Ryan R. , Pizarro Christian
Published: October 2012

Data collection and dissemination have been a part of the US transplant experience since its earliest days. As part of this process, the United Network for Organ Sharing (UNOS) has provided open access to its data. In addition, multiinstitutional groups such as the Pediatric Heart Transplant Study (PHTS) have collected data of particular interest to pediatric and congenital transplants. This wealth of data enables quality improvement along several pathways including individual program assessment and improvement and development of both structure and process measures for ongoing improvement. Extensive literature exists utilizing these data, but must be read critically, recognizing the limitations presented by missing variables (whether uncollected or collected but left blank), reproducibility, and small sample sizes among pediatric patients. However, despite these limitations, opportunity continues to exist to apply these data sets to ongoing questions of quality and optimize organ allocation and long-term survival among pediatric patients with heart failure.
Double Aortic Arch and Aortopulmonary Window
Authors: Caspi Joseph , Pettitt Timothy W. , Ascuito Robert J.
Published: October 2012

We describe a rare association of aortopulmonary window and double aortic arch in a 1.7-kg newborn who presented with severe respiratory distress. A staged surgical approach was used because of the size of the patient and significant comorbidity. This approach resulted in excellent outcome.
The Effects of Multiple Doses of Glucocorticoids on the Inflammatory Response to Cardiopulmonary Bypass in Children
Authors: Bronicki Ronald A. , Checchia Paul A. , Stuart-Killion Regan B. , Dixon David J. , Backer Carl L.
Published: October 2012

Background: We previously demonstrated that a dose of glucocorticoids (GCs) administered prior to cardiopulmonary bypass (CPB) is effective at suppressing the inflammatory response to CPB and leads to an improved postoperative course. We evaluated whether an additional dose of GC administered eight hours prior to CPB would lead to further clinical benefit. Methods: We conducted a prospective study in which patients were randomized to receive placebo or GC eight hours prior to CPB, in addition to a dose of GC administered following induction of anesthesia. We measured serum inflammatory mediator levels and postoperative clinical parameters. Results: Thirty-one patients were included in the study. Eighteen patients received two doses of GC and 13 patients received a single does of GC. Complement C3a levels were significantly lower at 24 hours following surgery in those patients who received two doses of GC (3136 ± 1650 vs 1779 ± 1616 ng/mL, P = .04). There was no significant difference in tumor necrosis factor (TNF)-α or interleukin (IL)-6 levels at any time between groups. There was no significant difference in core body temperature or renal function (based on serum creatinine levels) between groups. There was no significant difference between groups in duration of mechanical ventilation (2.4 ± 1.5 vs 3.6 ± 3.7 days, two vs one dose, respectively, P = .33) or length of stay in the intensive care unit ([ICU]; 3.4 ± 1.4 vs 4.9 ± 3.6 days, 2 vs 1 dose, respectively, P = .15). Conclusion: While those patients who received two doses of GC prior to surgery had significantly less complement activation postoperatively, clinical outcomes did not differ between groups. We conclude that the practice of administering an additional dose of GC prior to CPB is not supported. However, a large randomized study is needed to conclusively discount the potential benefit of this strategy.
Perventricular Closure of Muscular Ventricular Septal Defects in Infants With Echocardiographic Guidance Only
Authors: Fouilloux Virginie , Bonello Béatrice , Gran Célia , Fraisse Alain , Macé Loïc , Kreitmann Bernard
Published: October 2012

Objective: To report our experience with perventricular closure of muscular (apical) ventricular septal defects (VSDs) in small infants, with echocardiographic guidance only, in a nonhybrid suite. Methods: Eight infants with nine large muscular (apical) VSDs underwent perventricular device closure in a nonhybrid operating room, with transesophageal and epicardial echocardiography guidance, at a mean age and weight of 3.07 (0.3-7.28) months and 3.7 (2.5-6.2) kg, respectively. Five patients had multiple VSDs. Four had associated cardiac defects. Results: Nine Amplatzer muscular VSD devices with a mean size of 10 (4-14) mm were deployed. Seven patients were discharged from the intensive care unit with a mean length of stay of 8.6 days. Four patients had minimal postprocedural residual shunt; no one had a residual shunt at six-month follow-up. Mid-term results are excellent. Conclusion: Perventricular closure is feasible under echocardiographic guidance only in small patients, even without hybrid suite. This may be a good approach for very symptomatic low-weight infants with apical VSD and may also be useful, in any center, at any time, and in any operating room, to treat an associated apical VSD, even unexpected.
Apical Left Ventriculotomy is Safe in Infants and Young Children Requiring Cardiac Surgery
Authors: Goldberg Steven P. , Knott-Craig Christopher J. , Joshi Vijaya M. , Figueroa Mayte I. , Ballweg Jean A. , Chin Thomas K.
Published: October 2012

