Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi.

Polyketides with the isochroman-3-one pharmacophore are rare among fungal natural products as their biosynthesis requires an unorthodox S-type aromatic ring cyclization. Genome mining uncovered a conserved gene cluster in select leotiomycetous fungi that encodes the biosynthesis of cytosporones, including isochroman-3-one congeners. Combinatorial biosynthesis in total biosynthetic and biocatalytic formats in Saccharomyces cerevisiae and in vitro reconstitution of key reactions with purified enzymes revealed how cytosporone structural and bioactivity diversity is generated. The S-type acyl dihydroxyphenylacetic acid (ADA) core of cytosporones is assembled by a collaborating polyketide synthase pair. Thioesterase domain-catalyzed transesterification releases ADA esters, some of which are known Nur77 modulators. Alternatively, hydrolytic release allows C6 hydroxylation by a flavin-dependent monooxygenase, yielding a trihydroxybenzene moiety. Reduction of the C9 carbonyl by a short chain dehydrogenase/reductase initiates isochroman-3-one formation, affording cytosporones with cytotoxic and antimicrobial activity. Enoyl di- or trihydroxyphenylacetic acids are generated as shunt products, while isocroman-3,4-diones are formed by autoxidation. The cytosporone pathway offers novel polyketide biosynthetic enzymes for combinatorial synthetic biology to advance the production of "unnatural" natural products for drug discovery.

[First Case report of Lawsonella clevelandensis Infection in Latin America and literature review]. / Reporte del primer caso de infección por Lawsonella clevelandensis en Latinoamérica y revisión de literatura.

Lawsonella clevelandensis is a gram-positive anaerobic bacillus, partially acid-fast, belonging to the order Corynebacteriales and is considered commensal of the skin and oral cavity. In recent years, few cases of infection by this microorganism have been reported, generally in immunosuppressed patients, with the most reported clinical presentation being the generation of abscesses. The isolation of this bacterium is difficult since it does not grow in traditional cultures, so its identification usually requires the use of molecular biology. We present the case of a 29-year-old patient with a kidney transplant and progressive worsening of kidney function in the context of a pauci-immune vasculitis which required greater immunosuppression. During the investigation, it was identified an asymptomatic leukocyturia, with a negative urine culture, for which six urine smear microscopy tests were performed of which 3 were positive, but with a negative Koch culture. Sequencing of the gene that codes for the 16s RNA of the bacterial ribosome was performed from a urine sample, obtaining a sequence compatible with Lawsonella clevelandensis. After the result, therapy with amoxicillin-clavulanic acid was indicated with resolution of the leukocyturia and negative smear microscopy, and immunosuppression could be indicated without inconvenience. To the authors' knowledge, this is the twelfth case of Lawsonella clevelandensis infection described in the medical literature in the world and the first to report the isolation of the pathogen in urine.

[Managing Superwarfarin Poisoning: A Challenging Case]. / Relato de caso de intoxicación con superwarfarinas: un desafío terapéutico.

Human intoxication by long-acting anticoagulant rodenticides, known as superwarfarins, causes coagulopathy that is difficult to manage. We present the case of a 42-year-old man who ingested a toxic dose of rodenticide in a suicide attempt, evolving with epistaxis, INR of 11.6, and needing hospitalization. For seven days, serial controls of coagulation tests were carried out, with optimization of different doses of Vitamin K supplementation. The case highlights this type of anticoagulant's potency and prolonged half-life (approximately six weeks), which requires regular clinical control and satisfactory treatment adherence.

Microstructure, Phase Evolution, and Chemical Behavior of CrCuFeNiTiAlx High Entropy Alloys Processed by Mechanical Alloying.

