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Antisynthetase syndrome is a rare multisystem autoimmune disorder which clinically manifests with myositis, arthritis, interstitial lung disease, Raynaud phenomenon, and skin hyperkeratosis. Lung involvement represents the most severe form of disease and has rarely been reported in pregnancy. We present the case of a 22-year-old woman with antisynthetase syndrome and severe restrictive pulmonary disease who experienced a successful pregnancy and delivery. We discuss anesthetic considerations and highlight the importance of a multidisciplinary team approach in caring for parturients with multifactorial medical conditions. [ABSTRACT FROM AUTHOR]

Background De Barsy syndrome is a rare, autosomal recessive syndrome characterized by cutis laxa, progeroid appearance, ophthalmic opacification, skeletal malformations, growth delays, and intellectual disability. Aims The aim of this case series is to identify the anesthetic considerations in the clinical management of patients with de Barsy syndrome. Methods A retrospective case review from 1968 to 2016 was performed at a single tertiary medical center to identify patients with de Barsy syndrome who underwent anesthesia for diagnostic and surgical procedures. We collected and analyzed the perioperative records and following data: age, sex, American Society of Anesthesiologists physical status, relevant comorbidities, surgical procedures, anesthesia management, and observed complications. Results Three patients underwent 64 unique anesthetics for a diverse collection of diagnostic and surgical procedures. An array of anesthetics and techniques were successfully used. Observations of the perioperative period found 7 episodes of intraoperative hyperthermia (>38.3°), a single difficult airway requiring fiberoptic bronchoscopic-guided intubation, and repeatedly difficult intravenous access. Conclusion This expanded case series suggests that providers caring for patients with de Barsy syndrome should be aware of potential challenges with airway management, vascular access, and temperature monitoring. [ABSTRACT FROM AUTHOR]

Background. Ipsilateral phrenic nerve blockade is a common adverse event after an interscalene brachial plexus block, which can result in respiratory deterioration in patients with preexisting pulmonary conditions. Diaphragm-sparing nerve block techniques are continuing to evolve, with the intention of providing satisfactory postoperative analgesia while minimizing hemidiaphragmatic paralysis after shoulder surgery. Case Report. We report the successful application of a combined ultrasound-guided infraclavicular brachial plexus block and suprascapular nerve block in a patient with a complicated pulmonary history undergoing a total shoulder replacement. Conclusion. This case report briefly reviews the important innervations to the shoulder joint and examines the utility of the infraclavicular brachial plexus block for postoperative pain management. [ABSTRACT FROM AUTHOR]

Study Objective: Information exchange between anesthesia providers and parturients about neuraxial analgesia risks often occurs in the presence of labor pain. This study examined whether the presence of pain impacted the level of recall of information provided to parturients regarding risks of neuraxial techniques.Design: Single-center, nonrandomized study.Setting: Labor and delivery suite and postpartum patient rooms in a large academic medical center.Patients: Two hundred six primigravidas admitted to our labor and delivery suites and receiving neuraxial analgesia were included.Interventions: Informed consent for epidural and spinal placement was obtained by an obstetric anesthesia resident as per our standard practice. At the time of consent, parturients' self-reported level of pain was recorded.Measurements: After delivery, patients completed a questionnaire asking which risks they recalled. Also queried were patient self-reported levels of anxiety at the time of consent, patient satisfaction with the informed consent process, overall satisfaction with pain control, as well as their preferred method and timing of information exchange.Main Results: Only 20.9% of the 206 participating parturients recalled all risks and none of the distractors. There was no difference in recall between those with pain and those without pain at the time of consent. Women experiencing any pain at the time of consent were more likely to be very satisfied with the communication of risks compared with women without pain (96.2% vs 85.5%, P=.005). There was no difference in the preferred method (P=.780) or timing (P=.779) of discussion of risks between women in active labor compared with women with a scheduled induction of labor.Conclusions: Although parturients' recall of neuraxial risks did not differ based on the existence of labor pain, those having pain reported greater satisfaction with the informed consent process. [ABSTRACT FROM AUTHOR]

Distinguishing between the two enantiomers of a molecule is a challenging task due to their nearly identical physical properties. Time-consuming chromatography methods are typically required for this task, which greatly limits the throughput of analysis. Here we describe a fluorescence-based method for the rapid and high-throughput analysis of both small-molecule enantiopurity and concentration. Our approach relies on selective molecular recognition of one enantiomer of the target molecule using a DNA aptamer, and the ability of aptamer-based biosensors to transduce the presence of a target molecule into a dose-dependent fluorescence signal. The key novel aspect of our approach is the implementation of enantiomeric DNA biosensors, which are synthesized from D- and L-DNA, but labeled with orthogonal fluorophores. According to the principle of reciprocal chiral substrate specificity, these biosensors will bind to opposite enantiomers of the target with equal affinity and selectivity, enabling simultaneous quantification of both enantiomers of the target. Using the previously reported DNA biosensor for L-tyrosinamide (L-Tym), we demonstrate the ability to rapidly and accurately measure both enantiopurity and concentration for mixtures of L- and D-Tym. We also apply our enantiomeric biosensors to the optimization of reaction conditions for the synthesis of D-Tym and provide mathematical modeling to suggest that DNA biosensors having only modest binding selectivity can also be used for fluorescence-based enantiopurity measurement. This research provides a generalizable method for high-throughput analysis of reaction mixtures, which is anticipated to significantly accelerate reaction optimization for the synthesis of high-value chiral small molecules. [ABSTRACT FROM AUTHOR]

Abstract: Aims: The objective was to re-examine endothelial function, insulin mediated vasodilation and glucose extraction in the forearm of patients with type 2 diabetes (T2DM) and matched control subjects (CON) to investigate whether blood flow impairments result from diabetes per se or from concurrent disease. Methods: 18 subjects (10 with T2DM, 8 CON) had graded brachial artery infusions of endothelial dependent (acetylcholine: 15, 30, 60μg/min), endothelial independent (sodium nitroprusside: 1, 3, 10μg/min) and partially endothelial mediated (adenosine: 50, 150, 500μg/min) vasodilators. The protocol was repeated during a hyperinsulinemic clamp. Forearm blood flow and glucose extraction were measured at each dose of vasodilator (with/without insulin). Measurements were also taken in the control arm, reflecting systemic insulin infusion only. Results: Non-insulin mediated increases in bulk forearm blood flow were similar in T2DM and CON. However, insulin mediated forearm blood flow responses and glucose extraction were lower in T2DM versus CON. Conclusion: The vasodilatory effect of insulin is impaired in T2DM although bulk flow capacity is maintained. Insulin mediated glucose extraction is reduced during concomitant maximal stimulation of forearm blood flow with endothelial-dependent vasodilators, despite maintaining flow. This is consistent with previous work that associates T2DM with impaired insulin mediated capillary recruitment. [Copyright &y& Elsevier]

