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Biopharmaceutical product and process development do not yet take advantage of predictive computational modeling to nearly the degree seen in industries based on smaller molecules. To assess and advance progress in this area, spirited coopetition (mutually beneficial collaboration between competitors) was successfully used to motivate industrial scientists to develop, share, and compare data and methods which would normally have remained confidential. The first "Highland Games" competition was held in conjunction with the October 2018 Recovery of Biological Products Conference in Ashville, NC, with the goal of benchmarking and assessment of the ability to predict development-related properties of six antibodies from their amino acid sequences alone. Predictions included purification-influencing properties such as isoelectric point and protein A elution pH, and biophysical properties such as stability and viscosity at very high concentrations. Essential contributions were made by a large variety of individuals, including companies which consented to provide antibody amino acid sequences and test materials, volunteers who undertook the preparation and experimental characterization of these materials, and prediction teams who attempted to predict antibody properties from sequence alone. Best practices were identified and shared, and areas in which the community excels at making predictions were identified, as well as areas presenting opportunities for considerable improvement. Predictions of isoelectric point and protein A elution pH were especially good with all-prediction average errors of 0.2 and 1.6 pH unit, respectively, while predictions of some other properties were notably less good. This manuscript presents the events, methods, and results of the competition, and can serve as a tutorial and as a reference for in-house benchmarking by others. Organizations vary in their policies concerning disclosure of methods, but most managements were very cooperative with the Highland Games exercise, and considerable insight into common and best practices is available from the contributed methods. The accumulated data set will serve as a benchmarking tool for further development of in silico prediction tools.

The progression of the human immunodeficiency virus (HIV) infection to acquired immunodeficiency syndrome (AIDS) can be efficiently interrupted by antiretroviral therapy (ART). However, even successfully treated HIV-infected individuals are prone to develop non-AIDS-related diseases that affect the metabolism and several organs and systems. Biomarkers that predict the occurrence of comorbidities may help develop preventive measures. Current research shows that CD4+ T cell counts and viral load do not predict the development of non-AIDS-related diseases. The CD4/CD8 ratio has been indicated as a suitable marker of persistent immune dysfunction and the occurrence of non-AIDS-related events in treated HIV-positive patients. In this study, we explored the relationship between CD4/CD8 ratios, comorbidities, and aging in ART-treated HIV patients on viral suppression. We collected and evaluated data from 352 HIV-positive adults who were virologically suppressed (

The Third Modeling Workshop focusing on bioprocess modeling was held in Kenilworth, NJ in May 2019. A summary of these Workshop proceedings is captured in this manuscript. Modeling is an active area of research within the biotechnology community, and there is a critical need to assess the current state and opportunities for continued investment to realize the full potential of models, including resource and time savings. Beyond individual presentations and topics of novel interest, a substantial portion of the Workshop was devoted toward group discussions of current states and future directions in modeling fields. All scales of modeling, from biophysical models at the molecular level and up through large scale facility and plant modeling, were considered in these discussions and are summarized in the manuscript. Model life cycle management from model development to implementation and sustainment are also considered for different stages of clinical development and commercial production. The manuscript provides a comprehensive overview of bioprocess modeling while suggesting an ideal future state with standardized approaches aligned across the industry.

The Brazilian state of Goias, untouched by spotted fever (SF) until 2012, has subsequently reported cases of the disease in several regions. This study aimed to survey the diversity of potential vectors and rickettsia in areas of Goias under environmental surveillance or case investigation for SF. Collected specimens were assayed with molecular biology technology using DNA extraction, amplification and sequencing of fragments of the genes gltA, ompA, ompB and sca4 to detect rickettsia in ticks and fleas. Amplification of cytochrome oxidase subunit II and 16S rRNA was performed to assist tick identification. Rickettsia felis (Rickettsiales: Rickettsiaceae) was found in Ctenocephalides felis (Bouche, 1835) (Siphonaptera: Pulicidae). Rickettsia bellii was found in Amblyomma rotundatum Koch, 1844 (Ixodida: Ixodidae) and in Amblyomma cajennense sensu lato. Rickettsia sp. strain NOD was found in Amblyomma nodosum Neumann, 1899. Of the Amblyomma cajennense complex, Amblyomma sculptum Berlese, 1888 was confirmed in the northern, northeast, midwest and southeast regions of Goias, whereas Amblyomma cajennense sensu stricto (Fabricius, 1787) was found only in the northern region of the state. Amblyomma dubitatum Neumann, 1899 associated with a species of the A. cajennense complex was the most common epidemiological finding, although Rickettsia rickettsii was not detected. This is the first report of Rickettsia sp. strain NOD in Goias.

