Maternal high-fat diet associated with altered gene expression, DNA methylation, and obesity risk in mouse offspring.

We investigated maternal obesity in inbred SM/J mice by assigning females to a high-fat diet or a low-fat diet at weaning, mating them to low-fat-fed males, cross-fostering the offspring to low-fat-fed SM/J nurses at birth, and weaning the offspring onto a high-fat or low-fat diet. A maternal high-fat diet exacerbated obesity in the high-fat-fed daughters, causing them to weigh more, have more fat, and have higher serum levels of leptin as adults, accompanied by dozens of gene expression changes and thousands of DNA methylation changes in their livers and hearts. Maternal diet particularly affected genes involved in RNA processing, immune response, and mitochondria. Between one-quarter and one-third of differentially expressed genes contained a differentially methylated region associated with maternal diet. An offspring high-fat diet reduced overall variation in DNA methylation, increased body weight and organ weights, increased long bone lengths and weights, decreased insulin sensitivity, and changed the expression of 3,908 genes in the liver. Although the offspring were more affected by their own diet, their maternal diet had epigenetic effects lasting through adulthood, and in the daughters these effects were accompanied by phenotypic changes relevant to obesity and diabetes.

Microaggressions, Feelings of Difference, and Resilience Among Adopted Children with Sexual Minority Parents.

Limited research exists about the unique experiences and possible marginalization of children with sexual minority parents. From a larger longitudinal project of diverse adoptive families, we examined cross-sectional data using mixed methods from interviews with 49 adopted children (M age = 8 years; 47% female) in 27 two-father and 22 two-mother families. Using thematic analysis, we coded themes of awareness of difference, microaggressions, and resilience (i.e., coping and positive family conceptualizations). Children experienced "feeling different" and microaggressions from peers, but generally at a low to medium intensity and with neutral (not negative) emotion. More instances of resilience and positive family conceptualizations were reported than microaggressions or feelings of difference, suggesting that children develop positive perceptions of their family and navigate experiences of difference with resilience. Filling important gaps in the literature, we discuss implications of our results for practice and policy.

Outcome study of brief relational-cultural therapy in a women's mental health center.

The current study is the first to evaluate the effectiveness of a brief relational-cultural model of therapy in a community-based mental health center for women. The study was distinctive in its use of a hybrid model that employed elements of randomized control and naturalistic design. Results showed that the entire treatment group of 91 women improved significantly on all eight outcome measures. Therapeutic gains were maintained at 3- and 6-month follow-ups. The results lend support to the importance of including relational-cultural factors in the treatment of women. An adherence scale/manual was developed and implemented and will allow for replication.

The St. Leger total knee replacement: a 10-year clinical and radiological assessment.

The St. Leger total knee replacement (Zynergy Orthopaedics Ltd, Rotherham, UK) was developed as a cheaper alternative to similar implants of its time. Between October 1993 and June 1999, 144 St. Leger total knee replacements were implanted into 114 patients. Seventy-three patients (99 knees) were recalled for assessment (mean follow-up of 10.2 years). Eighteen patients had had their prostheses revised, 11 had died and 12 were lost to follow-up. Functional Score showed 90% poor results and the Objective Knee Score showed 31% poor results. Radiological assessment identified 12 arthroplasties that had failed and 58 that required close follow. Kaplan-Meier cumulative survivorship was 87% at 10 years. The St. Leger knee replacement did not perform as well as others of the same generation and was not worth the initial financial savings.

The many faces of the genetics contribution to temporomandibular joint disorder.

OBJECTIVES: Review the literature on candidate genes for temporomandibular joint disorder (TMD). Setting and Sample Population - Literature review. MATERIALS AND METHODS: Two basic approaches were used to obtain literature in any language regarding genes and TMD. First, Medline, Embase, and Science Citation Index databases were searched using the keywords 'temporomandibular joint disorder' and 'temporomandibular joint dysfunction' for studies published from 1966 to 2007. Then, the references list of the studies obtained in the database was also considered. RESULTS: Candidate genes for TMD include genes for individual variations in pain perception, gender and ethnicity, proinflammatory cytokines, female hormones, breakdown of extracellular matrix, and syndromic forms of TMD. CONCLUSION: Most of the studies on genetic variation contributing to TMD are approaching the disease mainly from an immune-inflammatory perspective. Recent investigations of the genetic variables which may predict identifiable levels of pain perception may uncover new approaches to our traditional treatment modalities for the chronic pain patient.

The endangered clinical teacher scholar: will this eliminate discovery from the dental school environment?