Abstract: Matrix metalloproteinases (MMPs) are essential for normal collagen turnover, recovery from fibrosis, and vascular permeability. In fibrillar collagens, MMP-1, MMP-8, and MMP-13 cleave a specific glycine–isoleucine or glycine–leucine bond, despite the presence of this sequence in other parts of the protein. This cut site specificity has been hypothesized to arise from a unique, relaxed super-secondary structure in this area due to local hydroxyproline poor character. In this study we examined the mechanism of interaction and cleavage of human type III collagen by fibroblast MMP-1 by using a panel of recombinant human type III collagens (rhCIIIs) containing engineered sequences in the vicinity of the cleavage site. Native and recombinant type III collagens had similar biochemical and structural characteristics, as indicated by transmission electron microscopy, circular dichroism spectropolarimetry, melting temperature and hydroxyproline analysis. A single amino acid change at the I785 cleavage site to proline resulted in partial MMP-1 resistance, but cuts were found in novel sites in the original cleavage region. However, the replacement of five Y-position residues by proline in this region, regardless of I785 variation, conferred complete resistance to MMP-1, MMP-8, MMP-13, trypsin, and elastase. MMP-1 had a decreased specific activity towards and reduced cleavage rate of rhCIII I785P but a K m similar to wild-type. Despite the reductions in protease sensitivity, MMP-1 bound to all of the engineered rhCIIIs with comparable affinity, indicating that MMP-1 binding is not sufficient for cleavage. The relaxed tertiary structure in the MMP cleavage region may permit local collagen unwinding by MMP-1 that enables site-specific proteolysis. [Copyright &y& Elsevier]

Gelatin prepared from animal sources is widely used as a stabilizer in vaccine formulations. The disadvantages associated with their use such as heterogeneity and allerginicity, have led to the development of recombinant human gelatins (rhGs) as a substitute. This study focuses on characterizing the structure and monitoring the physical stability of four molecular weights (∼8.5, 25, 50, and 100 kDa) of rhGs as a function of temperature and pH. The information supplied should be useful in predicting the behavior of rhGs under formulation conditions. A number of spectroscopic techniques were employed in this study. Experimental results indicated that the solution properties of all four rhGs were unpredictable with micro-aggregation observed at various pH values. The 8.5 kDa rhG was found to be in a micro-aggregated state at pH 5 while the 25 kDa rhG was found to be more aggregated at pHs 5, 7, and 8. The properties of these aggregates have been analyzed as a function of temperature. The rhGs were also found to complex with the polyanion heparin through electrostatic and nonelectrostatic interactions. The stability of these complexes has been studied as a function of temperature and pH. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 3363–3378, 2007 [ABSTRACT FROM AUTHOR]

Porcine gelatins have been widely used as stabilizers of macromolecular based pharmaceuticals but the mechanism by which they stabilize has not been precisely established. Their variability and immunogenicity, however, make them less than ideal excipients. In this work, we take advantage of the availability of recombinant human gelatins (rhGs) to explore the mechanism by which they may stabilize proteins. Three model recombinant proteins, human serum albumin (HSA), bovine granulocyte colony stimulating factor (bGCSF), and human fibroblast growth factor-20 (FGF-20) that display a range of isoionic points have been selected for this study. The interaction of these model proteins with four different molecular weight rhGs and porcine gelatin was studied using a variety of biophysical techniques including fluorescence, CD and second derivative UV spectroscopy to monitor tertiary and secondary structure as a function of temperature. The 8.5, 25, and 100 kDa rhGs had the greatest effect on conformational and colloidal stability of HSA. The 8.5, 25, and 50 kDa rhGs also increased the T0 of aggregation of bGCSF and FGF-20. Experiments to probe the mechanism of interaction of model proteins with rhGs suggest that the rhGs might interact with the partially unfolded states of target proteins through a combination of electrostatic and other intermolecular mechanisms to inhibit aggregation. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 3304–3315, 2007 [ABSTRACT FROM AUTHOR]

Abstract: As a part of an ongoing medicinal chemistry effort to identify inhibitors of the Hepatitis C Virus RNA replication, we report here the synthesis and biological evaluation of 9-deaza- and 7,9-dideaza-7-oxa-2′-C-methyladenosine. The parent 2′-C-methyladenosine shows excellent intracellular inhibitory activity but poor pharmacokinetic profile. Replacement of the nucleoside-defining 9-N of 2′-C-methyladenosine with a carbon atom was designed to yield metabolically more stable C-nucleosides. Modifications at position 7 were designed to exploit the importance of the hydrogen bond accepting properties of this heteroatom in modulating the adenosine deaminase (ADA) mediated 6-N deamination. 7-Oxa-7,9-dideaza-2′-C-methyladenosine was found to be a moderately active inhibitor of intracellular HCV RNA replication, whereas 9-deaza- 2′-C-methyladenosine showed only weak activity despite excellent overlap of both of the synthesized target compounds with 2′-C-methyladenosine’s three dimensional structure. Position 7 of the nucleobase proved to be an effective handle for modulating ADA-mediated degradation, with the rate of degradation correlating with the hydrogen-bonding properties at this position. [Copyright &y& Elsevier]

Using MRI, we evaluated the degree of involvement of muscles in the lower extremities of 18 unselected patients with facioscapulohumeral muscular dystrophy (FSHD). Findings were correlated with fragment size of the mutated gene, age, disease duration and muscle power. Most affected muscles were the hamstrings followed by the tibialis anterior and the medial gastrocnemius. The vastus-, gluteal- and peroneal muscles were the most unaffected, and the psoas muscle did not show evidence of involvement in any of the investigated subjects. Asymmetric involvement was evident in 15% of the investigated muscles on MRI and 6% on manual muscle strength testing. MRI findings in muscle tended to correlate with disease duration (r = 0.49; p < 0.05), but not with gene fragment size or age. MRI disclosed involvement of muscles performing hip flexion and ankle dorsal flexion that could not be detected by manual muscle strength testing. Otherwise, there was a close correlation (≈ r = 0.75; p < 0.0001) between muscle strength and MRI severity score for other muscle groups. The present study shows that MRI may disclose muscle involvement in FSHD that is not apparent on manual muscle testing, and suggests that MRI of muscle may be an important assessment tool in clinical trials involving patients with FSHD. [ABSTRACT FROM AUTHOR]