Objectives: Incisions in the left ventricle have previously been associated with increased mortality and morbidity, particularly in infants. In order to determine whether this assumption is still true in the current era, we reviewed our recent experience with apical left ventriculotomy in neonates and infants. Methods: The records of five consecutive patients requiring a left ventriculotomy between 2007 and 2010 were reviewed. Weight and age ranged from 2.6 to 16 kilograms and 5 days to 2 years. The diagnoses were three multiple ventricular septal defects, one rhabdomyoma, and one apical aneurysm. The primary end point was left ventricular ejection fraction, with other end points being intensive care unit length of stay, time to extubation, inotrope requirement, arrhythmias, and mitral valve function. Results: There were no early or late deaths. Although lower than their preoperative values, early postoperative ejection fractions were greater than 50% in all patients. Two patients required no inotropes, and 3 required only minimal support. Hospital length of stay was 9 ± 7 days for multiple ventricular septal defect patients, with intensive care unit stays of 2 to 5 days. There were no postoperative arrhythmias requiring pharmacological therapy, and one patient had a significant reduction in mitral insufficiency postoperatively. Conclusions: Based on our experience, we believe that an apical left ventriculotomy does not significantly impair left ventricular function even in small infants, and is not associated with significant morbidity, based on short-term follow-up. Although the long-term effects are still unknown, early results suggest that a left ventriculotomy may safely be used when alternative approaches are inadequate for complex cardiac defects.
Management of Pulmonary Arterial Supply Dependent on a Coronary Arterial Fistula in a Patient With Tetralogy of Fallot With Pulmonary Atresia
Authors: Garg Pankaj , Talwar Sachin , Kothari Shyam Sundar , Rajashekar Palleti , Gulati Gurpreet Singh , Anderson Robert H. , Airan Balram
Published: October 2012

In this report, we describe the surgical management of a patient with tetralogy of Fallot and pulmonary atresia, in whom a fistula from the anterior interventricular coronary artery was the predominant source of arterial supply to both the lungs.
Successful Simultaneous Correction of Complex Congenital Tracheal Stenosis and Tetralogy of Fallot
Authors: Arcieri Luigi , Moschetti Riccardo , Assanta Nadia , Cantinotti Massimiliano , Baggi Roberto , Mirabile Lorenzo , Murzi Bruno
Published: October 2012

Congenital tracheal stenosis is frequently associated with heart malformations. Simultaneous correction of both anomalies has been advocated by several authors. We describe our experience with a premature neonate with congenital tracheal stenosis and tetralogy of Fallot. The anomalies were corrected during the same surgical procedure with the aid of extracorporeal circulation. The implications of the operative and postoperative courses, concerning both the cardiac anomaly and the tracheal anomaly are discussed.
Surgical Repair of Tetralogy of Fallot at Age 83
Authors: Hussain Fareeda Taher Nazer , Grogan Martha , Dearani Joseph A.
Published: October 2012

Tetralogy of Fallot (TOF) is a cyanotic congenital heart defect typically diagnosed in infancy and treated with early surgical correction. We report a patient with TOF diagnosed at age 78. Successful surgical repair was performed at age 83, the oldest reported age of surgical correction of this condition. Despite a complicated surgical and postoperative course, the patient is doing well almost four years later.
Surgical Placement of Permanent Epicardial Pacing Systems in Very Low-Birth Weight Premature Neonates
Authors: Shepard Charles W. , Kochilas Lazaros , Vinocur Jeffrey M. , Bryant Roosevelt , Harvey Brian A. , Bradley Scott , DeCampli William , Louis James D. St.
Published: October 2012

