Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development.

Chlamydia is a prevalent sexually transmitted disease that infects more than 100 million people worldwide. Although most individuals infected with Chlamydia trachomatis are initially asymptomatic, symptoms can arise if left undiagnosed. Long-term infection can result in debilitating conditions such as pelvic inflammatory disease, infertility, and blindness. Chlamydia infection, therefore, constitutes a significant public health threat, underscoring the need for a Chlamydia-specific vaccine. Chlamydia strains express a major outer-membrane protein (MOMP) that has been shown to be an effective vaccine antigen. However, approaches to produce a functional recombinant MOMP protein for vaccine development are limited by poor solubility, low yield, and protein misfolding. Here, we used an Escherichia coli-based cell-free system to express a MOMP protein from the mouse-specific species Chlamydia muridarum (MoPn-MOMP or mMOMP). The codon-optimized mMOMP gene was co-translated with Δ49apolipoprotein A1 (Δ49ApoA1), a truncated version of mouse ApoA1 in which the N-terminal 49 amino acids were removed. This co-translation process produced mMOMP supported within a telodendrimer nanolipoprotein particle (mMOMP-tNLP). The cell-free expressed mMOMP-tNLPs contain mMOMP multimers similar to the native MOMP protein. This cell-free process produced on average 1.5 mg of purified, water-soluble mMOMP-tNLP complex in a 1-ml cell-free reaction. The mMOMP-tNLP particle also accommodated the co-localization of CpG oligodeoxynucleotide 1826, a single-stranded synthetic DNA adjuvant, eliciting an enhanced humoral immune response in vaccinated mice. Using our mMOMP-tNLP formulation, we demonstrate a unique approach to solubilizing and administering membrane-bound proteins for future vaccine development. This method can be applied to other previously difficult-to-obtain antigens while maintaining full functionality and immunogenicity.

The role of clopidogrel in the management of ischemic heart disease.

PURPOSE OF REVIEW: Placebo-controlled randomized trials have demonstrated the efficacy of clopidogrel in combination with aspirin across a broad range of clinical presentations. Recent trials have addressed several remaining issues regarding clopidogrel therapy. RECENT FINDINGS: Three randomized trials examined the role of platelet function testing (PFT) in clopidogrel-treated patients. The results do not support the use of PFT to adjust clopidogrel dose after percutaneous coronary intervention (PCI), particularly in patients with stable angina or ischemia, in whom event rates are low irrespective of on-treatment reactivity. Doses greater than clopidogrel 150  mg daily are required to sufficiently overcome high reactivity in CYP2C19 loss-of-function (LOF) allele carriers. Clopidogrel response variability also influences the time to platelet recovery after drug discontinuation, and a proof-of-principle study supports the concept of using PFT for surgical timing. Unlike its efficacy in the setting of acute coronary syndrome (ACS) and PCI, prasugrel was not superior to clopidogrel in medically treated patients recovering from ACS. SUMMARY: Current data do not support routine PFT to guide antiplatelet therapy in patients undergoing nonurgent PCI. The role of PFT to optimize therapy in ACS patients remains unaddressed, and further study is needed to confirm its promise in guiding surgical timing in patients who have discontinued therapy. Clopidogrel remains an important therapeutic option for patients presenting after an ACS who did not undergo initial revascularization.

Long-term clinical outcomes of real-world experience using sirolimus-eluting stents in saphenous vein graft disease.

OBJECTIVE: To evaluate the long-term clinical outcomes of patients undergoing percutaneous coronary intervention for saphenous vein graft (SVG) disease. Specifically, we compared clinical endpoints of patients who received sirolimus-eluting stents (SES) versus bare-metal stents (BMS) for SVG disease. BACKGROUND: A recent small randomized-controlled trial (RCT) reported increased mortality with the use of SES in SVG disease. METHODS: We retrospectively identified patients who underwent SES placement for a SVG lesion(s) at our institutions over a 4-year period. The procedural and medical records were reviewed to identify predetermined clinical outcomes. RESULTS: 318 patients who underwent SES placement for a SVG lesion were identified. 7 patients were lost to follow-up. 141/311 patients (45%) received SES, while 170/311 (55%) received BMS. At a mean follow-up of 34 months, there was a reduction in target lesion revascularization (TLR) (7% vs. 14%, P = 0.07) without an increased risk of mortality (6% vs. 12%, P = 0.06) in patients who received SES compared to patients who received BMS. When compared to the recent RCT's SES patients at long-term follow-up, our SES patients had significantly less mortality; rates of myocardial infarction, TLR, target vessel revascularization, and major adverse cardiac events; and were more likely to be taking dual antiplatelet and statin medications. CONCLUSION: Our results support that SES used in SVG lesions result in a reduction in TLR without an increased risk of mortality, and therefore may be an equally safe and feasible technique for revascularization with excellent long-term clinical outcomes. These patients may benefit from prolonged dual antiplatelet and statin medication regimens.

The hidden curriculum: medical students' changing opinions toward the pharmaceutical industry.

BACKGROUND: Authorities suggest academic medical centers eliminate conflicts of interest. The authors evaluated medical students' opinions and knowledge of the pharmaceutical industry. METHOD: An anonymous 20-item questionnaire was administered to medical students from four different medical schools; 15 items addressed opinions, and five items were free-response knowledge questions. Results were analyzed by Fisher exact test. RESULTS: Authors received 667 responses from the schools. Sixty-five percent of clinical students believed accepting gifts was appropriate; 28% of preclinical students believed it was appropriate (P < .001). Knowledge was the same for clinical and preclinical students. CONCLUSIONS: Clinical students were more favorable toward receiving gifts than were preclinical students, yet there was no difference in their knowledge of the industry. Increased formal and informal education about the pharmaceutical industry is necessary during the clinical years.

Coffee: good, bad, or just fun? A critical review of coffee's effects on liver enzymes.

Coffee consumption is a regular part of daily life throughout the world. Research into the effects of coffee on human health is ongoing, but a recent study suggests that coffee and caffeine consumption can reduce the risk of elevated alanine aminotransferase activity in individuals at high risk for liver disease. This review will analyze the results of that study in light of the current literature.