The use of genetically engineered microorganisms is a cost-effective, scalable technology for the production of recombinant human collagen (rhC) and recombinant gelatin (rG). This review will discuss the use of yeast ( Pichia pastoris, Saccharomyces cerevisiae, Hansenula polymorpha) and of bacteria ( Escherichia coli, Bacillus brevis) genetically engineered for the production of rhC and rG. P. pastoris is the preferred production system for rhC and rG. Recombinant strains of P. pastoris accumulate properly hydroxylated triple helical rhC intracellularly at levels up to 1.5 g/l. Coexpression of recombinant collagen with recombinant prolyl hydroxylase results in the synthesis of hydroxylated collagen with thermal stability similar to native collagens. The purified hydroxylated rhC forms fibrils that are structurally similar to fibrils assembled from native collagen. These qualities make rhC attractive for use in many medical applications. P. pastoris can also be engineered to secrete high levels (3 to 14 g/l ) of collagen fragments with defined length, composition, and physiochemical properties that serve as substitutes for animal-derived gelatins. The replacement of animal-derived collagen and gelatin with rhC and rG will result in products with improved safety, traceability, reproducibility, and quality. In addition, the rhC and rG can be engineered to improve the performance of products containing these biomaterials. [ABSTRACT FROM AUTHOR]

Insulin-mediated glucose clearance (GC) is diminished in type 2 diabetes. Skeletal muscle has been estimated to account for essentially all of the impairment. Such estimations were based on leg muscle and extrapolated to whole body muscle mass. However, skeletal muscle is not a uniform tissue and insulin resistance may not be evenly distributed. We measured basal and insulin-mediated (1 pmol min−1 kg−1) GC simultaneously in the arm and leg in type 2 diabetes patients (TYPE 2) and controls (CON) (n= 6 for both). During the clamp arterio-venous glucose extraction was higher in CONversusTYPE 2 in the arm (6.9± 1.0versus4.7± 0.8%; mean±s.e.m.;P= 0.029), but not in the leg (4.2± 0.8versus3.1± 0.6%). Blood flow was not different between CON and TYPE 2 but was higher (P<0.05) in armversusleg (CON: 74± 8versus56± 5; TYPE 2: 87± 9versus43± 6 ml min−1 kg−1 muscle, respectively). At basal, CON had 84% higher arm GC (P= 0.012) and 87% higher leg GC (P= 0.016) compared with TYPE 2. During clamp, the difference between CON and TYPE 2 in arm GC was diminished to 54% but maintained at 80% in the leg. In conclusion, this study shows that glucose clearance is higher in arm than leg muscles, regardless of insulin resistance, which may indicate better preserved insulin sensitivity in arm than leg muscle in type 2 diabetes. [ABSTRACT FROM AUTHOR]

Abstract: Gelatin is used as a stabilizer in several vaccines. Allergic reactions to gelatins have been reported, including anaphylaxis. These gelatins are derived from animal tissues and thus represent a potential source of contaminants that cause transmissible spongiform encephalopathies. We have developed a low molecular weight human sequence gelatin that can substitute for the animal sourced materials. A cDNA fragment encoding 101 amino acids of the human proα1 (I) chain was amplified, cloned into plasmid pPICZα, integrated into Pichia pastoris strain X-33, and isolates expressing high levels of recombinant gelatin FG-5001 were identified. Purified FG-5001 was able to stabilize a live attenuated viral vaccine as effectively as porcine gelatin. This prototype recombinant gelatin was homogeneous with respect to molecular weight but consisted of several charge isoforms. These isoforms were separated by cation exchange chromatography and found to result from a combination of truncation of the C-terminal arginine and post-translational phosphorylation. Site-directed mutagenesis was used to identify the primary site of phosphorylation as serine residue 546; serine 543 was phosphorylated at a low level. A new construct was designed encoding an engineered gelatin, FG-5009, with point mutations that eliminated the charge heterogeneity. FG-5009 was not recognized by antigelatin IgE antibodies from children with confirmed gelatin allergies, establishing the low allergenic potential of this gelatin. The homogeneity of FG-5009, the ability to produce large quantities in a reproducible manner, and its low allergenic potential make this a superior substitute for the animal gelatin hydrolysates currently used to stabilize many pharmaceuticals. [Copyright &y& Elsevier]

The tools of recombinant protein expression are now being used to provide recombinant sources of both collagen and gelatin. The primary focus of this review is to discuss alternatives to bovine collagen for biomedical applications. Several recombinant systems have been developed for production of human sequence collagens. Mammalian and insect cells were initially used, but were thought to be too costly for commercial production. Yeast have been engineered to express high levels of type I homotrimer and heterotrimer and type II and type III collagen. Co-expression of collagen genes and cDNAs encoding the subunits of prolyl hydroxylase has lead to the synthesis of completely hydroxylated, thermostable collagens. Human types I and III collagen homotrimers have been expressed in transgenic tobacco plants, while transgenic mice have been engineered to produce full-length type I procollagen homotrimer as well as a α2 (I) homotrimeric mini-collagen. Most recently, a transgenic silkworm system was used to produce a fusion protein containing a collagenous sequence. Each of these transgenic systems holds great promise for the cost-effective large-scale production of recombinant human collagens. As seen in other recombinant expression systems, transgenic silkworms, tobacco, and mice lack sufficient endogenous prolyl hydroxylase activity to produce fully hydroxylated collagen. In mice and tobacco, this was overcome by over-expression of prolyl hydroxylase, analogous to what has been done in yeast and insect cell culture. In addition to recombinant alternatives to bovine collagen, other sources such as fish and sponge collagen are discussed briefly. Recombinant gelatin has been expressed in Pichia pastoris and Hansenula polymorpha in both non-hydroxylated and hydroxylated forms. Pichia was shown to be a highly productive system for gelatin production. The recombinant gelatins produced in yeast are of defined molecular weight and physio-chemical properties and represent a new biomaterial not previously available from animal sources. Genetic engineering has made great progress in the areas of recombinant collagen and gelatin expression, and there are now several alternatives to bovine material that offer an enhanced safety profile, greater reproducibility and quality, and the ability of these materials to be tailored to enhance product performance. [Copyright &y& Elsevier]

It is well known that morphological changes at the cellular level occur in muscle of patients with mitochondrial myopathy (MM), but changes in muscle structure with fat infiltration and gross variation of muscle fiber size with giant fibers, normally encountered in the muscular dystrophies, have typically not been associated with mitochondrial disease. We investigated gross and microscopic muscle morphology in thigh muscles by muscle biopsy and MRI in 16 patients with MM, and compared findings with those obtained in muscular dystrophy patients and healthy subjects. Changes of muscle architecture, similar to those found in the group of muscular dystrophy patients occurred consistently in patients with a high mutation load for single, largescale deletions of mtDNA, but were absent in all patients with the 3243A→G mtDNA point mutation. Dystrophic changes of muscle architecture were also present in one MM patient with a unique, sporadic mutation in the mtDNA tRNAMet gene. These findings provide evidence that morphological changes in muscle of MM patients are common and may resemble those of muscular dystrophies, but that development of dystrophic-like changes in muscle relate to genotype. [ABSTRACT FROM AUTHOR]

Critical examinations of the television program 'The X-Files' support the contention that its formula for success included narrative originality, social commentary and cultural resonance to a degree rare in prime time network commercial entertainment television, but not so uncommon within the tradition of TV science fiction. Science fiction in broadcast history, although neglected critically for its associations with the populist leanings of genre criticism on one hand and the lowbrow reputation of broadcasting on the other, is rich with examples of memorable and enduring programming. It would thus appear not inappropriate to seek to situate 'The X-Files' as the textual benchmark for science fiction TV and film, in relation to the place of science fiction in post September 11, 2001 America.