High entropy alloys (HEAs) of the type CrCuFeNiTi-Alx were processed through mechanical alloying. The aluminum concentration was varied in the alloy, to determine its effect on the HEAs' microstructure, phase formation, and chemical behavior. X-ray diffraction studies performed on the pressureless sintered samples revealed the presence of structures composed of face centered cubic (FCC) and body centered cubic (BCC) solid-solution phases. Since the valences of the elements that form the alloy are different, a nearly stoichiometric compound was obtained, increasing the final entropy of the alloy. The aluminum was partly responsible for this situation, which also favored transforming part of the FCC phase into BCC phase on the sintered bodies. X-ray diffraction also indicated the formation of different compounds with the alloy's metals. Bulk samples exhibited microstructures with different phases. The presence of these phases and the results of the chemical analyses revealed the formation of alloying elements that, in turn, formed a solid solution and, consequently, had a high entropy. From the corrosion tests, it could be concluded that the samples with a lower aluminum content were the most resistant to corrosion.

Characterization of the perioperative changes of exosomal immune-related cytokines induced by prostatectomy in early-stage prostate cancer patients.

BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are relevant in prostate cancer microenvironment collaborating in tumor development. The main tumor marker used in this disease, prostate-specific antigen (PSA), does not provide information related to this tumor microenvironment. Cancer cells secrete exosomes carrying bioactive molecules contributing to MDSCs recruitment and induction. The aim of this study was to characterize the perioperative changes of exosomal cytokines relevant in MDSCs recruitment induced by prostatectomy in prostate cancer patients. METHODS: Blood was drawn from 26 early-stage prostate cancer patients before and after radical prostatectomy and from 16 healthy volunteers. Serum exosomes were separated by precipitation. Cytokines related with MDSC cell recruitment and activation CCL2, CXCL2, CXCL5, CXCL8, CXCL12, MIF, S100A9 and TGF-ß were measured in serum and serum-derived exosomes using immunometric assays. RESULTS: All cytokines were detected both in serum and exosomes, except for CXCL12, which was detected only in serum. Exosomes were enriched specially in MIF, TGF-ß and CXCL2. Presurgical MIF levels in exosomes correlated negatively with serum PSA. Also, presurgical TGF-ß decreased both in serum and exosomes as Gleason score rises. Patients presurgical exosomes had increased CCL2, CXCL5 and TGF-ß levels than exosomes from healthy controls. These differences were not observed when cytokines were analyzed in serum, except for TGF-ß. Cytokine levels of CCL2, CXCL5 decreased in patients' postsurgical exosomes, while TGF-ß further increased. On the contrary, S100A9 levels were lower in patients presurgical exosomes but increased after radical prostatectomy. CONCLUSIONS: Blood exosomal content in cytokines constitute an attractive source to evaluate MDSCs immunomodulators providing additional information related to tumor microenvironment in prostate cancer.

Bronchoscopist's perception of the quality of the single-use bronchoscope (Ambu aScope4™) in selected bronchoscopies: a multicenter study in 21 Spanish pulmonology services.

BACKGROUND: The disposable bronchoscope is an excellent alternative to face the problem of SARS-CoV-2 and other cross infections, but the bronchoscopist's perception of its quality has not been evaluated. METHODS: To evaluate the quality of the Ambu-aScope4 disposable bronchoscope, we carried out a cross-sectional study in 21 Spanish pulmonology services. We use a standardized questionnaire completed by the bronchoscopists at the end of each bronchoscopy. The variables were described with absolute and relative frequencies, measures of central tendency and dispersion depending on their nature. The existence of learning curves was evaluated by CUSUM analysis. RESULTS: The most frequent indications in 300 included bronchoscopies was bronchial aspiration in 69.3% and the median duration of these was 9.1 min. The route of entry was nasal in 47.2% and oral in 34.1%. The average score for ease of use, image, and aspiration quality was 80/100. All the planned techniques were performed in 94.9% and the bronchoscopist was satisfied in 96.6% of the bronchoscopies. They highlighted the portability and immediacy of the aScope4TM to start the procedure in 99.3%, the possibility of taking and storing images in 99.3%. The CUSUM analysis showed average scores > 70/100 from the first procedure and from the 9th procedure more than 80% of the scores exceeded the 80/100 score. CONCLUSIONS: The aScope4™ scored well for ease of use, imaging, and aspiration. We found a learning curve with excellent scores from the 9th procedure. Bronchoscopists highlighted its portability, immediacy of use and the possibility of taking and storing images.

