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The human vascular system, comprising endothelial cells (ECs) and mural cells, covers a vast surface area in the body, providing a critical interface between blood and tissue environments. Functional differences exist across specific vascular beds, but their molecular determinants across tissues remain largely unknown. In this study, we integrated single-cell transcriptomics data from 19 human organs and tissues and defined 42 vascular cell states from approximately 67,000 cells (62 donors), including angiotypic transitional signatures along the arterial endothelial axis from large to small caliber vessels. We also characterized organotypic populations, including splenic littoral and blood–brain barrier ECs, thus clarifying the molecular profiles of these important cell states. Interrogating endothelial–mural cell molecular crosstalk revealed angiotypic and organotypic communication pathways related to Notch, Wnt, retinoic acid, prostaglandin and cell adhesion signaling. Transcription factor network analysis revealed differential regulation of downstream target genes in tissue-specific modules, such as those of FOXF1 across multiple lung vascular subpopulations. Additionally, we make mechanistic inferences of vascular drug targets within different vascular beds. This open-access resource enhances our understanding of angiodiversity and organotypic molecular signatures in human vascular cells, and has therapeutic implications for vascular diseases across tissues.

This paper presents a new challenge under study in Brazil: the offshore storage of natural gas and CO2in salt (halite) caverns opened by solution mining in shallow and ultradeep water. The salt rock mechanics in Brazil started with the underground mining of sylvinite (NaCl.KCL) overlying tachyhydrite (CaCl2.MgCl2.12H2O) and research initiated in the 1970s (Costa 1984), to enable the mining of this ore. Applying a numerical simulation of drilling and excavation, the exploration and operation of mines were possible for more than 35 years. The same technology has also been used in the stability analysis of several salt caverns opened by solution mining of halite (NaCl). With the discovery of the giant pre-salt reservoirs in Brazil, underlying about 2000 m of stratified salt rocks, with the presence of tachyhydrite, all the knowledge acquired in the conventional and solution mining designs was ready to be used. More than 200 pre-salt wells have been designed and drilled successfully using the same methodology and computer codes. These approaches are proposed here as a solution to the presented challenge.

The function of a cell is defined by its intrinsic characteristics and its niche: the tissue microenvironment in which it dwells. Here we combine single-cell and spatial transcriptomics data to discover cellular niches within eight regions of the human heart. We map cells to microanatomical locations and integrate knowledge-based and unsupervised structural annotations. We also profile the cells of the human cardiac conduction system1. The results revealed their distinctive repertoire of ion channels, G-protein-coupled receptors (GPCRs) and regulatory networks, and implicated FOXP2in the pacemaker phenotype. We show that the sinoatrial node is compartmentalized, with a core of pacemaker cells, fibroblasts and glial cells supporting glutamatergic signalling. Using a custom CellPhoneDB.org module, we identify trans-synaptic pacemaker cell interactions with glia. We introduce a druggable target prediction tool, drug2cell, which leverages single-cell profiles and drug–target interactions to provide mechanistic insights into the chronotropic effects of drugs, including GLP-1 analogues. In the epicardium, we show enrichment of both IgG+and IgA+plasma cells forming immune niches that may contribute to infection defence. Overall, we provide new clarity to cardiac electro-anatomy and immunology, and our suite of computational approaches can be applied to other tissues and organs.

Cardiovascular disease is the leading cause of death globally. An advanced understanding of cardiovascular disease mechanisms is required to improve therapeutic strategies and patient risk stratification. State-of-the-art, large-scale, single-cell and single-nucleus transcriptomics facilitate the exploration of the cardiac cellular landscape at an unprecedented level, beyond its descriptive features, and can further our understanding of the mechanisms of disease and guide functional studies. In this Review, we provide an overview of the technical challenges in the experimental design of single-cell and single-nucleus transcriptomics studies, as well as a discussion of the type of inferences that can be made from the data derived from these studies. Furthermore, we describe novel findings derived from transcriptomics studies for each major cardiac cell type in both health and disease, and from development to adulthood. This Review also provides a guide to interpreting the exhaustive list of newly identified cardiac cell types and states, and highlights the consensus and discordances in annotation, indicating an urgent need for standardization. We describe advanced applications such as integration of single-cell data with spatial transcriptomics to map genes and cells on tissue and define cellular microenvironments that regulate homeostasis and disease progression. Finally, we discuss current and future translational and clinical implications of novel transcriptomics approaches, and provide an outlook of how these technologies will change the way we diagnose and treat heart disease.