"Culture wars," a term popularized in the 1980s, has become a part of the grammatical lexicon of both politicos and cultural critics who sense that the U.S. is becoming increasingly polarized. On a general level, this paper studies the polarization enacted by the "external, definable, and transcendent authority" side in the "culture wars" — the New Religious Right (NRR) — as it calls for a "war" against the media. More specifically, this paper surveys one NRR group, Donald Wildmon's American Family Association (AKA), as it fights a battle against MCA/Universal's 'The Last Temptation of Christ,' for the film's "demeaning and degrading portrayal of the life of Christ.

Presents an integrative approach to understanding clergy burnout. Studies on what triggers burnout among clergy; Integration of the Self-psychology of Heinz Kohut with the general systems theory of M. Bowen (1978) to better understand burnout; Implications of the integrative model for the treatment and prevention of burnout.

Modulation of ornithine decarboxylase (ODC) gene expression by retinoids was analyzed in human keratinocyte cultures maintained in serum-free medium containing 0.15 mM Ca++. Cells were incubated with all-trans-retinoic acid, 13-cis-retinoic acid or arotinoid Ro15-0778 (10-10 to 10-5 M), total RNA was isolated, and mRNA transcripts for ODC were analyzed by Northern and slot blot hybridizations with a human ODC cDNA. Treatment of cells for 24 h resulted in a dose-dependent decrease in ODC mRNA levels, with an estimated IC50 of ∼1 × 10-8 M for all-trans- and 13-cis-retinoic acid, while Ro15-0778 was somewhat less effective (IC50 ∼1-5 × 10-7 M). The suppression of ODC mRNA levels by retinoids was detectable at ∼3 h of incubation, with essentially a maximal inhibition at 12 h. Reduced ODC mRNA levels noted after 24 h of incubation with 5 × 10-7 M all-trans-retinoic acid were accompanied by a reduction in ODC enzyme activity. To determine if all-trans-retinoic acid was regulating ODC gene expression directly, or if protein synthesis was required, ODC expression was analyzed in cultures treated with protein synthesis inhibitors. In the presence of cycloheximide or puromycin, all-trans-retinoic acid did not suppress ODC mRNA levels. These findings suggest that suppression of ODC gene expression is not a direct effect of all-trans-retinoic acid, but depends on ongoing protein synthesis. [ABSTRACT FROM AUTHOR]

Basement membrane zone gene expression by fibroblast cultures, established from individuals varying in age from 14 fetal weeks to 61 years, was examined by molecular hybridizations with human sequence specific cDNAs corresponding to type IV procollagen and laminin subunit polypeptides. Northern transfer analysis with poly(A)+ RNA revealed the presence of specific mRNA transcripts for α1 (IV) and α2(IV) chains of type IV procollagen as well as B1 and B2 chains of laminin. Laminin A chain mRNAs were not detected using the same RNA preparations. Quantitative estimates of the steady-state levels of type IV procollagen and laminin mRNAs indicated that they were of relatively low abundance, as compared with mRNA to type I procollagen. The expression of α1(IV) and α2(IV) collagen genes was high in fetal fibroblasts but was reduced to low, yet detectable, levels in cultures established from 3-d to 61-year-old individuals. In contrast, the laminin B1 and B2 chain mRNA levels showed little age-associated variation within the cultures examined. These results provide evidence for differential regulation of the expression of different basement membrane zone macro-molecules during chronologic aging. [ABSTRACT FROM AUTHOR]

The extracellular connective tissue matrix of the skin is a complex aggregate of distinct collagenous and non-collagenous components. Optimal quantities and delicate interactions of these components are necessary to maintain normal physiologic properties of skin. This overview summarizes the progress made in understanding the normal biology and biochemistry of the extracellular matrix, and will highlight cutaneous diseases with underlying molecular defects in the structure and expression of extracellular matrix components. [ABSTRACT FROM AUTHOR]

A human cDNA library was constructed utilizing RNA isolated from cultured skin fibroblasts. Recombinant clones containing elastin sequences were identified by plaque hybridizations with previously characterized human placental elastin cDNAs. Seven positive recombinant clones with inserts of ∼ 3.2-2.2 kb were isolated. Characterization of the clones by restriction endonuclease analysis and dot-blot hybridizations with exon-specific synthetic oligonucleotides demonstrated considerable variability in the primary nucleotide sequence. Dideoxy nucleotide sequencing confirmed this finding. The variability is most likely a result of alternative splicing of exons from the primary elastin transcripts. The two largest clones contained ∼ 1 kb of 3' untranslated sequence and ∼ 2.2 kb of translated sequence encoding 730 amino acids. Six amino acids, encoded by exon 12A, have not been previously noted in human elastin cDNAs. In addition, these human skin fibroblast clones contained a 49 bp 5' Untranslated sequence. These results demonstrate that there is considerable variability in the processed nucleotide sequence of the elastin mRNAs. These transcripts may code for isoforms of tropoelastin with different biologic properties. [ABSTRACT FROM AUTHOR]

Basement membrane zone gene expression by fibroblast cultures established from adult human skin was examined by molecular hybridizations with human sequence-specific cDNAs corresponding to pro-α1(IV) chain of type IV collagen and B2 chain of laminin. Northern transfer analysis of po1y(A)+RNA isolated from fibroblast cultures clearly revealed the presence of specific mRNA transcripts, indicating the expression of the genes coding for pro-α1(IV) and laminin B2 polypeptides. Quantitative estimates of the relative levels of these mRNAs coding for basement membrane zone components indicated that they are of relatively low abundance in control fibroblasts, as compared with mRNAs coding for fibronectin or pro-α1(I) chain of type I procollagen. In fibroblast cultures established from the lesional skin of a 32-year-old patient with lipoid proteinosis, the levels of mRNA coding for pro-α1 (IV) polypeptides were increased approximately 4.5-fold, as compared with age- and passage-matched control cultures. This increase was selective in that the levels of fibronectin, pro-α1 (I), and β-actin mRNAs were unaltered in the same cultures. The increase in pro-α1(IV) mRNA level was also uncoordinate with the expression of the laminin B2 chain gene, which was unaltered in lipoid proteinosis. The selective increase in pro-α1(IV) mRNA may have relevance to the accumulation of this basement membrane component in the skin in lipoid proteinosis. [ABSTRACT FROM AUTHOR]