Early diversification and permeable species boundaries in the Mediterranean firs.

BACKGROUND AND AIMS: Inferring the evolutionary relationships of species and their boundaries is critical in order to understand patterns of diversification and their historical drivers. Despite Abies (Pinaceae) being the second most diverse group of conifers, the evolutionary history of Circum-Mediterranean firs (CMFs) remains under debate. METHODS: We used restriction site-associated DNA sequencing (RAD-seq) on all proposed CMF taxa to investigate their phylogenetic relationships and taxonomic status. KEY RESULTS: Based on thousands of genome-wide single nucleotide polymorphisms (SNPs), we present here the first formal test of species delimitation, and the first fully resolved, complete species tree for CMFs. We discovered that all previously recognized taxa in the Mediterranean should be treated as independent species, with the exception of Abies tazaotana and Abies marocana. An unexpectedly early pulse of speciation in the Oligocene-Miocene boundary is here documented for the group, pre-dating previous hypotheses by millions of years, revealing a complex evolutionary history encompassing both ancient and recent gene flow between distant lineages. CONCLUSIONS: Our phylogenomic results contribute to shed light on conifers' diversification. Our efforts to resolve the CMF phylogenetic relationships help refine their taxonomy and our knowledge of their evolution.

[Social inequality in the use of screening tests in Mexico: an analysis of the national health surveys 2006 and 2012]. / Desigualdad social en México enel uso de servicios de tamizaje en adultos: un análisis de las encuestas nacionales de salud 2006 y 2012.

OBJECTIVE: To measure health inequality in the use of screen-ing services in adults from 20 to 59 years of age from the 2006 and 2012 national health and nutrition surveys. MATERIALS AND METHODS: dults (detection of diabetes, hypertension, breast cancer, cervical cancer and prostate cancer), the Kuznets index, the slope inequality index and the health concentration index were estimated. Considering as social indicators schooling, ethnicity, unemployment, socioeconomic level and type of health protection. RESULTS: The coverage of the five tests increased, but the inequality observed only decreased in the interventions in women; and in the case of the detection of prostate cancer it was increased. CONCLUSIONS: While it is important to monitor the performance of curative services, the challenge remains to ensure effective and equitable access to early diagnosis services.

OBJETIVO: Medir la desigualdad en el uso de servicios de tamizaje en adultos de 20 a 59 años, a partir de las encuestas nacionales de salud y nutrición 2006 y 2012. MATERIAL Y MÉTODOS: A partir de la selección de cinco indicadores de tamizaje en adultos (detección de diabetes, hipertensión y cánceres de mama, cérvicouterino y de próstata) se estimaron el índice de Kuznets, el índice de desigualdad de la pendiente y el índice de concentración de salud, considerando como indicadores sociales la escolaridad, etnicidad, desempleo, nivel socioeconómico y tipo de protección en salud. RESULTADOS: Las coberturas de las cinco pruebas se incrementaron, sin embargo, la desigualdad observada disminuyó únicamente en las intervenciones en mujeres; en el caso de la detección de cáncer de próstata se incrementó. CONCLUSIONES: Si bien es importante monitorear el desempeño de los servicios curativos, persiste el reto de asegurar el acceso efectivo y equitativo a servicios de diagnóstico temprano.

Prevalence analysis of urinary incontinence after radical prostatectomy and influential preoperative factors in a single institution.