Objective: To investigate perspectives of diabetes, nutrition, and health among underrepresented Mexican-origin males and elucidate on contributing factors of type 2 diabetes disparities. Research Design: Audio-recorded, semi-structured interviews were conducted with 15 native Spanish-speaking, Mexican-origin males. Men were between the ages of 40 and 64 and at risk for type 2 diabetes—defined as >40 years of age, BMI >25, and sedentary over the last 3 months. Data collected included clinical measures (height, weight, waist circumference, blood pressure) and demographic characteristics (years living in the U.S., marital status, employment, occupational physical activity, income, educational attainment, and average physical activity levels). The audio recordings, each lasting approximately 60 min, were transcribed verbatim and underwent three-cycle coding with analysis for codebook formation, categorization, and thematic extraction. Results: Barriers to engaging in health behaviors among Mexican-origin males include poor understanding of diabetes and nutrition, unreliable sources of health information, and socioeconomic status. Enablers of health neglect include intense perceptions of diabetes severity and a reactive health culture rooted in medical machismo and valemadrismo. Perspectives of personal responsibility and the desire to learn through culturally specific recommendations detail potential facilitators of health behaviors. Conclusion: The data convey a network of sociocultural factors that inhibit health prioritization and promote disease misconceptions among Mexican-origin males. Pragmatic, culturally tailored education represents serviceable strategies for health promotion and diabetes prevention. These findings explicate cultural considerations for educators and clinicians seeking optimization of programs and clinical interactions with Mexican-origin men.

This paper presents a proposal for an experimental salt cavern in offshore ultra-deep water for CO2abatement, including the instrumentation plan and well conceptual design evaluated for carbon capture and storage (CCS) application. These studies are based on applied computational mechanics associated with field experimentation that has contributed to the technical feasibility of the underground potash mine at the State of Sergipe in Brazil. This knowhow allowed the stability analysis of several salt caverns for brine production at the State of Alagoas in Brazil and to the drilling through stratified thick layers of salt of the pre-salt reservoirs in Santos Basin. Now, this knowledge has been applied in the design of onshore and offshore salt caverns opened by dissolution for storage of natural gas and CO2. The geomechanical study, through the application of computational mechanics, of offshore giant salt caverns of 450 m high by 150 m in diameter, shows that one cavern can store about 4 billion Sm3or 7.2 million tons of CO2. Before the construction of the giant cavern, which will be the first gas storage offshore in the world, it has been decided to develop an experimental one, with smaller size, to obtained field parameters. The experimental cavern will allow the calibration of parameters to be used in the structural integrity analysis of the cavern and well for storage of natural gas which is rich in CO2under high pressure.

The aim of this paper is to study the potential of using the building information modelling (BIM) technology to assist the activities performed by the team that is responsible for supervising the execution of the federal public building’s contracts in Brazil. The paper describes, in short, the norms and the legislation of the external control body (Court of Auditors), as well as the federal norms and legislation, to understand what should be expected from the organizations’ activities, as well as their obligations. In addition, the most frequent irregularities in public construction were pointed out; the main contributions of BIM, regarding each irregularity, were also identified to avoid the problem. The innovative aspects of this study are the suggestions that were proposed for the use of BIM’s guidelines in public construction supervision in Brazil.

In a world of energy transition and net zero pledges, low-carbon energy solutions are central in the fight against climate change. Hydrogen plays a key role in a clean energy future, but there are still economic and infrastructural challenges to be overcome. Hydrogen storage in large volumes is one of the value chains current limitations. This paper presents a feasibility study of an offshore blue or green hydrogen storage in salt caverns created by leaching, within potentially identifiable salt deposits extent of the Gulf of Mexico coastline (US). In this case study, the hydrogen station will have caverns to store hydrogen and some caverns to dispose of CO2from the blue hydrogen (Carbon Capture and Storage – CCS technology). The construction design includes a fixed platform for the H2HUB and a jack-up platform for drilling. It presents the conceptual design of wells for leaching the caverns and hydrogen injection and withdrawal cycles.

O estudo objetivou identificar a presença da desmotivação na educação física escolar, por meio da teoria da autodeterminação, com vistas à compreensão de possibilidades de intervenção. Aplicamos o questionário de Goudas, Biddle e Fox (1994) para 371 alunos de educação física do ensino médio. Na análise dos dados usamos o teste de Mann‐Whitney e o alphade Conbrach.Os resultados indicaram baixa prevalência de desmotivação (7,3%); evidenciaram que para esse contexto de estudo a motivação prevaleceu e contrariaram achados anteriores. A pesquisa aponta algumas estratégias que podem auxiliar no processo de ensino‐aprendizagem.