The article discusses trends of family life in the United States. Teenagers who marry are especially at risk for divorces, but younger couples alone do not account for the high divorce statistics. While the majority of younger men and women do remarry, men tend to marry more often and quicker than women. About half of mothers-only families live at or below the poverty level, compared with only 10 percent of married couples with children. Divorce and single parenting are certainly stressors, which have multidirectional impact on families, but intact marriages are far from carefree. Another troubling trend with regard to families in the United States is the high rate of teenage pregnancy. The United States has the highest rates of teenage pregnancy of any industrialized country, with a disproportionate impact on black teenagers. Black families are more likely to be headed by single females than are white families. Women in the labor force are concentrated into a narrow band of "pink collar" or clerical jobs, and paid significantly less for their contribution to the welfare of the organization.

Background and Objective: During the COVID-19 vaccination campaign, over 34,000 reports of heavy menstrual bleeding following the administration of COVID-19 vaccines originating in the Economic European Area were submitted to EudraVigilance, the European Union database of suspected adverse drug reactions. More than 90% of these reports were sent by consumers while the remaining by healthcare professionals. Public concerns regarding menstruation disorders in COVID-19 vaccinees were also covered by the media. We investigated the impact of media attention on the reporting trends of heavy menstrual bleeding to EudraVigilance. Methods: We used media outlets published in the Economic European Area on menstrual disorders and COVID-19 vaccines from the beginning of the vaccination campaign in the Economic European Area (1 January, 2021) until December 2022 (i.e., after the regulatory request to add the adverse event to the product information) and spontaneous reports from EudraVigilance. Results: We found that the publication of safety updates from regulatory authorities and subsequent coverage in media outlets preceded increased reporting to EudraVigilance. Furthermore, the heavy menstrual bleeding reported in the cases occurred several weeks or months earlier and were not submitted to the respective date. The analysis suggests that the spikes in reporting of heavy menstrual bleeding were to some extent influenced by media coverage in some countries. Conclusions: Consumer reporting to the European Union spontaneous data collection system, EudraVigilance, was of high value for regulatory safety reviews, albeit the reporting behaviours were not free of the influence of the media. These sources of information can be investigated to understand the context of safety concerns of public health interest. [ABSTRACT FROM AUTHOR]

Objective: To compare the effect of a conventional to an intensive blood pressure monitoring regimen on blood pressure in hypertensive patients in the general practice setting.Design: Randomized controlled parallel group trial with 12-month follow-up.Setting: One hundred and ten general practices in all regions of Denmark.Participants: One thousand forty-eight patients with essential hypertension.Intervention: Conventional blood pressure monitoring ('usual group') continued usual ad hoc blood pressure monitoring by office blood pressure measurements, while intensive blood pressure monitoring ('intensive group') supplemented this with frequent home blood pressure monitoring and 24-hour ambulatory blood pressure monitoring.Primary outcome measures: Mean day- and night-time systolic and diastolic 24-hour ambulatory blood pressure.Secondary outcome measures: Change in systolic and diastolic office blood pressure and change in cardiovascular risk profile.Results: Of the patients, 515 (49%) were allocated to the usual group, and 533 (51%) to the intensive group. The reductions in day- and night-time 24-hour ambulatory blood pressure were similar (usual group: 4.6 ± 13.5/2.8 ± 82 mmHg; intensive group: 5.6 ± 13.0/3.5 ± 8.2 mmHg; P = 0.27/P = 0.20). Cardiovascular risk scores were reduced in both groups at follow-up, but more so in the intensive than in the usual group (P = 0.02).Conclusion: An intensive blood pressure monitoring strategy led to a similar blood pressure reduction to conventional monitoring. However, the intensive strategy appeared to improve patients' cardiovascular risk profile through other effects than a reduction of blood pressure.Trial registration: Clinical Trials NCT00244660. [ABSTRACT FROM AUTHOR]

Introduction: Concerns over serious respiratory depression in children led to two European Union (EU) referral procedures (in 2013 and 2015) to review the benefit-risk balance of codeine in this population when used for pain relief, cough or cold. Consequently, codeine should no longer be used in children aged < 12 years and restrictions were introduced for treatment in children ≥ 12 years.Objective: This multinational collaborative study aimed to assess the effectiveness of these risk minimisation measures by evaluating changes in prescribing of codeine and alternative treatments.Method: Children under 12 and 12-18 years old were followed between 2010 and 2017 to analyse quarterly trends in prescribing of codeine and alternative treatments in electronic health records from France, Germany, Norway, Spain and the United Kingdom using interrupted time series analysis.Results: Overall prescribing of codeine in children decreased in all five countries, reaching near zero prevalence in children under 12 years of age. This was accompanied by an increase in use of other opioid analgesics in France (from 0.15 to 0.56 prevalence per 100 person-years immediately after the first referral), Norway (from 0.0006 to 0.0013 at the end of the study), the United Kingdom (from 0.018 to 0.05 at the end of the study), and an increase in non-opioid analgesics in Norway (from 0.045 to 0.075 at the end of the study) after the referral on pain relief indication. The referral on cough/cold indication led to a decrease in use of opioid and non-opioid antitussives in children aged < 12 years in France (from 10 to 7 and 20 to 16, respectively) and had no impact in other countries. Overall prescribing trends for codeine and alternatives were similar across both age groups within each country.Conclusion: The decrease in use of codeine shows that healthcare professionals followed the adopted measures and switched prescribing practices for pain management in children aged < 18 years towards opioid or non-opioid analgesics depending on national clinical and reimbursement settings. Whist the magnitude of the first referral on pain differed between countries, the second referral on cough/cold had only a minimal impact on the use of codeine and antitussives. [ABSTRACT FROM AUTHOR]

Comment cards are a common way to obtain timely customer feedback. Prior research has shown, however, that comment cards are often poorly designed. As such, researchers have identified areas where improvements can be made. Despite these efforts, there are at least three issues where additional clarification is warranted; namely, the appropriate types of questions to ask, question wording related to the use of double-barreled questions, and the appropriate amount of writing space for comments. This research note helps clarify these issues by proposing: (a) that the most appropriate questions are those that help quantify the qualitative comments, (b) that not all double-barreled questions are as problematic as some suggest, and (c) that the minimally acceptable amount of space for writing comments is more than previously thought. The proposed clarifications are illustrated using a sample of restaurant comment cards. [ABSTRACT FROM AUTHOR]