Aim The present study aimed to characterize the prevalence of anemia and iron deficiency in older Portuguese adults, and to compare it with the prevalence in younger individuals. Methods A population-based, cross-sectional study (EMPIRE study) enrolling a representative sample of 6267 adults aged

Biosurfactants are microbial secondary metabolites. The most studied are rhamnolipids, which decrease the surface tension and have emulsifying capacity. In this study, the production of biosurfactants, with emphasis on rhamnolipids, and diesel oil degradation by 18 strains of bacteria isolated from waste landfill soil contaminated by petroleum was analyzed. Among the studied bacteria, gram-positive endospore forming rods (39%), gram positive rods without endospores (17%), and gram-negative rods (44%) were found. The following methods were used to test for biosurfactant production: oil spreading, emulsification, and hemolytic activity. All strains showed the ability to disperse the diesel oil, while 77% and 44% of the strains showed hemolysis and emulsification of diesel oil, respectively. Rhamnolipids production was observed in four strains that were classified on the basis of the 16S rRNA sequences as Pseudomonas aeruginosa. Only those strains showed the rhlAB gene involved in rhamnolipids synthesis, and antibacterial activity against Escherichia coli, P. aeruginosa, Staphylococcus aureus, Bacillus cereus, Erwinia carotovora, and Ralstonia solanacearum. The highest production of rhamnolipids was 565.7 mg/L observed in mineral medium containing olive oil (pH 8). With regard to the capacity to degrade diesel oil, it was observed that 7 strains were positive in reduction of the dye 2,6-dichlorophenolindophenol (2,6-DCPIP) while 16 had the gene alkane mono-oxygenase (alkB), and the producers of rhamnolipids were positive in both tests. Several bacterial strains have shown high potential to be explored further for bioremediation purposes due to their simultaneous ability to emulsify, disperse, and degrade diesel oil. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:262-270, 2016.

Three copper(II) complexes, [Cu2(O2CC4H3O)4(H2O)2]·H2O (1), {[Cu2(O2CC4H3O)4(bipy)2(H2O)2]·2H2O}n(2), and {[Cu(bpp)2(H2O)](O2CC4H3O)2·7H2O} (3) [O2CC4H3O = 2-furanocarboxylate anion, bipy = 4, 4′-bipyridine, and bpp = 1, 3-bis(4-pyridyl)propane] were synthesized and their crystal and molecular structures were determined by single-crystal X-ray diffraction analysis. Compound 1 consists on bimetallic tetracarboxylate units exhibiting a common paddle wheel structure. The paddle wheel units are connected by hydrogen bonding along the crystallographic b axis and π–π stacking interactions between the furano rings along the crystallographic c axis, giving rise to a 2D supramolecular array. Compounds 2 and 3 were made up in the presence of 4, 4′-bipyridine and 1, 3-bis(4-pyridyl)propane ligands, respectively, to produce 1D polymeric chains by metal-ligand interactions. The presence of other weak non-covalent interactions in these compounds extends the systems dimensionality to form 3D supramolecular networks.

Two series of novel cationic gemini surfactants based on the amino acid serine have been synthesized. These compounds contain long alkyl chains linked to the nitrogen atoms of the amino acid residues, as well as spacers interconnecting two carboxylic acid groups, through amide or ester bonds. The most efficient synthetic pathway was established; it involves the introduction of the spacers into the N,N-dialkylated monomeric precursors by peptide condensation methods with diamines or diols, with subsequent methylation and deprotection to yield the final target surfactants. The effects of molecular structure, type of spacer linkage, and spacer length on the interfacial and cytotoxic properties of these gemini surfactants are reported and discussed; studies on gemini surfactants with amine linkages and other relevant homologous compounds (conventional bis-quat gemini and monomeric surfactants) are also included. Overall, it is shown that cationic serine-based gemini surfactants have enhanced interfacial properties and low cytotoxicities, offering potential use in technical and biological applications.

The present study aimed to investigate leucocyte responses to inflammation as well as some innate immune parameters of Senegalese sole, Solea senegalensis, following challenge with two strains of Photobacterium damselae subsp. piscicida belonging to the European and Japanese clones described for this bacterium. Pathogenicity assays were performed to assess the virulence of each Photobacterium damselae subsp. piscicida strain for sole. Subsequently, fish were intraperitoneally injected with phosphate-buffered saline (control) or two concentrations (2 × 10² and 2 × 10⁶ CFU mL⁻¹) of each bacterial strain and sampled after 6 and 24 h. Results showed that the European isolate induces a higher degree of response than the Japanese strain. While blood neutrophilia and monocytosis correlated well with the increase in neutrophil and macrophage numbers in the peritoneal cavity, fish infected with the European isolate presented higher peritoneal cell numbers than fish challenged with the Japanese strain. In addition, alternative complement pathway activity and respiratory burst of head kidney leucocytes increased significantly in fish infected with the European isolate. The enhanced innate immune response displayed by Senegalese sole challenged with the European isolate is probably due to the higher degree of virulence presented by this Photobacterium damselae subsp. piscicida strain.