Few studies have characterized the surgical outcomes following epicardial pacemaker placement in very low-birth weight infants with congenital complete heart block. This study was undertaken to review the surgical experience with this patient population based on data from a large multi-institutional registry. Methods: The Pediatric Cardiac Care Consortium (PCCC) multi-institutional database was retrospectively reviewed to identify premature, low-birth weight neonates that underwent surgical placement of an epicardial pacing system for heart block. We reviewed 179 patients with birth weights less than 1.5 kg that underwent a major operative procedure. Of these, 10 patients underwent surgical placement of an epicardial pacing system for heart block. Patients had heart block in otherwise structurally normal hearts (n = 6) or heart block associated with complex structural congenital cardiac anomalies (n = 4). Results: There were no deaths directly related to the surgical placement of the epicardial pacing system. There were no immediate complications with either lead or generator placement. One generator pocket was revised three months following placement. Survival to discharge was 60%. The four deaths occurred at a mean of 11 days (range 1-45 days) following the procedure. Conclusions: Neonates born with prematurity and congenital heart block represent a challenging subset of patients with significant mortality. Generator pocket breakdown and infection have been considered barriers to optimal short- and long-term outcomes. Among cases in the PCCC, there were no deaths or major complications that could be attributed to permanent epicardial pacemaker placement. These data suggest that an aggressive surgical strategy may be justified.
Bleeding and Thrombotic Emergencies in Pediatric Cardiac Intensive Care
Authors: Giglia Therese M. , DiNardo James , Ghanayem Nancy S. , Ichord Rebecca , Niebler Robert A. , Odegard Kirsten C. , Massicotte M. Patricia , Yates Andrew R. , Laussen Peter C. , Tweddell James S.
Published: October 2012

Children in the cardiac intensive care unit (CICU) with congenital or acquired heart disease are at risk for hematologic complications, both hemorrhage and thrombosis. The overall incidence of hematologic complications in the CICU is unknown, but risk factors and target groups have been identified where the essential physiologic balance between bleeding and clotting has been disrupted. Although the best management of life-threatening bleeding and clotting is prevention, the cardiac intensivist is often faced with managing life-threatening hematologic events involving patients from within the unit or those who present from outside. Part I of this review deals with the propensity of children with congenital and acquired heart disease to complications of both bleeding and clotting, and includes discussions of perioperative bleeding, thromboses in single-ventricle patients, clotting of Blalock-Taussig shunts and thrombotic complications of mechanical valves. Part II deals with the subject of stroke in children with heart disease. Part III reviews monitoring the effectiveness of anticoagulation and thrombolysis in the CICU. Currently available diagnostics modalities, medications and management strategies are reviewed and future directions discussed.
Role of the Skp1 prolyl-hydroxylation/glycosylation pathway in oxygen dependent submerged development of Dictyostelium
Authors: Xu Yuechi , Wang Zhuo A , Green Rebekah S , West Christopher M
Published: October 2012

Background

Oxygen sensing is a near universal signaling modality that, in eukaryotes ranging from protists such as Dictyostelium and Toxoplasma to humans, involves a cytoplasmic prolyl 4-hydroxylase that utilizes oxygen and α-ketoglutarate as potentially rate-limiting substrates. A divergence between the animal and protist mechanisms is the enzymatic target: the animal transcriptional factor subunit hypoxia inducible factor-α whose hydroxylation results in its poly-ubiquitination and proteasomal degradation, and the protist E3SCFubiquitin ligase subunit Skp1 whose hydroxylation might control the stability of other proteins. In Dictyostelium, genetic studies show that hydroxylation of Skp1 by PhyA, and subsequent glycosylation of the hydroxyproline, is required for normal oxygen sensing during multicellular development at an air/water interface. Because it has been difficult to detect an effect of hypoxia on Skp1 hydroxylation itself, the role of Skp1 modification was investigated in a submerged model of Dictyostelium development dependent on atmospheric hyperoxia.

Results

In static isotropic conditions beneath 70-100% atmospheric oxygen, amoebae formed radially symmetrical cyst-like aggregates consisting of a core of spores and undifferentiated cells surrounded by a cortex of stalk cells. Analysis of mutants showed that cyst formation was inhibited by high Skp1 levels via a hydroxylation-dependent mechanism, and spore differentiation required core glycosylation of Skp1 by a mechanism that could be bypassed by excess Skp1. Failure of spores to differentiate at lower oxygen correlated qualitatively with reduced Skp1 hydroxylation.

Conclusion

We propose that, in the physiological range, oxygen or downstream metabolic effectors control the timing of developmental progression via activation of newly synthesized Skp1.
Evaluation of cell proliferation, apoptosis, and dna-repair genes as potential biomarkers for ethanol-induced cns alterations
Authors: Hicks Steven D , Lewis Lambert , Ritchie Julie , Burke Patrick , Abdul-Malak Ynesse , Adackapara Nyssa , Canfield Kelly , Shwarts Erik , Gentile Karen , Meszaros Zsuzsa Szombathyne , Middleton Frank A
Published: October 2012

Background

Alcohol use disorders (AUDs) lead to alterations in central nervous system (CNS) architecture along with impaired learning and memory. Previous work from our group and that of others suggests that one mechanism underlying these changes is alteration of cell proliferation, apoptosis, and DNA-repair in neural stem cells (NSCs) produced as a consequence of ethanol-induced effects on the expression of genes related to p53-signaling. This study tests the hypothesis that changes in the expression of p53-signaling genes represent biomarkers of ethanol abuse which can be identified in the peripheral blood of rat drinking models and human AUD subjects and posits that specific changes may be correlated with differences in neuropsychological measures and CNS structure.