Describe dosing of local anesthetic when both a periarticular injection (PAI) and peripheral nerve block (PNB) are utilized for knee arthroplasty analgesia, and compare the dosing of local to suggested maximum dosing, and look for evidence of local anesthetic systemic toxicity (LAST). A single center retrospective cohort study between May 2018 and November 2022. A major academic hospital. Patients who had both a PAI and PNB while undergoing primary, revision, total, partial, unilateral, or bilateral knee arthroplasty. None. Calculate the dose of local anesthetic given via PAI, PNB, and both routes combined as based on lean body weight and compare that to the suggested maximum dosing. Look for medications, clinical interventions, and critical event notes suggestive of a LAST event. There were 4527 knee arthroplasties where both a PAI and PNB were performed during the study period. When combining PAI and PNB doses, >75% of patients received more than the suggested maximum dose of 3 mg/kg lean body weight. The median local anesthetic dosing over the study period, 4.4 mg/kg (IQR 3.5,5.9), was 147% of the suggested maximum dose (IQR 117,197). There was no conclusive evidence of LAST among any of the patients in the study. Over the course of our study, we had 4527 knee arthroplasties with a median PAI and PNB local anesthetic dose that was 147% of the suggested maximum without any clear clinical evidence of a LAST event. [Display omitted] • Total combined dose of local anesthetic given via PAI and PNB often exceeds suggested maximum lean body weight-based amount. • No definitive local anesthetic toxicity events were observed in this retrospective single-center study. • Maximum recommended doses of bupivacaine and ropivacaine could be reevaluated in knee arthroplasty. [ABSTRACT FROM AUTHOR]

Abstract: The synthesis of 2′-β-C-methyl-neplanocin derivatives is described. The key intermediate cyclopentenyl alcohol 12 is prepared from sugar 5 in 12 steps. Coupling of 12 with appropriately protected purine, 7-deaza pyrimidine, uracil and pyrimidine bases via the Mitsunobu reaction followed by deprotection afforded the target cyclopentenyl nucleosides (18–23, 27). The synthesized compounds were evaluated as potential inhibitors of the hepatitis C virus (HCV) in vitro. Unfortunately, none of them show anti-HCV activity below EC50 100μM. [Copyright &y& Elsevier]

The term “endothelial dysfunction” refers to the inability or attenuated effect of the endothelial cells in participating in the relaxation of the adjacent smooth muscle, thus causing less vasodilation. Although endothelial dysfunction is often seen in patients with type 2 diabetes, it does not necessarily follow that insulin resistance and (or) hyperglycemia is causing the inability to respond properly to vasodilatory stimuli. Rather, this could be related to the impact of concomitant cardiovascular risk factors that are almost invariably present in patients with type 2 diabetes. The impact of physical training — or the opposite, inactivity — on endothelial function is not fully elucidated. Some studies have shown positive effects of physical training, whereas others have not. In general, physical training can improve endothelial function when this is impaired. However, physical training does not seem to have any effect on endothelial function when this is normal. [ABSTRACT FROM AUTHOR]

In the present study we investigated the relationship between plasma fatty acids (FA) and intramuscular triacylglycerol (IMTAG) kinetics of healthy volunteers. With this aim[U-13C]-palmitate was infused for 10 h and FA kinetics determined across the leg. In addition, the rate of FA incorporation into IMTAG in vastus lateralis muscle was determined during two consecutive 4-h periods (2–6 h and 6–10 h). Fifty to sixty per cent of the FA taken up from the circulation were esterified into IMTAG, whereas 32 and 42% were oxidized between 2–6 and 6–10 h, respectively. IMTAG fractional synthesis rate was 3.4± 0.8% h−1 and did not change between the two 4- h periods, despite an increase in arterial FA concentration. IMTAG concentration was also unchanged, implying that the IMTAG fractional synthesis rate was balanced by an equal rate of breakdown. FA oxidation increased over time, which could be due to the observed decline in plasma insulin concentration. In conclusion, a substantial fraction of the fatty acids entering skeletal muscle in post-absorptive healthy individuals is esterified into IMTAG, due to its high turnover rate (29 h pool−1). An increase in FA level, as a consequence of short-term fasting, does not seem to increase IMTAG synthesis rate and pool size. [ABSTRACT FROM AUTHOR]

This study investigates relationship dimensions of 70 Australian couples and tests the specific hypothesis that the difficulties encountered in the stepparenting dimension are more pronounced in complex stepfamilies (with children from both parents) than in simple step families. The findings, based on responses to 125 items comprising 12 ENRICH-A scales, indicate that the major strengths for stepfamilies were the areas of sexual relationship and equalitarian roles, while the major stressors were children and parenting, former attachments, and adjustments. Simple stepfamily couples were in more positive agreement than those from complex stepfamilies across ten other dimensions, including children and parenting, the personality of the partner, communication, and conflict resolution. Recommendations for further research include an exploration of preventive approaches to support stepfamilies. [ABSTRACT FROM PUBLISHER]

Three distinct proteins, namely, β4 integrins, and the 230-kDa (BPAG1) and 180-kDa(BPAG2) bullous pemphigoid antigens, have been shown to co-localize with hemidesmosomes at the dermal-epidermal basement membrane zone. In this study, we examined the expression of the corresponding genes in cultures of normal and transformed human epidermal keratinocytes. The expression of these genes was detected by Northern and in situ hybridizations, and the expression of β4 integrins was also demonstrated by indirect immunofluorescence. The results indicated clearly detectable expression of all three genes in normal keratinocytes, whereas extremely tow or undetectable levels of expression were noted in two transformed cell lines. Addition of TGF- β1 or TGF-β2 (10 ng/ml) up-regulated mRNA levels for all three proteins (up to 4.6 times). The increase by TGF-β1 was particularly striking in keratinocyte cultures incubated in the presence of low (0.15 mM) Ca++, and somewhat less pronounced in the presence of high (1.2 mM) Ca++, The increase in β4 integrin synthesis was also documented by enhanced immunosignal of the corresponding epitopes. These results indicate that the three hemidesmosomal genes studied here are all responsive to TGF-β. These observations, together with precious data on the effects of TGF-β on other components of the skin, suggest that this cytokine may play a role in the development and repair of the cutaneous basement membrane zone. [ABSTRACT FROM AUTHOR]