Each year, dental schools struggle to do more with less-a reality that is confounded by dental faculty shortages and retention issues reported over the past 15 years. In today's academic environment, faculty shortages suggest that the resulting smaller faculty population will be tasked with more responsibilities than those who had the benefit of working with a full complement of colleagues. Fewer clinical faculty will likely participate in scholarly activity-in some instances, negatively affecting their chances for promotion and tenure, and ultimately lessening their own job security and feeling of accomplishment and self-worth in the academic environment. New faculty are in need of a formal program endorsed by their administration-one which places them with colleagues experienced in scholarly activity. This program, to include a definitive reward structure for mentors, would likely foster the retention and promotion of faculty and encourage the development of future leaders of dental education. Without a definitive plan, the clinical scholar will become an endangered species, and the research innovations and discovery of our profession will fall short in addressing the needs of the public's oral and systemic health.

Optimum configuration of cannulated hip screws for the fixation of intracapsular hip fractures: a biomechanical study.

The radiographs of a sample of patients who had canulated hip screw fixation for intracapsular femoral neck fractures were reviewed in our region. There were six different types of configurations used in these fixations which are divided into two groups: (I) triangular configurations, consisting of two parallel screws with a third screw placed either superiorly, inferiorly, anteriorly or posteriorly; and (II) linear configurations with two or three screws in a vertical line. In our study, we tested the relative strength of each configuration in a laboratory setting using synthetic bone models. Statistical analysis, at 5% significance level, using two-way ANOVA and post-hoc test was carried out to test the differences of the results between the configurations.Our results clearly show that the triangular configurations had a higher peak load, higher ultimate load, less displacement and more energy absorption before failure than other configurations.

Left principal bronchus rupture in a cat.

A two-year-old neutered male domestic shorthaired cat was referred with respiratory compromise. Investigations included thoracic radiography and tracheal endoscopy. Findings were consistent with a traumatic avulsion injury to the left principal bronchus. Treatment involved the resection of the ruptured and stenosed left principal bronchus segment via a fifth right intercostal lateral thoracotomy. Once the damaged portion of bronchus had been removed, bronchial repair was achieved by end-to-end anastomosis. A full recovery was made and, at the time of writing (16 months postoperatively), the cat was clinically normal.

Codon optimization of gene fragments encoding Plasmodium falciparum merzoite proteins enhances DNA vaccine protein expression and immunogenicity in mice.

In contrast to conventional vaccines, DNA and other subunit vaccines exclusively utilize host cell molecules for transcription and translation of proteins. The adenine plus thymine content of Plasmodium falciparum gene sequences (approximately 80%) is much greater than that of Homo sapiens (approximately 59%); consequently, codon usage is markedly different. We hypothesized that modifying codon usage of P. falciparum genes encoded by DNA vaccines from that used by the parasite to those resembling mammalian codon usage would lead to increased P. falciparum protein expression in vitro in mouse cells and increased antibody responses in DNA-vaccinated mice. We synthesized gene fragments encoding the receptor-binding domain of the 175-kDa P. falciparum erythrocyte-binding protein (EBA-175 region II) and the 42-kDa C-terminal processed fragment of the P. falciparum merozoite surface protein 1 (MSP-1(42)) using the most frequently occurring codon in mammals to code for each amino acid, and inserted the synthetic genes in DNA vaccine plasmids. In in vitro transient-expression assays, plasmids containing codon-optimized synthetic gene fragments (pS plasmids) showed greater than fourfold increased protein expression in mouse cells compared to those containing native gene fragments (pN plasmids). In mice immunized with 0.5, 5.0, or 50 microg of the DNA plasmids, the dose of DNA required to induce equivalent antibody titers was 10- to 100-fold lower for pS than for pN plasmids. These data demonstrate that optimizing codon usage in DNA vaccines can improve protein expression and consequently the immunogenicity of gene fragments in DNA vaccines for organisms whose codon usage differs substantially from that of mammals.

A GCN4 variant with a C-terminal basic region binds to DNA with wild-type affinity.

Basic-region leucine zipper (bZip) proteins contain a bipartite DNA-binding motif consisting of a leucine zipper dimerization domain and a basic region that directly contacts DNA. In all naturally occurring bZip proteins, the basic region is positioned N-terminal to the leucine zipper. We have designed a series of model bZip peptides in which the basic region of the yeast transcriptional activator GCN4 is placed C-terminal to its leucine zipper. DNA-binding studies demonstrate that the optimal reverse GCN4 (rGCN4) peptide is able to bind specifically and with wild-type affinity to DNA despite this unnatural arrangement of the two subdomains. These results suggest that a thermodynamic basis for the observed N-terminal positioning of the basic region relative to the dimerization domain is unlikely.