Results

Remarkably, microarray analysis of 350 genes related to p53-signaling in peripheral blood leukocytes (PBLs) of binge-drinking rats revealed 190 genes that were significantly altered after correcting for multiple testing. Moreover, 40 of these genes overlapped with those that we had previously observed to be changed in ethanol-exposed mouse NSCs. Expression changes in nine of these genes were tested for independent confirmation by a custom QuantiGene Plex (QGP) assay for a subset of p53-signaling genes, where a consistent trend for decreased expression of mitosis-related genes was observed. One mitosis-related gene (Pttg1) was also changed in human lymphoblasts cultured with ethanol. In PBLs of human AUD subjects seven p53-signaling genes were changed compared with non-drinking controls. Correlation and principal components analysis were then used to identify significant relationships between the expression of these seven genes and a set of medical, demographic, neuropsychological and neuroimaging measures that distinguished AUD and control subjects. Two genes (Ercc1 and Mcm5) showed a highly significant correlation with AUD-induced decreases in the volume of the left parietal supramarginal gyrus and neuropsychological measures.

Conclusions

These results demonstrate that alcohol-induced changes in genes related to proliferation, apoptosis, and DNA-repair are observable in the peripheral blood and may serve as a useful biomarker for CNS structural damage and functional performance deficits in human AUD subjects.
The HubBLe trial: haemorrhoidal artery ligation (HAL) versus rubber band ligation (RBL) for haemorrhoids
Authors: Tiernan Jim , Hind Daniel , Watson Angus , Wailoo Allan J , Bradburn Michael , Shephard Neil , Biggs Katie , Brown Steven
Published: October 2012

Background

Haemorrhoids (piles) are a very common condition seen in surgical clinics. After exclusion of more sinister causes of haemorrhoidal symptoms (rectal bleeding, perianal irritation and prolapse), the best option for treatment depends upon persistence and severity of the symptoms. Minor symptoms often respond to conservative treatment such as dietary fibre and reassurance. For more severe symptoms treatment such as rubber band ligation may be therapeutic and is a very commonly performed procedure in the surgical outpatient setting. Surgery is usually reserved for those who have more severe symptoms, as well as those who do not respond to non-operative therapy; surgical techniques include haemorrhoidectomy and haemorrhoidopexy. More recently, haemorrhoidal artery ligation has been introduced as a minimally invasive, non destructive surgical option.

There are substantial data in the literature concerning efficacy and safety of 'rubber band ligation including multiple comparisons with other interventions, though there are no studies comparing it to haemorrhoidal artery ligation. A recent overview has been carried out by the National Institute for Health and Clinical Excellence which concludes that current evidence shows haemorrhoidal artery ligation to be a safe alternative to haemorrhoidectomy and haemorrhoidopexy though it also highlights the lack of good quality data as evidence for the advantages of the technique.

Methods/design

The aim of this study is to establish the clinical effectiveness and cost effectiveness of haemorrhoidal artery ligation compared with conventional rubber band ligation in the treatment of people with symptomatic second or third degree (Grade II or Grade III) haemorrhoids.

Design: A multi-centre, parallel group randomised controlled trial.

Outcomes: The primary outcome is patient-reported symptom recurrence twelve months following the intervention. Secondary outcome measures relate to symptoms, complications, health resource use, health related quality of life and cost effectiveness following the intervention.

Participants: 350 patients with grade II or grade III haemorrhoids will be recruited in surgical departments in up to 14 NHS hospitals.

Randomisation: A multi-centre, parallel group randomised controlled trial. Block randomisation by centre will be used, with 175 participants randomised to each group.

Discussion

The results of the research will help inform future practice for the treatment of grade II and III haemorrhoids.