In recent years, the antitumoral activity of antimicrobial peptides (AMPs) has been the goal of many research studies. Among AMPs, gomesin (Gm) displays antitumor activity by unknown mechanisms. Herein, we studied the cytotoxicity of Gm in the Chinese hamster ovary (CHO) cell line. Furthermore, we investigated the temporal ordering of organelle changes and the dynamics of Ca2+signaling during Gm-induced cell death. The results indicated that Gm binds to the plasma membrane and rapidly translocates into the cytoplasm. Moreover, 20 μM Gm increases the cytosolic Ca2+and induces membrane permeabilization after 30 min of treatment. Direct Ca2+measurements in CHO cells transfected with the genetically encoded D1-cameleon to the endoplasmic reticulum (ER) revealed that Gm induces ER Ca2+depletion, which in turn resulted in oscillatory mitochondrial Ca2+signal, as measured in cells expressing the genetically encoded probe to the mitochondrial matrix mitPericam. This leads to mitochondria disruption, loss of mitochondrial membrane potential and increased reactive oxygen species prior to membrane permeabilization. Gm-induced membrane permeabilization by a Ca2+-dependent pathway involving Gm translocation into the cell, ER Ca2+depletion and disruption, mitochondrial Ca2+overload and oxidative stress.

A pesar de la eficacia comprobada acerca del uso de warfarina como terapia anticoagulante, las múltiples interacciones, el estrecho margen terapéutico y la necesidad de monitorización seriada han planteado el escenario para el desarrollo de nuevos medicamentos como dabigatran, rivaroxaban y apixaban que ofrecen nuevas alternativas para el tratamiento del paciente anticoagulado. El conocimiento de las características farmacológicas y farmacocinéticas así como el contexto específico en el cual pueden usarse se convierten en una necesidad para los médicos que se enfrentan a este tipo de pacientes.

Passion fruit (Passiflora edulis Sims f. flavicarpa) is popularly known for its sedative and calming properties and is consumed as a fresh fruit or as a juice. The clinical observation of blood incoagulability associated with excessive consumption of passion fruit juice, in a patient treated with warfarin, prompted the current study to investigate in vitro the presence of blood clotting inhibitors in Passiflora edulis Sims f. flavicarpa extract. After purification process, two compounds of distinct molecular weight and inhibitory action were better characterized. One is a trypsin inhibitor similar to inhibitors from Bowman-Birk family, named PeTI-I12, and other is a compound active in coagulation that prolongs aPTT and PT, but does not change TT. The aim of this study is to provide evidence that passion fruit extract’s components play a role on hemostasis and therefore may be relevant in the handling of patients treated with anticoagulants or suffering hemorrhagic diseases.

α‐Tocopherol induces expansion of stem/progenitor cells by non‐identified intracellular pathway and granulocytic/monocytic differentiation through ERK1/2 activation. Tocopherols promote or inhibit growth in different cell types. In the hematopoietic system, the radioprotective property of tocopherols is thought to act through the expansion of primitive hematopoietic cells. However, the mechanisms activated by tocopherols and which HPs are affected remain poorly understood. To better address these questions, mice were treated with α‐tocopherol, and its effects were investigated in the BM microenvironment. α‐Tocopherol induced increased proliferation in HSC/HP cells, leading to BM hyperplasia. In addition, differentiation to the granulocytic/monocytic lineage was enhanced by α‐tocopherol treatment. α‐Tocopherol treatment resulted in decreased basal phosphorylation of ERK1/2, PKC, and STAT‐5 in HSC/HP cells. In contrast, α‐tocopherol enhanced ERK1/2 activation in response to IL‐3 stimulation in HSC/HP cells without altering the expression of IL‐3Rs. Moreover, α‐tocopherol‐induced differentiation and ERK1/2 activation were abolished in mice pretreated with a MEK inhibitor (PD98059); however, pretreatment with PD98059 did not reduce the α‐tocopherol‐mediated increase in HSC/HP cells but instead, further enhanced their proliferation. Therefore, α‐tocopherol induces expansion of HSC/HP cells by a nonidentified intracellular pathway and granulocytic/monocytic differentiation through ERK1/2 activation.