Selection of a high-affinity DNA pool for a bZip protein with an out-of-phase alignment of the basic region relative to the leucine zipper.

bZip transcription factors contain two regions that are required for DNA binding: a leucine zipper dimerization domain and a highly charged basic region that directly contacts DNA. The spacing between these subdomains is strictly conserved, and changes in this spacing result in a loss of function. Using an in vitro selection strategy, we have investigated the ability of a bZip protein with incorrect spacing between these two regions to bind specifically to DNA. Surprisingly, we find that although such a protein does not bind to its predicted site, it is possible to isolate a pool of DNAs that bind with very similar affinity to that of GCN4 for its optimum DNA site.

The design of antiparallel coiled coils.

Recent structural studies have highlighted the importance of antiparallel coiled coils in nature. In addition, well-behaved, model antiparallel coiled coils have been designed and used for the reassembly of protein fragments and for the study of the energetic contributions of various interactions to helix orientation specificity. Finally, high-resolution structural data are available for designed helical bundles, allowing an evaluation of the success of state-of-the-art protein design efforts.

Subvastus approach for total knee arthroplasty: a prospective, randomized, and observer-blinded trial.

A prospective, randomized, and blinded trial was conducted with 89 consecutive primary knee arthroplasties comparing standard medial parapatellar arthrotomy with the subvastus approach. All patients received the same prosthesis (Insall-Burstein II) inserted by one surgeon using an identical technique with the only difference being the approach. The parapatellar approach (group I) was used in 43 knees, and in the remaining 46 knees the subvastus approach (group II) was used. Assessment revealed significantly earlier return of straight-leg raise (3.2 days vs 5.8 days, P <.001), lower consumption of opiates in the first week (78 mg vs 102 mg, P <.001), less blood loss (527 mL vs 748 mL, P <.001), and greater knee flexion at 1 week (78 degrees vs 55 degrees, P <.001) in group II (subvastus approach). The subvastus approach offers early advantages over the standard parapatellar arthrotomy. It preserves the integrity of the vastus medialis and peripatellar plexus of vessels. We advise its wider use in primary total knee arthroplasty.

Developing and using preparatory information for women undergoing radiation therapy for cervical or uterine cancer.

PURPOSE/OBJECTIVES: To describe symptoms associated with radiation therapy necessary to develop preparatory concrete, objective information for women with cervical or uterine cancer. DESIGN: Prospective, descriptive. SETTING: University-affiliated radiation oncology department in the mid-south. SAMPLE: 49 of 52 women undergoing external beam radiation and low-dose rate brachytherapy for cervical or uterine cancer. Stage of disease ranged from I-IIIA. Mean age of subjects was 54 years; mean years of education was 11. METHODS: Investigator-developed symptom checklist based on prior research. MAIN RESEARCH VARIABLES: Symptom reports. FINDINGS: At least 40% of the women reported difficulty sleeping, fatigue, diarrhea, anorexia, nausea, urinary frequency, dysuria, vaginal discharge, and perineal irritation. Incidence and timing of symptoms varied by operative status and brachytherapy timing. CONCLUSIONS: Findings confirm and clarify the symptoms women associate with radiation treatment for cervical or uterine cancer. IMPLICATIONS FOR NURSING PRACTICE: Assessing research findings for relevance to the care of patients in specific settings is necessary. These findings are appropriate for developing preparatory information for women undergoing external beam radiation and low-dose rate brachytherapy.

Evaluation of the energetic contribution of interhelical Coulombic interactions for coiled coil helix orientation specificity.

Coiled coils are formed by two or more alpha-helices that align in a parallel or an antiparallel relative orientation. The factors that determine a preference for a given relative helix orientation are incompletely understood. The helix orientation preference for the designed coiled coil, Acid-a1-Base-a1, was measured previously. This model system therefore provides a means for the experimental determination of the energetic contribution of a variety of interactions to helix orientation specificity. The antiparallel preference for Acid-a1-Base-a1 is imparted by a single buried polar interaction. Interhelical Coulombic interactions between residues at the e and g positions have been proposed to influence helix orientation preference. In the Acid-a1-Base-a1 heterodimer, potentially attractive Coulombic interactions are expected in both orientations. To determine the energetic consequences of Coulombic interactions for helix orientation preference, we have positioned a single charged residue in each peptide such that exclusively favorable interhelical Coulombic interactions can occur only in the parallel orientation. In contrast, two potentially repulsive interactions are expected in the antiparallel orientation. Because the buried polar interaction can occur only in the antiparallel orientation, interhelical Coulombic interactions favor the parallel orientation and the potential to form a buried polar interaction favors the antiparallel orientation. We find no clear preference for an antiparallel orientation in the resulting heterodimer, Acid-Ke-Base-Eg, suggesting that interhelical Coulombic interactions and a buried polar interaction are of approximately equal importance for helix orientation specificity. Stability measurements indicate that maintenance of all favorable electrostatic interactions and/or avoidance of two potentially repulsive interactions contributes approximately 2.1 kcal/mol to helix orientation preference.