Trial Registration

ISRCTN41394716
A case–control study on the effect of Apolipoprotein E genotypes on gastric cancer risk and progression
Authors: De Feo Emma , Simone Benedetto , Persiani Roberto , Cananzi Ferdinando , Biondi Alberto , Arzani Dario , Amore Rosarita , D’Ugo Domenico , Ricciardi Gualtiero , Boccia Stefania
Published: October 2012

Background

Apolipoprotein E (ApoE) is a multifunctional protein playing both a key role in the metabolism of cholesterol and triglycerides, and in tissue repair and inflammation. The ApoE gene (19q13.2) has three major isoforms encoded by ε2, ε3 and ε4 alleles with the ε4 allele associated with hypercholesterolemia and the ε2 allele with the opposite effect. An inverse relationship between cholesterol levels and gastric cancer (GC) has been previously reported, although the relationship between apoE genotypes and GC has not been explored so far.

Methods

One hundred and fifty-six gastric cancer cases and 444 hospital controls were genotyped for apoE polymorphism (ε2, ε3, ε4 alleles). The relationship between GC and putative risk factors was measured using the adjusted odds ratios (ORs) and their 95% confidence intervals (CIs) from logistic regression analysis. A gene-environment interaction analysis was performed. The effect of the apoE genotypes on survival from GC was explored by a Kaplan–Meier analysis and Cox proportional hazard regression model.

Results

Subjects carrying at least one apoE ε2 allele have a significant 60% decrease of GC risk (OR=0.40, 95% CI: 0.19 – 0.84) compared with ε3 homozygotes. No significant interaction emerged between the ε4 or ε2 allele and environmental exposures, nor ε2 or ε4 alleles affected the median survival times, even after correcting for age, gender and stadium.

Conclusions

Our study reports for the first time a protective effect of the ε2 allele against GC, that might be partly attributed to the higher antioxidant properties of ε2 compared with the ε3 or ε4 alleles. Given the study’s sample size, further studies are required to confirm our findings.
“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”
Authors: Spath Katharina , Heinl Stefan , Grabherr Reingard
Published: October 2012

Background

Lactic acid bacteria (LAB) play an important role in agricultural as well as industrial biotechnology. Development of improved LAB strains using e.g. library approaches is often limited by low transformation efficiencies wherefore one reason could be differences in the DNA methylation patterns between the Escherichia coli intermediate host for plasmid amplification and the final LAB host. In the present study, we examined the influence of DNA methylation on transformation efficiency in LAB and developed a direct cloning approach for Lactobacillus plantarum CD033. Therefore, we propagated plasmid pCD256 in E. coli strains with different dam/dcm-methylation properties. The obtained plasmid DNA was purified and transformed into three different L. plantarum strains and a selection of other LAB species.

Results

Best transformation efficiencies were obtained using the strain L. plantarum CD033 and non-methylated plasmid DNA. Thereby we achieved transformation efficiencies of ~ 109 colony forming units/μg DNA in L. plantarum CD033 which is in the range of transformation efficiencies reached with E. coli. Based on these results, we directly transformed recombinant expression vectors received from PCR/ligation reactions into L. plantarum CD033, omitting plasmid amplification in E. coli. Also this approach was successful and yielded a sufficient number of recombinant clones.

Conclusions

Transformation efficiency of L. plantarum CD033 was drastically increased when non-methylated plasmid DNA was used, providing the possibility to generate expression libraries in this organism. A direct cloning approach, whereby ligated PCR-products where successfully transformed directly into L. plantarum CD033, obviates the construction of shuttle vectors containing E. coli-specific sequences, as e.g. a ColEI origin of replication, and makes amplification of these vectors in E. coli obsolete. Thus, plasmid constructs become much smaller and occasional structural instability or mutagenesis during E. coli propagation is excluded. The results of our study provide new genetic tools for L. plantarum which will allow fast, forward and systems based genetic engineering of this species.
Mechanical strain promotes osteoblast ECM formation and improves its osteoinductive potential
Authors: Guo Yong , Zhang Chun-qiu , Zeng Qiang-cheng , Li Rui-xin , Liu Lu , Hao Qin-xin , Shi Cai-hong , Zhang Xi-zheng , Yan Yu-xian
Published: October 2012

Background

The extracellular matrix (ECM) provides a supportive microenvironment for cells, which is suitable as a tissue engineering scaffold. Mechanical stimulus plays a significant role in the fate of osteoblast, suggesting that it regulates ECM formation. Therefore, we investigated the influence of mechanical stimulus on ECM formation and bioactivity.

Methods

Mouse osteoblastic MC3T3-E1 cells were cultured in cell culture dishes and stimulated with mechanical tensile strain. After removing the cells, the ECMs coated on dishes were prepared. The ECM protein and calcium were assayed and MC3T3-E1 cells were re-seeded on the ECM-coated dishes to assess osteoinductive potential of the ECM.