The long-term survival of biomaterial implants is often hampered by surgery-induced inflammation that can lead to graft failure. Considering that most corneas receiving grafts are either pathological or inflamed before implantation, the risk of rejection is heightened. Here, we show that bioengineered, fully synthetic, and robust corneal implants can be manufactured from a collagen analog (collagen-like peptide-polyethylene glycol hybrid, CLP-PEG) and inflammation-suppressing polymeric 2-methacryloyloxyethyl phosphorylcholine (MPC) when stabilized with the triazine-based crosslinker 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride. The resulting CLP-PEG-MPC implants led to reduced corneal swelling, haze, and neovascularization in comparison to CLP-PEG only implants when grafted into a mini-pig cornea alkali burn model of inflammation over 12 months. Implants incorporating MPC allowed for faster nerve regeneration and recovery of corneal sensation. CLP-PEG-MPC implants appear to be at a more advanced stage of regeneration than the CLP-PEG only implants, as evidenced by the presence of higher amounts of cornea-specific type V collagen, and a corresponding decrease in the presence of extracellular vesicles and exosomes in the corneal stroma, in keeping with the amounts present in healthy, unoperated corneas. Simpson, McTiernan et al. develop fully synthetic corneal implants. The resulting CLP-PEG-MPC implants show reduced corneal swelling, haze, and neovascularization compared to CLP-PEG only implants when grafted into a mini-pig cornea alkali burn model of inflammation over 12 months. This study suggests the superiority of CLP-PEG-MPC implants over the CLP-PEG only implants. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research and antiviral discovery are hampered by the lack of a cellbased virus replication system that can be readily adopted without biosafety level 3 (BSL-3) restrictions. Here, the construction of a noninfectious SARS-CoV-2 reporter replicon and its application in deciphering viral replication mechanisms and evaluating SARS-CoV-2 inhibitors are presented. The replicon genome is replication competent but does not produce progeny virions. Its replication can be inhibited by RdRp mutations or by known SARS-CoV-2 antiviral compounds. Using this system, a high-throughput antiviral assay has also been developed. Significant differences in potencies of several SARS-CoV-2 inhibitors in different cell lines were observed, which highlight the challenges of discovering antivirals capable of inhibiting viral replication in vivo and the importance of testing compounds in multiple cell culture models. The generation of a SARS-CoV-2 replicon provides a powerful platform to expand the global research effort to combat COVID-19. [ABSTRACT FROM AUTHOR]

Since the initial report of the novel Coronavirus Disease 2019 (COVID-19) emanating from Wuhan, China, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has spread globally. While the effects of SARS-CoV-2 infection are not completely understood, there appears to be a wide spectrum of disease ranging from mild symptoms to severe respiratory distress, hospitalization, and mortality. There are no Food and Drug Administration (FDA)-approved treatments for COVID-19 aside from remdesivir; early efforts to identify efficacious therapeutics for COVID-19 have mainly focused on drug repurposing screens to identify compounds with antiviral activity against SARS-CoV-2 in cellular infection systems. These screens have yielded intriguing hits, but the use of nonhuman immortalized cell lines derived from non-pulmonary or gastrointestinal origins poses any number of questions in predicting the physiological and pathological relevance of these potential interventions. While our knowledge of this novel virus continues to evolve, our current understanding of the key molecular and cellular interactions involved in SARS-CoV-2 infection is discussed in order to provide a framework for developing the most appropriate in vitro toolbox to support current and future drug discovery efforts. [ABSTRACT FROM AUTHOR]

Case: A 29-year-old man was admitted with acute atraumatic left hip pain and inability to bear weight. Subsequent workup revealed an intracapsular pathologic fracture of the femoral neck secondary to a high-grade spindle cell sarcoma. A unique method of extra-articular resection and reconstruction using a 3-dimensional (3D)-printed custom cutting jig and a custom acetabular component was pursued. Wide margins were achieved without violating the joint capsule but preserving pelvic integrity, allowing a return to an active lifestyle. Conclusion: Three-dimensional-printed custom resection guides and implants can help achieve adequate resection margins while preserving pelvic integrity and function. [ABSTRACT FROM AUTHOR]

The article discusses about the transplantation with donor corneas as the mainstay for treating corneal blindness, but a severe worldwide shortage necessitates the development of other treatment options. It mentions that LiQD Cornea as an alternative to conventional corneal transplantation and sealants, along with information on the corneal perforation from infection or inflammation as sealed with cyanoacrylate glue.

Background: Intrathecal opioids are routinely administered during spinal anesthesia for postcesarean analgesia. The effectiveness of intrathecal morphine for postcesarean analgesia is well established, and the use of intrathecal hydromorphone is growing. No prospective studies have compared the effectiveness of equipotent doses of intrathecal morphine versus intrathecal hydromorphone as part of a multimodal analgesic regimen for postcesarean analgesia. The authors hypothesized that intrathecal morphine would result in superior analgesia compared with intrathecal hydromorphone 24 h after delivery. Methods: In this single-center, double-blinded, randomized trial, 138 parturients undergoing scheduled cesarean delivery were randomized to receive 150 µg of intrathecal morphine or 75 µg of intrathecal hydromorphone as part of a primary spinal anesthetic and multimodal analgesic regimen; 134 parturients were included in the analysis. The primary outcome was the numerical rating scale score for pain with movement 24 h after delivery. Static and dynamic pain scores, nausea, pruritus, degree of sedation, and patient satisfaction were assessed every 6 h for 36 h postpartum. Total opioid consumption was recorded. Results: There was no significant difference in pain scores with movement at 24 h (intrathecal hydromorphone median [25th, 75th] 4 [3, 5] and intrathecal morphine 3 [2, 4.5]) or at any time point (estimated difference, 0.5; 95% CI, 0 to 1; P = 0.139). Opioid received in the first 24 h did not differ between groups (median [25th, 75th] oral morphine milligram equivalents for intrathecal hydromorphone 30 [7.5, 45.06] vs. intrathecal morphine 22.5 [14.0, 37.5], P = 0.769). From Kaplan–Meier analysis, the median time to first opioid request was 5.4 h for hydromorphone and 12.1 h for morphine (log-rank test P = 0.200). Conclusions: Although the hypothesis was that intrathecal morphine would provide superior analgesia to intrathecal hydromorphone, the results did not confirm this. At the doses studied, both intrathecal morphine and intrathecal hydromorphone provide effective postcesarean analgesia when combined with a multimodal analgesia regimen. In a randomized, double-blinded trial, intrathecal hydromorphone and intrathecal morphine were compared in women receiving cesarean delivery using pain score at 24 h as the primary outcome. The analgesia provided by morphine was not superior to that provided by hydromorphone. In addition, breakthrough analgesic requirements were similar for the two groups. [ABSTRACT FROM AUTHOR]