Las manifestaciones en el sistema nervioso central por Listeria monocytogenesson variadas, e incluyen desde la meningitis hasta las presentaciones más infrecuentes como los abscesos cerebrales. Éstos son clínicamente indistinguibles de otros tipos de abscesos y sugieren una alteración de la inmunidad celular. Se describe el caso de una mujer seropositiva para el virus de la inmunodeficiencia humana (VIH) que consultó al servicio de urgencias con un síndrome convulsivo tónico-clónico generalizado de un mes de evolución y deterioro progresivo de su estado de conciencia. En el estudio del líquido cefalorraquídeo se encontró pleocitosis linfocitaria, con elevación de las proteínas e hipoglucorraquia. En el estudio del sistema nervioso central por resonancia magnética se observaron múltiples lesiones hipodensas que se realzaban con el medio contraste. Se aisló L. monocytogenesen el cultivo del líquido cefalorraquídeo. Después de un esquema de tratamiento antibiótico específico asociado a esteroides, los síntomas neurológicos mejoraron y, concomitantemente, se observó una disminución progresiva hasta la desaparición de las lesiones en el seguimiento imaginológico del sistema nervioso central. L. monocytogeneses causa de abscesos cerebrales en pacientes inmunosuprimidos, en especial, con VIH sin terapia antirretroviral o sin profilaxis con trimetoprim-sulfametoxazol.

A recombinant Haematobia irritans irritans trypsin inhibitor (HiTI - Mw 7030 kDa) phagemid library was constructed and displayed functionally on the tip of the filamentous M13 phage. A combinatorial library of 7.2 x 106 mutants was created with HiTI mutations restricted to the P1 - P3 and P5 positions of the reactive site. This combinatorial library was selected for trypsin-like Pr2 proteases of Metarhizium anisopliae fungus, and 11 HiTI mutants containing the following substitutions: K17G, S18R, D19G, S21A, among 60 sequenced clones, were obtained. In order to confirm the inhibitory activity of the selected sequences, we transferred the selected sequence to the shortest protease inhibitor, the sunflower trypsin inhibitor (SFTI - Mw 1533 Da), for inhibitory activity analysis. The hybrid peptide containing the mutated sequence (SFTI-Mut, GRCTRGRGLACFPD-NH2; Ki 14 M) presented an apparent inhibition constant (Kiapp) for Pr2 proteases ~20-fold lower than the control peptide containing the original HiTI sequence (SFTIHiTI, GRCTRKSDLSCFPD-NH2; Ki 259 M). In conclusion, the present work enabled the selection of a specific HiTI mutant for Pr2 proteases of M. anisopliae fungus using a HiTI combinatorial library on M13 phage surface. Selection of strong binders by phage display and their validation as inhibitors using synthetic hybrid peptides proved to be a powerful technique to generate specific serine protease inhibitors suitable for studies of drug design and enzyme-inhibitor interaction.

Abstract: In this work the effect of radical species generated by gamma ray irradiation of aqueous solution upon structure of vasoactive peptide bradykinin (BK, RPPGFSPFR) was investigated. Increasing doses of 1–15 kGy Co60 gamma radiation were applied to BK solutions and a progressive degradation of its structure in a non-linear mode was observed. Two main peptide derivatives generated by these treatments were isolated and characterized through a combined amino acid analysis and daughter ion scanning mass spectrometry approach. Notably, it was observed that only the Phe residue located at position 8 and not 5 of BK was oxidized by reactive hydroxyl radical species given rise to Tyr8-BK and m-Tyr8-BK analogues. Comparative circular dichroism (CD) experiments of these peptides revealed that BK presents greater conformational similarity to Tyr8-BK than to m-Tyr8-BK. These results are in agreement with the biological potencies of these compounds measured in rat uterus and guinea pig ileum muscle contractile experiments. In summary, gamma irradiation of BK solutions revealed a residue- and surprisingly, position-structural modification effect of reactive radicals even in small peptides. Also of value for peptide chemistry field, the approach of applying controlled strong electromagnetic radiation in solution seems to be an alternative and unique strategy for generating, in some cases, peptides derivatives with uncommon structures and valuable for their further therapeutic potential evaluations.