The synthesis of novel cationic gemini surfactants based on serine, with long lipophilic alkyl chains and a spacer linked to the nitrogen atom of the amino acid residue by amine linkages, is described. The most efficient synthetic pathway involves introduction of the spacer into the monomeric precursors, N-alkyl derivatives, by reductive amination of dialdehydes followed by methylation and deprotection. Characterization of the basic micellization properties of the new compounds was carried out by tensiometry and conductimetry. These surfactants present enhanced interfacial properties compared to the monomeric analogues, and show improved performance, namely lower critical micelle concentration (cmc), lower surface tension at cmc and increased micellar ionization, with respect to conventional bis-quaternary ammonium salts (bis-quats).

Background and purpose: The present study reports on the preparation and testing of a sustained delivery system for the immunomodulatory peptide P10 aimed at reducing the in vivo degradation of the peptide and the amount required to elicit a protective immune response against paracoccidioidomycosis.

The ability to process high-concentration monoclonal antibody solutions (> 10 g/L) through small-pore membranes typically used for virus removal can improve current antibody purification processes by eliminating the need for feed stream dilution, and by reducing filter area, cycle-time, and costs. In this work, we present the screening of virus filters of varying configurations and materials of construction using MAb solutions with a concentration range of 4-20 g/L. For our MAbs of interest-two different humanized IgG1s-flux decay was not observed up to a filter loading of 200 L/m(2) with a regenerated cellulose hollow fiber virus removal filter. In contrast, PVDF and PES flat sheet disc membranes were plugged by solutions of these same MAbs with concentrations >4 g/L well before 50 L/m(2). These results were obtained with purified feed streams containing

Heart failure (HF) costs are largely due to hospitalization. The validity of a death/discharge diagnosis of HF (DDHF) is largely unknown. The authors assessed the validity of DDHF and the impact of misdiagnosing. The case notes of patients consecutively admitted to a medical department between January and June 2001 were reviewed. Cases with DDHF or cardiovascular diseases, potential precursors of HF (PPHF), were included. The diagnosis of HF (European Society of Cardiology guidelines) was classified as definite, possible, or miscoded. Of the 1038 patients admitted, 234 were enrolled: 157 with DDHF and 77 with PPHF. One hundred eighty patients had a definite diagnosis of HF. Of the 157 diagnoses coded as definite HF, 130 were correct, 21 had possible HF, and 6 were miscoded. Of the 77 patients classified as having PPHF, 38 had definite HF. The accuracy of the DDHF diagnosis was 72.2%: 21.1% were underdiagnosed and 8.3% overdiagnosed. DDHF failed to capture many HF admissions and therefore alone underestimates the prevalence, burden, and costs of the syndrome.

The elemental concentration in buried human bones is studied by energy dispersive x-ray fluorescence. Several cortical and trabecular bones: skulls, peroneus, mandibles, tibias, femurs, ribs, vertebras and omoplates, belonging to a population dating to the Middle Ages were analysed. The purpose of this study is to compare the behaviour of the different structures of the several bones during the diagenetic process affecting bone during burial. Quantitative analysis was performed for K, Ca, Zn, Sr, Ti, Mn, Fe and Pb and cluster analysis was achieved considering the elemental concentrations for the different bones. There is a clear distinction for bone association regarding Ca, Zn, Sr and K, Mn, Fe, Ti and Pb in the several bones. These results do not allow a clear categorization into cortical and trabecular bones for some of the samples studied, as far as distinguishing between the remains of the original bone and the environmental contamination from the surrounding soil. Copyright © 2007 John Wiley & Sons, Ltd.

Digenean metacercariae of 0 year group common sole Solea solea (n = 70) were more abundant in the embayed nursery of the Pertuis Charentais than in the Tagus estuary nursery. Macroparasite assemblages of adult sole (n = 119) displayed only one species in common between the Bay of Biscay and the Portuguese coast continental shelves. These data highlighted the potential use of macroparasites as biological tags in various aspects of common sole ecology. (C) 2007 The Authors.