GCN4 binds with high affinity to DNA sequences containing a single consensus half-site.

bZip proteins contain a bipartite DNA-binding motif consisting of a "leucine zipper" dimerization domain and a highly charged "basic region" that directly contacts DNA. These transcription factors form dimeric complexes with each monomer recognizing half of a symmetric or nearly symmetric DNA site. We have found that the bZip protein GCN4 can also bind with high affinity to DNA sites containing only a single GCN4 consensus half-site. Because several recent lines of evidence have suggested a role for monomeric DNA binding by bZip proteins, we investigated the structure of the GCN4.half-site complex. Quantitative DNA binding and affinity cleaving studies support a model in which GCN4 binds as a dimer, with one monomer making specific contacts to the consensus half-site and the other monomer forming nonspecific contacts that are nonetheless important for binding affinity. We also examined the folding transition induced in the basic regions of this complex upon binding DNA. Circular dichroism (CD) studies demonstrate that the basic regions of both monomers are helical, suggesting that a protein folding transition may be required for both specific and nonspecific DNA binding by GCN4.

A buried polar interaction can direct the relative orientation of helices in a coiled coil.

Coiled coils consist of bundles of two or more alpha-helices that are aligned in a parallel or an antiparallel relative orientation. The designed peptides, Acid-p1 and Base-p1, associate in solution to form a parallel, heterodimeric two-stranded coiled coil [O'Shea, E. K., Lumb, K. J., and Kim, P. S. (1993) Curr. Biol. 3, 658]. The buried interface of this complex is formed by hydrophobic Leu residues, with the exception of an Asn residue from each strand that is positioned to engage in a buried polar interaction. Substitution of these buried Asn residues by Leu residues results in a loss of structural uniqueness, as evidenced by a lack of a particular helix orientation in the Acid-Base coiled-coil complex [Lumb, K. J., and Kim, P. S. (1995) Biochemistry 34, 8642]. Here, we alter the positions of the Asn residues in the Acid and Base peptides such that a buried polar interaction is only expected to occur when the helices are in an antiparallel orientation. The resulting peptides, Acid-a1 and Base-a1, associate to form a helical heterodimer, as shown by circular dichroism (CD) and equilibrium sedimentation centrifugation. The helix orientation preference has been measured using covalently linked, disulfide-containing heterodimers in which the constituent peptides are constrained to interact in either a parallel or an antiparallel orientation. Although both the parallel and antiparallel heterodimers form stable, helical structures, the antiparallel heterodimer is the predominant species at equilibrium when the heterodimers are allowed to undergo thiol-disulfide exchange. In addition, the antiparallel heterodimer is more stable to chemical denaturation than the parallel counterpart by approximately 2.3 kcal/mol. These results demonstrate that a single buried polar interaction in the interface between the helices of a coiled coil is sufficient to determine the relative orientation of its constituent helices.

Randomized placebo-controlled trial of local anaesthetic infusion in day-case inguinal hernia repair.

BACKGROUND: This study investigated the efficacy of local anaesthetic wound perfusion following day-case inguinal hernia repair. METHODS: Seventy-two patients entered a randomized controlled trial with three patient groups: group 1, pump containing bupivacaine; group 2, pump containing normal saline; and group 3, control group without a pump. All patients had a Lichtenstein hernia repair together with ilioinguinal and iliohypogastric nerve blocks and were prescribed oral analgesia. Postoperative pain was assessed over 5 days using a visual analogue scale. RESULTS: Patients who had a local anaesthetic infusion had significantly less pain than either the placebo or control groups. This was greatest during the first 48 h (day 1, P = 0.028 and 0.011 respectively; day 2, P = 0.012 and 0.037 respectively). CONCLUSION: A portable infusion pump for the delivery of local anaesthetic reduced pain after day-case inguinal hernia repair.