N-Acetyltransferase 2 (NAT2) metabolizes a variety of xenobiotics that includes many drugs, chemicals and carcinogens. This enzyme is genetically variable in human populations and polymorphisms in the NAT2 gene have been associated with drug toxicity and efficacy as well as cancer susceptibility. Here, we have focused on the identification of NAT2 variants in Brazilian individuals from two different regions, Rio de Janeiro and Goiás, by direct sequencing, and on the characterization of new haplotypes after cloning and re-sequencing. Upon analysis of DNA samples from 404 individuals, six new SNPs (c.29T>C, c.152G>T, c.203G>A, c.228C>T, c.458C>T and c.600A>G) and seven new NAT2 alleles were identified with different frequencies in Rio de Janeiro and Goiás. All new SNPs were found as singletons (observed only once in 808 genes) and were confirmed by three independent technical replicates. Molecular modeling and structural analysis suggested that p.Gly51Val variant may have an important effect on substrate recognition by NAT2. We also observed that amino acid change p.Cys68Tyr would affect acetylating activity due to the resulting geometric restrictions and incompatibility of the functional group in the Tyr side chain with the admitted chemical mechanism for catalysis by NATs. Moreover, other variants, such like p.Thr153Ile, p.Thr193Met, p.Pro228Leu and p.Val280Met, may lead to the presence of hydrophobic residues on NAT2 surface involved in protein aggregation and/or targeted degradation. Finally, the new alleles NAT2*6H and NAT2*5N, which showed the highest frequency in the Brazilian populations considered in this study, may code for a slow activity. Functional studies are needed to clarify the mechanisms by which new SNPs interfere with acetylation.

Peptides derived from endogenous hemoglobin play important biological roles in a variety of living systems. In previous works we showed that the fragment 33–61 of bovine α‐hemoglobin (Hb33‐61) and its C‐terminus amidated analogue (Hb33‐61a) exhibit antimicrobial activity and we determined the 3D structure of Hb33‐61a bound to sodium dodecyl sulfate micelles. Here we report that Hb33‐61a is lethal to Candida albicans at 6.25 μM probably through disruption of its plasma membrane. In addition, we show that, even when used at 50 μM, Hb33‐ 61a produces low hemolysis (16% ± 3.0%). Recognizing that one of the key steps to study new compounds with potential pharmaceutical application is to identify the structural elements essential to express biological activity, we also investigated the anticandidal activity of Hb33‐ 61a fragments. The results indicated that Hb40‐61a exhibits the same minimal inhibitory concentration as Hb33‐61a, whereas Hb33‐52a and Hb48‐61a are significantly less active. Noteworthy, for all the peptides tested, we observed that C‐terminus amidation produces a potentiation of their anticandidal activity and we associate that increased biological activity to a preferred structural and spatial organization of the C‐terminal region favored by amidation. Finally, the data show that the most active peptides (Hb33‐61a and Hb40‐61a) are characterized by a central hinge joining the C‐terminal region that presents, containing a β‐turn, followed by and a helical element, to the N‐terminal region that presents only a β‐turn. We hypothesize that these two structured regions, by fluctuating independently in the lipid environment, may act in a coordinated fashion disrupting the yeast plasma membrane. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 88: 413–426, 2007.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

Gomesin (Gm) is a potent antimicrobial peptide isolated from the spider Acanthoscurria gomesiana. The two disulfide bridges Cys2,15 and Cys6,11 facilitate the folding of the molecule in a β‐hairpin structure, conferring on the peptide a high stability in human plasma. We report herein biological and structural features of new linear Gm analogues, obtained by combining the removal of both disulfide bridges and the incorporation of a D‐ or L‐proline. Regarding their biological properties, two analogues, namely, [D‐Thr2,6,11,15, Pro9]‐D‐Gm and [Thr2,6,11,15, D‐Pro9]‐Gm, are as potent as Gm against Candida albicans and only fourfold less against Staphylococcus aureus and Escherichia coli. In addition, at 100 μM they are approximately threefold less hemolytic than Gm. The best therapeutic indices were found for [D‐Thr2,6,11,15, Pro9]‐D‐Gm and for [(Des‐pGlu1, ‐Thr2, ‐Arg3), Thr6,11,15, D‐Pro9]‐Gm with a 32‐fold increase of their activity against bacteria, and from 128‐ to 512‐fold against yeast when compared with Gm. Regarding the stability, [D‐Thr2,6,11,15, Pro9]‐D‐Gm appeared to be the most resistant in human serum, along with [D‐Thr2,6,11,15, Pro8]‐D‐Gm and [Thr2,6,11,15, D‐Arg4,16, D‐Pro9]‐Gm. When evaluating their conformation by CD spectroscopy in sodium dodecyl sulfate (SDS), most linear analogues display β‐conformation characteristics. Moreover, considering its high therapeutic index and stability in serum, [D‐Thr2,6,11,15, Pro9]‐D‐Gm was further analyzed by NMR spectroscopy. 1H NMR experiments in SDS micelles demonstrated that [D‐Thr2,6,11,15, Pro9]‐D‐Gm presents a conformation very similar to that of Gm. In our search for Gm analogues with enhanced potential for drug development, we demonstrated that designing cysteine‐free analogues can improve the therapeutic index of Gm derivatives. © 2006 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 85:386‐400, 2007.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