Identification of new druggable protein targets remains the key challenge in the current antimalarial development efforts. Here we used mass-spectrometry-based cellular thermal shift assay (MS-CETSA) to identify potential targets of several antimalarials and drug candidates. We found that falcilysin (FLN) is a common binding partner for several drug candidates such as MK-4815, MMV000848, and MMV665806 but also interacts with quinoline drugs such as chloroquine and mefloquine. Enzymatic assays showed that these compounds can inhibit FLN proteolytic activity. Their interaction with FLN was explored systematically by isothermal titration calorimetry and X-ray crystallography, revealing a shared hydrophobic pocket in the catalytic chamber of the enzyme. Characterization of transgenic cell lines with lowered FLN expression demonstrated statistically significant increases in susceptibility toward MK-4815, MMV000848, and several quinolines. Importantly, the hydrophobic pocket of FLN appears amenable to inhibition and the structures reported here can guide the development of novel drugs against malaria. [Display omitted] • MS-CETSA identifies falcilysin (FLN) as a common target for several antimalarials • Biochemical assays validate FLN interaction to antimalarials • Crystallography uncovered a hydrophobic pocket that appears amenable to inhibition • FLN conditional knockdown can modulate drug susceptibility Wirjanata et al. use MS-CETSA for screening of antimalarials and found falcilysin, an essential P. falciparum metalloprotease, to be a common target. Biochemical assays successfully validated falcilysin-drug binding and enzymatic activity inhibition, whereas X-ray crystallography further uncovered a shared hydrophobic pocket in falcilysin that appears amenable to inhibition. [ABSTRACT FROM AUTHOR]

Objective: Sporadic fatal adverse events have been reported during treatment of multiple sclerosis with alemtuzumab. To provide a systematic overview, we searched the centralized European Medicines Agency database of suspected adverse reactions related to medicinal products (EudraVigilance) for fatal adverse events associated with treatment with alemtuzumab (Lemtrada®) for multiple sclerosis. Four independent reviewers with expertise on MS, clinical immunology, infectious diseases and clinical pharmacology reviewed the reports, and scored the likelihood for causality. Results: We identified nine cases with a probable and one case with a possible causal relationship between alemtuzumab treatment and a fatal adverse event. Six of these patients died within one month after treatment; one from intracerebral hemorrhage, two from acute multiple organ failure and septic shock, one from listeriosis, one from pneumonia and one from agranulocytosis. Four patients died several months after administration of alemtuzumab from either autoimmune hepatitis, immune-mediated thrombocytopenia, autoimmune hemolytic anemia or agranulocytosis. Four of the 10 cases had been published previously in case reports or congress abstracts. Fatal adverse events related to treatment with alemtuzumab occur more frequently than previously published in the literature. A significant proportion occurs in the first month after treatment. [ABSTRACT FROM AUTHOR]

Abstract: Ciguatera fish poisoning is a seafood‐toxin illness resulting from consumption of fish contaminated with ciguatoxins. Managing ciguatera fish poisoning is complex. It is made easier, however, by local fishers from endemic areas reporting regional predictability for local fish species’ ciguatera fish poisoning risk, which the present study then tested. We investigated the prevalence of ciguatoxins in 4 commonly marketed and consumed species (Balistes vetula, Haemulon plumierii, Ocyurus chrysurus, and Epinephelus guttatus) across an oceanic gradient (north, south, east, and west) from the US Virgin Islands. Fish muscle extracts were analyzed for Caribbean ciguatoxins using an in vitro mouse neuroblastoma (N2a) cytotoxicity assay and confirmed by liquid chromatography–tandem mass spectrometry (LC‐MS/MS). Fish collected from the north location had 0 fish with detectable ciguatoxins; this site also had the greatest wave energy. Caribbean ciguatoxins in fish ranged from 0.01 to 0.11, 0.004 to 0.10, and 0.005 to 0.18 ng Caribbean ciguatoxin‐1 eq/g, from the west, east, and south respectively. Ciguatoxin‐like activity was detectable by the N2a assay in 40, 41, 50, and 70% of H. plumierii, O. chrysurus, B. vetula, and E. guttatus, respectively. Of the fish collected, 4% had Caribbean ciguatoxin levels exceeding the US Food and Drug Administration guidance of 0.1 ng Caribbean ciguatoxin‐1 eq/g fish. These findings concurred with spatial ciguatera fish poisoning prevalence information provided by local fishers in the US Virgin Islands and demonstrate how partnerships between researchers and fishers can aid the improvement of science‐based ciguatera fish poisoning management. Environ Toxicol Chem 2018;39:1852–1863. Published 2018 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America. [ABSTRACT FROM AUTHOR]

Review: [51 refs. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate and model the pharmacokinetic and pharmacodynamic behavior in rats of FG-3019, a human monoclonal antibody targeting connective tissue growth factor (CTGF). Methods: FG-3019, human CTGF (rhCTGF), or the N-terminal domain of rhCTGF were administered intravenously to rats and concentrations of these proteins as well as endogenous CTGF were determined by immunoassays. FG-3019, or I-labeled FG-3019, and human CTGF (rhCTGF) were co-administered to assess the impact of CTGF on the elimination rate and tissue localization of FG-3019, which was further characterized by immunohistochemical analysis. A PK/PD model for target-mediated elimination of FG-3019 was developed to fit the kinetic data. Results: FG-3019 exhibited non-linear pharmacokinetics in rats. Circulating concentrations of the N-terminal half of CTGF increased after dosing with FG-3019, reached maximal levels after 1-5 days, and returned toward baseline levels as FG-3019 cleared from the circulation, whereas the concentration of intact CTGF was unaffected by administration of FG-3019. Co-administration of rhCTGF dramatically enhanced the rate of FG-3019 elimination, redistributing the majority of I-labeled FG-3019 from the blood to the liver, kidney, spleen and adrenal gland. FG-3019 co-administered with CTGF was found along the sinusoids of the liver and adrenal glands, the capillaries of the kidney glomeruli and in the spleen. A pharmacokinetic model for target-mediated elimination of FG-3019 was used to fit the time courses of FG-3019 and endogenous CTGF plasma concentrations, as well as time courses of rhCTGF and rhCTGF N-fragment after intravenous administration of these species. Conclusions: FG-3019 is subject to target mediated elimination in rats. [ABSTRACT FROM AUTHOR]

Summary Resistance to existing classes of antibiotics drives the need for discovery of novel compounds with unique mechanisms of action. Nargenicin A1, a natural product with limited antibacterial spectrum, was rediscovered in a whole-cell antisense assay. Macromolecular labeling in both Staphylococcus aureus and an Escherichia coli tolC efflux mutant revealed selective inhibition of DNA replication not due to gyrase or topoisomerase IV inhibition. S. aureus nargenicin-resistant mutants were selected at a frequency of ∼1 × 10 −9 , and whole-genome resequencing found a single base-pair change in the dnaE gene, a homolog of the E. coli holoenzyme α subunit. A DnaE single-enzyme assay was exquisitely sensitive to inhibition by nargenicin, and other in vitro characterization studies corroborated DnaE as the target. Medicinal chemistry efforts may expand the spectrum of this novel mechanism antibiotic. [ABSTRACT FROM AUTHOR]