Results

The cyclic tensile strain increased collagen, bone morphogenetic protein 2 (BMP-2), BMP-4, and calcium levels in the ECM. Compared with the ECM produced by unstrained osteoblasts, those of mechanically stimulated osteoblasts promoted alkaline phosphatase activity, elevated BMP-2 and osteopontin levels and mRNA levels of runt-related transcriptional factor 2 (Runx2) and osteocalcin (OCN), and increased secreted calcium of the re-seeded MC3T3-E1 cells.

Conclusion

Mechanical strain promoted ECM production of osteoblasts in vitro, increased BMP-2/4 levels, and improved osteoinductive potential of the ECM. This study provided a novel method to enhance bioactivity of bone ECM in vitro via mechanical strain to osteoblasts.
Optimal surgical extent of lateral and central neck dissection for papillary thyroid carcinoma located in one lobe with clinical lateral lymph node metastasis
Authors: Keum Hyo Sub , Ji Yong Bae , Kim Jong Min , Jeong Jin Hyeok , Choi Woong Hwan , Ahn You Hern , Tae Kyung
Published: October 2012

Background

The indications and extent of cervical lymph node dissection in papillary thyroid carcinoma (PTC) are still being debated. The aim of this study was to analyze the patterns of cervical lymph node metastasis in the lateral and central compartment and related factors and suggest the optimal extent of lateral and central neck dissection for PTC patients with clinical lateral lymph node metastasis.

Methods

We retrospectively analyzed 72 patients with unilateral PTC who underwent therapeutic lateral neck dissections with concomitant total thyroidectomy and central neck dissection between January 2001 and December 2009.

Results

The 72 patients underwent 79 sides of therapeutic lateral neck dissection. The most frequent metastatic level in the ipsilateral lateral compartment was level IV (75.0%), followed by level III (69.4%), level II (56.9%) and level V (20.8%). Multiple level metastases were common (77.8%) and were correlated with tumor size (≥ 10 mm). The central compartment lymph node metastasis rate was 87.5%, including 26.4% of contralateral central compartment metastases.

Conclusion

In PTC patients with clinical lateral lymph node metastasis, the optimal extent of lateral and central neck dissection should include levels II, III, IV and V as well as the bilateral central compartment.
Rescue from acute neuroinflammation by pharmacological chemokine-mediated deviation of leukocytes
Authors: Berghmans Nele , Heremans Hubertine , Li Sandra , Martens Erik , Matthys Patrick , Sorokin Lydia , Van Damme Jo , Opdenakker Ghislain
Published: October 2012

Background

Neutrophil influx is an important sign of hyperacute neuroinflammation, whereas the entry of activated lymphocytes into the brain parenchyma is a hallmark of chronic inflammatory processes, as observed in multiple sclerosis (MS) and its animal models of experimental autoimmune encephalomyelitis (EAE). Clinically approved or experimental therapies for neuroinflammation act by blocking leukocyte penetration of the blood brain barrier. However, in view of unsatisfactory results and severe side effects, complementary therapies are needed. We have examined the effect of chlorite-oxidized oxyamylose (COAM), a potent antiviral polycarboxylic acid on EAE.

Methods

EAE was induced in SJL/J mice by immunization with spinal cord homogenate (SCH) or in IFN-γ-deficient BALB/c (KO) mice with myelin oligodendrocyte glycoprotein peptide (MOG35-55). Mice were treated intraperitoneally (i.p.) with COAM or saline at different time points after immunization. Clinical disease and histopathology were compared between both groups. IFN expression was analyzed in COAM-treated MEF cell cultures and in sera and peritoneal fluids of COAM-treated animals by quantitative PCR, ELISA and a bioassay on L929 cells. Populations of immune cell subsets in the periphery and the central nervous system (CNS) were quantified at different stages of disease development by flow cytometry and differential cell count analysis. Expression levels of selected chemokine genes in the CNS were determined by quantitative PCR.

Results

We discovered that COAM (2 mg i.p. per mouse on days 0 and 7) protects significantly against hyperacute SCH-induced EAE in SJL/J mice and MOG35-55-induced EAE in IFN-γ KO mice. COAM deviated leukocyte trafficking from the CNS into the periphery. In the CNS, COAM reduced four-fold the expression levels of the neutrophil CXC chemokines KC/CXCL1 and MIP-2/CXCL2. Whereas the effects of COAM on circulating blood and splenic leukocytes were limited, significant alterations were observed at the COAM injection site.