The widely used programs BLAST (in this article, ‘BLAST’ includes both the National Center for Biotechnology Information [NCBI] BLAST® and the Washington University version WU BLAST) and FASTA for similarity searches in nucleotide and protein databases usually result in copious output. However, when large query sets are used, human inspection rapidly becomes impractical. BioParser is a Perl program for parsing BLAST and FASTA reports. Making extensive use of the BioPerl toolkit, the program filters, stores and returns components of these reports in either ASCII or HTML format. BioParser is also capable of automatically feeding a local MySQL® database with the parsed information, allowing subsequent filtering of hits and/or alignments with specific attributes. For this reason, BioParser is a valuable tool for large-scale similarity analyses by improving the access to the information present in BLAST or FASTA reports, facilitating extraction of useful information of large sets of sequence alignments, and allowing for easy handling and processing of the data.

Gomesin is an antimicrobial peptide isolated from hemocytes of the Brazilian spider Acanthoscurria gomesiana that contains two disulfide bridges Cys2‐15/Cys6‐11 and presents a β‐hairpin structure. To investigate the role of the disulfide bridges on gomesin conformation, bioactivities, and serum stability, structure–activity relationship (SAR) studies were conducted. Initially, gomesin and variants lacking one or both disulfide bridges were synthesized. CD studies showed that the gomesin structure is very rigid independently of the solvent environment. On the other hand, the linearized analogues adopted secondary structures according to the environment, while the monocyclic disulfide‐bridged peptides had a tendency to adopt a turn structure. The absence of one or both bridges resulted in a decrease in the antimicrobial and hemolytic activities. In addition, serum stability studies revealed that, contrasting to gomesin that was stable even after 48 h of incubation, the linearized analogues were rapidly degraded. The replacement of the disulfide bounds by lactam bridges led to monocyclic and bicyclic compounds. SAR studies indicated that the monocyclic lactam‐bridged analogues tend to assume a α‐helical structure being less potent, hemolytic, and serum stable than the wild‐type gomesin. On the other hand, the bicyclic lactam/disulfide‐bridged analogues displayed a similar conformation and degradation kinetics identical to gomesin. However, the antimicrobial activity appeared to be dependent on the lactam bridge position and size. These findings indicated that (i) the secondary structure plays a pivotal role for the full activity of gomesin; (ii) the antimicrobial and hemolytic activities of gomesin are correlated events; (iii) while at least one of the disulfide bridges is needed for the maintenance of a significant antimicrobial activity of gomesin, both bridges are required for high serum stability and optimal conformation; and finally (iv) the best analogue obtained was the bicyclo (2‐15,6‐11)[Glu2, Cys6,11, Lys15]‐Gm since it is as stable and potent as gomesin. © 2005 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 84: 205–218, 2006This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

ABSTRACTIt has not been possible to distinguish different strains of Mycobacterium lepraeaccording to their genetic sequence. However, the genome contains several variable-number tandem repeats (VNTR), which have been used effectively in strain typing of other bacteria. To determine their suitability for differentiating M. leprae, we developed PCR systems to amplify 5 different VNTR loci and examined a battery of 12 M. lepraestrains derived from patients in different regions of the United States, Brazil, Mexico, and the Philippines, as well as from wild armadillos and a sooty mangabey monkey. We found diversity at four VNTR (D = 0.74), but one system (C16G8) failed to yield reproducible results. Alleles for the GAA VNTR varied in length from 10 to 16 copies, those for AT17varied in length from 10 to 15 copies, those for GTA varied in length from 9 to 12 copies, and those for TA18varied in length from 13 to 20 copies. Relatively little variation was seen with interspecies transfer of bacilli or during short-term passage of strains in nude mice or armadillos. The TA18locus was more polymorphic than other VNTR, and genotypic variation was more common after long-term expansion in armadillos. Most strain genotypes remained fairly stable in passage, but strain Thai-53 showed remarkable variability. Statistical cluster analysis segregated strains and passage samples appropriately but did not reveal any particular genotype associable with different regions or hosts of origin. VNTR polymorphisms can be used effectively to discriminate M. lepraestrains. Inclusion of additional loci and other elements will likely lead to a robust typing system that can be used in community-based epidemiological studies and select clinical applications.