AIMS: To assess prevalence of urinary incontinence (UI) after radical prostatectomy (RP) and to analyze which preoperative characteristics of the patients have influence on UI. METHODS: Between 2002 and 2012, 746 consecutive patients underwent RP for clinically localized prostate cancer. We defined UI according to International Continence Society (ICS) definition: "the complaint of any involuntary leakage of urine" after 12 months of recovery, international consultation on incontinence questionnaire (ICIQ-SF) and pads/day was collected too. Clinical features and magnetic resonance imaging measurements were assessed. A multivariable logistic regression model predicting incontinence were built-in after adjust by cofounding factors and bootstrapping. RESULTS: About 172 (23%) of the patients were classified as incontinent according to the ICS definition. The mean value of the ICIQ-SF was 10.87 (±4). 17.8% of patients use at least one pad/day, 11.9% use more than one pad/day. The preoperative factors independently influential in UI are: age [OR: 1.055; CI 95% (1.006-1.107), p = .028], urethral wall thickness [OR: 5.03; CI 95% (1.11-22.8), p = .036], history of transurethral resection of the prostate [OR: 6.13; CI 95% (1.86-20.18), p = .003] and membranous urethral length [OR: 0.173; CI 95% (0.046-0.64), p = .009]. The predictive accuracy of the model is 78.7% and the area under the curve (AUC) value 71.7%. CONCLUSIONS: Urinary incontinence after radical prostatectomy has different prevalence depending on the definition. Age, prior transurethral resection of the prostate (TURP), membranous urethral length (MUL) and urethral wall thickness (UWT) were risk factors.

[Behavior of influenza seasons in Mexico from 2010 to 2016: Analysis and prospective]. / Comportamiento de las temporadas de influenza en México de 2010 a 2016, análisis y prospectiva.

The influenza virus spreads rapidly through recurring seasonal outbreaks during the fall and winter. Our country has the Epidemiological Surveillance System for Influenza (SISVEFLU), in operation since 2006, which has records of 558 health units of Influenza. According to the information recorded in it, we can see that the 2010-2016 influenza seasons have a biannual behavior, and that in the 2010-2011, 2012-2013, and 2014-2015 seasons, the predominant viral subtype was A (H3N2), while in the 2011-2012, 2013-2014, and 2015-2016 seasons, the predominant subtype was A (H1N1) pdm09, which was associated with an increased number of influenza cases and deaths. It is expected that the 2016-2017 season will have predominance of subtype A (H3N2) and in 2017-2018 the expected will be subtype A (H1N1) pmd09. During the 2010-2016 seasons, 53.5% of cases of influenza were women; 77% had no history of vaccination, and 36% had one or more comorbidities. As for deaths, 55% was observed in males, 85% had not been vaccinated, and 71.5% had one or more comorbidities.

The alpha-galactosidase A p.Arg118Cys variant does not cause a Fabry disease phenotype: data from individual patients and family studies.

Lysosomal α-galactosidase A (α-Gal) is the enzyme deficient in Fabry disease (FD), an X-linked glycosphingolipidosis caused by pathogenic mutations affecting the GLA gene. The early-onset, multi-systemic FD classical phenotype is associated with absent or severe enzyme deficiency, as measured by in vitro assays, but patients with higher levels of residual α-Gal activity may have later-onset, more organ-restricted clinical presentations. A change in the codon 118 of the wild-type α-Gal sequence, replacing basic arginine by a potentially sulfhydryl-binding cysteine residue - GLA p.(Arg118Cys) -, has been recurrently described in large FD screening studies of high-risk patients. Although the Cys118 allele is associated with high residual α-Gal activity in vitro, it has been classified as a pathogenic mutation, mainly on the basis of theoretical arguments about the chemistry of the cysteine residue. However its pathogenicity has never been convincingly demonstrated by pathology criteria. We reviewed the clinical, biochemical and histopathology data obtained from 22 individuals of Portuguese and Spanish ancestry carrying the Cys118 allele, including 3 homozygous females. Cases were identified either on the differential diagnosis of possible FD manifestations and on case-finding studies (n=11; 4 males), or on unbiased cascade screening of probands' close relatives (n=11; 3 males). Overall, those data strongly suggest that the GLA p.(Arg118Cys) variant does not segregate with FD clinical phenotypes in a Mendelian fashion, but might be a modulator of the multifactorial risk of cerebrovascular disease. The Cys118 allelic frequency in healthy Portuguese adults (n=696) has been estimated as 0.001, therefore not qualifying for "rare" condition.