Conclusions

These results demonstrate novel actions of COAM as an anti-inflammatory agent with beneficial effects on EAE through cell deviation. Sequestration of leukocytes in the non-CNS periphery or draining of leukocytes out of the CNS with the use of the chemokine system may thus complement existing treatment options for acute and chronic neuroinflammatory diseases.
Prevalence of dyslipidemia among HIV-infected patients using first-line highly active antiretroviral therapy in Southern Ethiopia: a cross-sectional comparative group study
Authors: Tadewos Agete , Addis Zelalem , Ambachew Henock , Banerjee Sandip
Published: October 2012

Background

Data on lipid profile abnormalities among patients receiving highly active antiretroviral treatment in Ethiopia are very limited. The aim of this study was to determine the prevalence of dyslipidemia and characteristics of lipid profiles among patients living with human immunodeficiency virus (HIV) using first-line highly active antiretroviral therapy (HAART) in Southern Ethiopia.

Methods

This cross sectional comparative group study was conducted between March and May 2012, and included 113 HIV infected patients treated for a minimum of one year with first-line HAART regimens that included Efavirenz and Nevirapine (HAART group) and others 113 who had never received HAART (pre-HAART group). Serum lipid profiles were determined after overnight fasting and dyslipidemia was assessed according to the United State National Cholesterol Education program-III guideline. For statistical analysis Chi-square, student’s t-test, and logistic regression were used using Statistical Package for Social Sciences (SPSS) Version 20.

Result

Ninety-three (82.3%) of HAART and 87 (76.9%) pre-HAART patients had at least one laboratory abnormality, which is compatible with a diagnosis of dyslipidemia. Total cholesterol ≥ 200 mg/dl occurred in 43.4% of HAART and 15.9% pre-HAART patients (p=<0.0001), whereas HDL-cholesterol below 40 mg/dl occurred in 43.4% and in 63.7% respectively, (p=0.002). The LDL-cholesterol ≥ 130 mg/dl occurred in 33.6% of HAART and 15% pre-HAART patients (p=0.001), while triglycerides ≥ 150 mg/dl occurred in 55.8% and 31.0% respectively, (p=0.001). Receiving of HAART was significantly and positively associated with raised total cholesterol, LDL-cholesterol, and triglycerides. The adjusted odds ratio (95% CI) of HAART-treated vs. pre-HAART was 3.80 (1.34-6.55) for total cholesterol ≥ 200 mg/dl; 2.64 (1.31-5.32) for LDL- cholesterol ≥ 130 mg/dl and 2.50 (1.41-4.42) for triglycerides ≥150 mg/dl.

Conclusion

Use of first-line antiretroviral therapy regimens that contain Efavirenz and Nevirapine were associated with raised total cholesterol, LDL-cholesterol, and triglycerides, an established atherogenic lipid profiles. Lipid profiles should be performed at baseline before commencement of antiretroviral therapy and then periodically through treatment follow-up to monitor any rising trends.
Infant feeding practices and breastfeeding duration in Japan: A review
Authors: Inoue Madoka , Binns Colin W , Otsuka Keiko , Jimba Masamine , Matsubara Manami
Published: October 2012

The Japanese health system places great emphasis on healthy development. However, the prevalence of Exclusive Breastfeeding at one month postpartum between 1980 and 2005 has remained unchanged, fluctuating between 42% and 49%. At the same time, the Any Breastfeeding prevalence has gradually increased from about 80% to 95%. In 2010, the latest national breastfeeding report showed that ‘exclusive’ and ‘any’ breastfeeding rates have improved. However, as the World Health Organization (WHO) definition of breastfeeding practices was not used in this study or in other national surveys, it is difficult to interpret these latest results. While the Japanese government has launched several promotion projects, there have been few studies and reviews of risk factors that influence breastfeeding duration. The objectives of this review were to summarise the factors that have influenced the duration of breastfeeding in Japan to provide information relevant to breastfeeding promotion programs. A search of electronic databases in Japanese and English was undertaken up to 2011. The inclusion criteria for this review were studies that focused on infant feeding practices and targeted Japanese mothers, fathers, or health professionals, but excluded mothers’ friends and peer groups. In total, 12 articles were selected for the final analysis. Smoking status, low birth weight of infants and maternal perceptions of insufficient breast milk supply were negative influences on breastfeeding duration, while support from husbands/partners is associated with continued breastfeeding. Some factors that have been found to be associated with breastfeeding in other countries, including maternal age, family income, maternal educational levels, and living with grandparents of infants have not been confirmed in Japan. While the national breastfeeding rates were higher than other countries of similar health status, inconsistent knowledge of breastfeeding benefits and inappropriate hospital practices remain in Japan may be associated with increased the use of infant formula and reduced breastfeeding duration. Most of the studies reviewed were cross-sectional in design, with only a limited number of cohort studies. Also many published studies used small sample sizes. Cohort studies of infant feeding practices with larger sample sizes are required to monitor trends in rates and risk factors for breastfeeding outcomes.
Identification of resistant carboxylesterase alleles in Culex pipiens complex via PCR-RFLP
Authors: Zhang Hanying , Meng Fengxia , Qiao Chuanling , Cui Feng
Published: October 2012