A combined resin solvation-peptide chain motion and kinetics of coupling reaction approach was applied to monitor details of the synthesis of TM-34, a 34-residue transmembrane segment of the bradykinin receptor. The dynamics of resin-bound peptide fragments attached to a stable free radical amino acid were examined by EPR spectroscopy. In agreement with an abrupt decrease (from 83 to 43%) in peptide purity occurring in the 12−16 region when DMF was used, a much more strongly immobilized chain population was detected, especially at the 12-mer stage. Conversely, faster couplings and improved synthesis were observed in 20% DMSO/NMP, probably due to the higher chain mobility in this mixed solvent. In addition, findings relating to solvation of peptide resins seemed to corroborate the previously advanced proposition that the 1:1 sum of electron acceptor and electron donor properties of a solvent can be considered to be an alternative and more appropriate parameter for its polarity. (© Wiley-VCH Verlag GmbH, 69451 Weinheim, Germany, 2002)

We give a description of Galilean particles in terms of geometric quantization,a geometric correspondence from quantum states in the preceding sense to wavefunctions in the ordinary quantum mechanical sense, and explicit computationsin some cases that lead to Schrödinger and Pauli equations.

Antifungal and antibacterial activities were detected in the hemolymph and gut contents of the cattle tick, Boophilus microplus. A peptide with antibacterial activity from the tick gut contents was purified to homogeneity by reversed-phase chromatography. The molecular mass of the purified peptide was 3,205.7 Da, measured by matrix-assisted laser desorption/ionization mass spectrometry. The amino acid sequence was obtained by Edman degradation and showed that the peptide was identical to a fragment of the bovine α-hemoglobin. A synthetic peptide based on the sequence obtained showed characterization data identical to those of the isolated material, confirming its structure. The synthetic peptide was active in micromolar concentrations against Gram-positive bacteria and fungi. These data led us to conclude that the antibacterial activity detected in tick gut contents is the result of enzymatic processing of a host protein, hemoglobin. This activity may be used by ticks as a defense against microorganisms.

Abstract: A geometrical way is described to associate quantum states in the sense of geometric quantization to wave functions in the quantum mechanical sense for each relativistic elementary particle. Explicit computations are made in a number of cases: Klein-Gordon and Dirac equations, neutrino and antineutrino Weyl equations, and very general cases of massive and massless particles of arbitrary spin. In this later case one is led in a canonical way to Penrose wave equations.

Background:Some of the most frequent manifestations of heart involvement in human immunodeficiency virus (HIV) infection include right and left ventricular dysfunction. The pathogenesis remains obscure.

Developments in information technology are bringing about fundamental changes in the ways in which users get access to primary documents, and give rise to some interesting paradoxes.

A combination of immunosuppressants may improve outcomes due to the synergistic effect of their different action mechanisms. Currently, there is no consensus regarding the best immunosuppressive protocol after liver transplantation. This review aimed to evaluate the effectiveness and safety of tacrolimus associated with mycophenolate mofetil (MMF) in patients undergoing liver transplantation. We performed a systematic review and meta-analysis of randomized clinical trials. Eight randomized trials were included. The proportion of patients with at least one adverse event related to the immunosuppression scheme with tacrolimus associated with MMF was 39.9%. The tacrolimus with MMF immunosuppression regimen was superior in preventing acute cellular rejection compared with that of tacrolimus alone (risk difference [RD]=-0.11; p=0.001). The tacrolimus plus MMF regimen showed no difference in the risk of adverse events compared to that of tacrolimus alone (RD=0.7; p=0.66) and cyclosporine plus MMF (RD=-0.7; p=0.37). Patients undergoing liver transplantation who received tacrolimus plus MMF had similar adverse events when compared to patients receiving other evaluated immunosuppressive regimens and had a lower risk of acute rejection than those receiving in the monodrug tacrolimus regimen.