Macro- and microelement contents of five medicinal plants (Taraxacum officinale Weber, Eucalyptus globulus Labill, Plantago lanceolata L., Matricaria chamomilla L. and Mentha piperita L.) and their infusions were evaluated by the combined use of x-ray fluorescence (WDXRF and EDXRF, bulk raw plants) and inductively coupled plasma (ICP-MS and ICP-AES, infusions) techniques. The analytical methods allow the determination of 17 elements (Na, Mg, Al, Si, P, S, K, Ca, Ti, Mn, Fe, Cu, Zn, As, Rb, Sr, and Pb) both in plants and in the infusions. The use of XRF techniques offer a good multielemental approach for the rapid quality control of bulk raw plant materials whereas ICP techniques are well suited for the analytical control of infusions in order to ascertain the nutritional role of medicinal plants and the daily dietary intake. Copyright © 2005 John Wiley & Sons, Ltd.

The purpose of the present work was to investigate whether the distribution of trace elements in human teeth of the Neolithic period in Portugal can reflect environmental conditions, dietary habits and uptake of some elements from the surrounding soil. An X-ray fluorescence set-up with microprobe capabilities, installed at the LURE synchrotron (France), was used for elemental analysis of teeth. The resolution of the synchrotron microprobe was 100 μm and the energy of the incident beam was 18 keV. The synchrotron microprobe concentration profiles for V, Mn, Fe, Cu, Zn, Sr and Pb in different regions of the tooth are discussed and correlated with the results obtained by cranio-facial studies. The Mn and Fe concentrations were much higher in Neolithic teeth than in those of present humans. This may suggest uptake of both elements from the soil. Low concentration levels of Sr were found, which can be correlated with dietary habits. Moreover, the Pb concentration was very low in all the teeth analysed, in agreement with the low pollution levels of this element in this population. All samples were analysed without any chemical preparation.

Arsenic and other heavy metal concentrations in post mortem soft tissues from a Portuguese King of the nineteenth century were studied by energy-dispersive x-ray fluorescence analysis. This work is an attempt to clarify the strange death of the King, through the analysis of his post mortem soft tissue remains, which were kept inside a Chinese porcelain container since his death in 1826. The container was put in a wooden coffin and buried underground in a chapel of the St Vicente de Fora Monastery in Lisbon. Quantitative elemental analysis of the remains showed very high concentrations of arsenic and lead, which on average correspond to a few hundred times the mean normal values for contemporary human soft tissues. Increased levels of Cu were found, and also small enrichments for Fe and Zn, probably due to either enrichment during the 175 years or contamination by direct intake from the container, or both. The results obtained are consistent with acute arsenic poisoning, reinforcing the conclusions from the medical anatomical–pathological report, based on the analysis of the fragments by scanning electron microscopy. Copyright © 2002 John Wiley & Sons, Ltd.

X-ray fluorescence spectrometry was used for the quantitative analysis of healthy liver, brain and kidney tissues and the corresponding tissues from subjects suffering from liver cirrhosis. All samples were collected post-mortem and frozen until analysis. A sample of each tissue for the same individual was collected, in order to find any correlation between the elemental concentrations in the different tissues. Age, sex, cause of death and specific diseases were registered. All samples were studied without any chemical treatment. Concentrations of minor and trace elements in each tissue were obtained for 14 elements. Three groups of elements were considered. The first was K and Ca, present in relatively high concentrations in all tissues and known as normal constituents of all living matter. They are essential to cellular metabolism. The second group was Mn, Fe, Cu, Zn, Se, Br, Rb and Sr, which are also essential elements, but may induce toxic effects if present in high concentrations. There is a narrow range between beneficial and toxic concentrations for these elements. The third group was the toxic elements Co, Ni, As and Pb, which normally induce pathological disorders in the organism, even in small concentrations. This work is a contribution to the better understanding of trace element accumulation in human liver, kidney and brain and the possible correlation between abnormal concentrations of some elements in tissues of subjects suffering from cirrhosis. A very important factor is also the relative concentration of the elements with respect to one another and the correlation between the elemental content in the different tissues. Very low concentrations for Ni, As and Sr were found in the samples analysed, at the level of the detection limit. Higher concentrations of Co, Fe and Pb and lower values of Zn and Se were found in liver cirrhosis samples compared with healthy tissues. Brain samples from subjects affected with cirrhosis showed enhanced levels of Pb. In kidney, increased amounts of Zn were found in tissues from subjects with cirrhosis. Copyright © 2001 John Wiley & Sons, Ltd.

Two patients with adenoid cystic carcinoma of Bartholin's gland and a review of the relevant literature are presented. With the inclusion of these two patients, there are now 24 cases reported. Both patients had large vulvar masses with a short clinical history, and several local tumor recurrences within the first 21/2 years after radical vulvectomy. The characteristic cribriform pattern and perineural involvement in addition to vascular invasion were present in the pathological material. No metastases were found in the inguino-femoral lymph nodes removed. Both patients are alive, without evidence of local recurrence but with lung metastases. A chemotherapeutic treatment scheme (adriamycin and cyclophosphamide) is underway but it is too early to evaluate its results.