Background

Carboxylesterase overproduction is a frequently observed resistance mechanism of insects to organophosphate insecticides. As a major transmitter of human diseases, mosquitoes in theCulex pipienscomplex have evolved 13 carboxylesterase alleles (Ester) that confer organophosphate resistance. Six alleles,EsterB1,Ester2,Ester8,Ester9,EsterB10, andEster11, have been observed in field populations in China, sometimes co-existing in one population. To differentiate the carboxylesterase alleles found in these field populations, PCR-RFLP was designed for use in resistance monitoring.

Results

Based on the DNA sequences of resistant and nonresistant carboxylesterase alleles,Ester Balleles were first amplified with PCR-specific primers and then digested with the restriction enzymeDraI. In this step,Ester2andEster11were differentiated from the otherEsteralleles. When the otherEsterB alleles were digested with the restriction enzymeXbaI,EsterB1and the susceptibleC. p. pallens Esterwere screened out.Ester8andEster9were differentiated fromEsterB10and the susceptibleC. p. quinquefasciatusesterase allele, respectively, by amplifying and digesting theEster Aalleles with the restriction enzymeApaLI. The effectiveness of the custom-designed PCR-RFLP was verified in two field mosquito populations.

Conclusions

A PCR-RFLP based approach was developed to differentiate carboxylesterase alleles inCulex pipienscomplex mosquitoes. These processes may be useful in monitoring the evolutionary dynamics of known carboxylesterase alleles as well as in the identification of new alleles in field populations.
Pattern of outsole shoe heel wear in infantry recruits
Authors: Finestone Aharon S , Petrov Kaloyan , Agar Gabriel , Honig Assaf , Tamir Eran , Milgrom Charles
Published: October 2012

Background

Excessive shoe heel abrasion is of concern to patients, parents and shoe manufacturers, but little scientific information is available. The purpose of this study was to describe the phenomenon in a group of infantry recruits performing similar physical activity, and search for biomechanical factors that might be related.

Methods

Seventy‐six subjects (median age 19) enrolled. Pre‐training parameters measured included height, weight, tibial length, foot arch height and foot progression angle. Digital plantar pressure maps were taken to calculate arch indexes. Shoe heel abrasion was assessed manually after 14 weeks of training with different‐sized clock transparencies and a calliper.

Results

Outsole abrasion was posterolateral, averaging 12 degrees on each shoe. The average heel volume that was eroded was almost 5 cm3. The angle of maximum wear was related to right foot progression angle (r = 0.27, p = 0.02). Recruits with lateral ankle sprains had higher angles of maximal abrasion (17° versus 10°, p = 0.26) and recruits with lateral heel abrasion had more lateral ankle sprains (14% versus 3%, p = 0.12).

Conclusion

While shoe heel wear affects many people, very little has been done to measure it. In this study in healthy subjects, we found the main abrasion to be posterolateral. This seems to be related to foot progression angle. It was not related to hindfoot valgus/varus or other factors related to subtalar joint motion. These findings do not warrant modification of subtalar joint motion in order to limit shoe heel abrasion.
Numerical analysis of airflow alteration in central airways following tracheobronchial stent placement
Authors: Ho Chien-Yi , Liao Hsiu-Mei , Tu Chih-Yen , Huang Chih-Yang , Shih Chuen-Ming , Su Min-Ying Lydia , Chen Jeon-Hor , Shih Tzu-Ching
Published: October 2012

The computational fluid dynamics method, which provides an estimation of the pressure drop in the airway before and after the stent implantation, is proposed in this study. This method is based on the finite volume model. The pressure field was solved by the Navier-Stokes equations. The proposed methodology was evaluated in seven health people (control group) and in fourteen patients who were assigned in two groups, in which one was tracheal stenosis and the other was bronchial stenosis. The results showed that the pressure drop after tracheal stent implantation became significantly smaller. For bronchial stent implantation cases, the airway resistance improved insignificantly.