Myocardial infarction is the foremost cause of death globally, and progression to heart failure is proportional to infarct size. Prevention of cardiomyocyte loss is vital to halt cardiac damage. Our previous data show that intramyocardial injection of cardiac progenitors (CPC) at time of infarct, results in remuscularization and scar size reduction. However, myogenic progenitors are poorly retained, hence paracrine signals might explain the observed improvement of cardiac function. Here paracrine effects of cardiac progenitors on human pluripotent stem cell derived cardiomyocytes (hPSCM) were investigated.To demonstrate whether CPC-conditioned media injections adequately prevent cardiomyocyte death, permanent ligation was performed in mice as a proof of principle in vivo. We then established in vitro systems to analyse the CPC-secretome effects in hPSCM. Conditioned medium injected into the infarct border zone at the time of coronary artery ligation reduced the proportion of apoptotic (TUNEL+) cardiomyocytes from 36.5%?7.8 to 9.8%?3.5 (n=6). Transwell coculture assays and conditioned medium experiments demonstrated that the secretome from mouse CPCs suppresses death from menadione induced-stress in hPSCM (DRAQ7) from 47.2%?2.9 to 2.7%?0.9 (n=9). Mechanistically, cardiomyocyte protection involved reducing apoptosis from 53.2%?5.1 to 12.6%?2.5 (n=9), preserving mitochondrial membrane potential (TMRM) from 26.4%?9.2 to 58.6%?3 (n=6) and inhibiting reactive oxygen species (CellROX) from 55%?6.3 to 12.6?2.1 (n=12). Functional investigations demonstrated that CPC-conditioned media preserve hPSCM beating frequency, calcium cycling amplitude (Fura-4F), and action potential characteristics (Di-4-ANEPPS, FluoVolt) following oxidative stress. Protection is exosome-independent and mediated by thermolabile molecules more than 3kDa in size. In summary, hPSCM were employed as a target platform for paracrine protection establishing their versatility to investigate biological signals that augment human cardiac muscle survival. CPC-secreted mediators impinge on apoptosis, preserve mitochondrial polarisation, suppress reactive oxygen content and inhibit loss of functional features in hPSCM exposed to oxidative stress.

The British Heart Foundation (BHF) annual image competition, Reflections of Research, provides a glimpse into the cutting-edge research that the BHF funds

In Brazil, pastures for cattle ranching are being established in areas that were previously forested. To investigate some consequences of this change in land use we measured fluxes of CO2 and water vapour over a typical pasture, dominated by the introduced C4 grass Brachiaria brizantha. In addition, we compared the CO2, water vapour fluxes and canopy stomatal conductances observed with those obtained simultaneously over a nearby undisturbed rain forest. Measurements were made near the end of the wet season under conditions of ample soil moisture. Leaf area index of the pasture was 3.9. The pasture had a lower canopy stomatal conductance than the forest (typically 0.2–0.3 mol m-2 s-1 versus 0.4–0.9 mol m-2 s-1 at high photon irradiance) and was less responsive to the canopy-to-air vapour pressure difference. As a consequence of these lower canopy stomatal conductances, the pasture used much less water than the forest with average values over the period examined being 153 mol H2O m-2 d-1 and 249 mol H2O m-2 d-1 for pasture and forest respectively (2.74 and 4.48 mm d-1 respectively). This was also reflected by differing fractions of the absorbed energy being dissipated as evaporation. This proportion was typically 0.56 for the pasture and 0.74 for the forest. After allowing for soil and plant respiration, average daily photosynthetic rates were 0.67 mol C m-2 d-1 for the pasture and 0.57 mol C m-2 d-1for the forest (8.0 and 6.8 g C m-2 d-1, respectively). Thus, despite an appreciably lower rate of water use the pasture assimilated more carbon on a daily basis. Nevertheless, Brachiaria displayed a somewhat lower rate of photosynthesis than expected for a C4 grass, perhaps because of a low nutrient status. Indeed, at low and medium photon irradiance the pasture and forest showed remarkably similar photosynthetic performance. There was, however, less tendency for CO2 assimilation rates of the pasture canopy to saturate at high photon irradiance. The respiratory fluxes from the two ecosystems at night were quite similar, 6–8 µmol m-2 s-1. The ratio of intercellular CO2 concentration to ambient CO2 concentration was usually 0.4 to 0.6 for the pasture, a range which is higher than that often reported for C4 plants but possibly not unusual for tropical grasses in their natural environment.