Suicide note and the psychological autopsy: Associated behavioral aspects.

When a death occurs under traumatic, ambiguous, unknown, or uncertain circumstances, a death of suicidal origin can be considered. On many occasions, a suicide note is found next to body that helps to clarify certain aspects needed for the investigation to elucidate whether the death is really due to suicide or other causes. There are several types of suicide note (farewell, instructions, accusation of others, request for forgiveness, justification of one’s own suicide) that can contribute to the study of the victim’s psychological state and the circumstances that led to death. There is no unanimously approved way to conduct the psychological autopsy, but there are protocols for obtaining relevant information and preparing the report.

Effects of tree architecture on pollen dispersal and mating patterns in Abies pinsapo Boiss. (Pinaceae).

Plant architecture is crucial to pollination and mating in wind-pollinated species. We investigated the effect of crown architecture on pollen dispersal, mating system and offspring quality, combining phenotypic and genotypic analyses in a low-density population of the endangered species Abies pinsapo. A total of 598 embryos from three relative crown height levels (bottom, middle and top) in five mother plants were genotyped using eleven nuclear microsatellite markers (nSSRs). Paternity analysis and mating system models were used to infer mating and pollen dispersal parameters. In addition, seeds were weighed (N = 16 110) and germinated (N = 736), and seedling vigour was measured to assess inbreeding depression. Overall, A. pinsapo shows a fat-tailed dispersal kernel, with an average pollen dispersal distance of 113-227 m, an immigration rate of 0.84-26.92%, and a number of effective pollen donors (Nep ) ranging between 3.5 and 11.9. We found an effect of tree height and relative crown height levels on mating parameters. A higher proportion of seeds with embryo (about 50%) and a higher rate of self-fertilization (about 60%) were found at the bottom level in comparison with the top level. Seed weight and seedling vigour are positively related. Nevertheless, no differences were found in seed weight or in seedling-related variables such as weight and length of aerial and subterranean parts among the different relative crown height levels, suggesting that seeds from the more strongly inbred bottom level are not affected by inbreeding depression. Our results point to vertical isotropy for outcross-pollen and they suggest that self-pollen may ensure fertilization when outcross-pollen is not available in low-density population.

Phylogeography of SW Mediterranean firs: different European origins for the North African Abies species.

The current distribution of Western Mediterranean Abies species is a result of complex geodynamic processes and climatic oscillations that occurred in the past. Abies sect. Piceaster offers a good study model to explore how geo-climatic oscillations might have influenced its expansion and diversification on both sides of the W Mediterranean basin. We investigated the genetic variation within and among nine populations from five Abies species by molecular markers with high and low mutation rates and contrasting inheritance (AFLP and cpSSR). Analyses revealed the opening of the Strait of Gibraltar as an effective barrier against gene flow between the Southern Iberian (A. pinsapo) and North African (A. marocana and A. tazaotana) firs. The A. pinsapo populations in Spain and likewise those of the A. marocana - A. tazaotana population complex were not differentiated, and no evidence was found to distinguish A. tazaotana at the species level. Diversification of Abies across North Africa could occur by way of at least two vicariant events from Europe, in the west, giving rise to the A. marocana - A. tazaotana complex, and in the east, giving A. numidica. Secondary contacts among species from Abies sect. Piceaster (A. pinsapo and A. numidica), and with A. alba (Abies sect. Abies) are also indicated. However, there is a closer relationship between the Algerian fir (A. numidica) and the North Mediterranean widespread A. alba, than with the Moroccan firs (A. marocana and A. tazaotana) or the Southern Iberian (A. pinsapo). We also discuss the distribution range of these taxa in its paleogeological and paleoclimatic context, and propose that part of the modern geography of the South-Western Mediterranean firs might be traced back to the Tertiary.

Radical prostatectomy. Detailed surgical margins. Prognostic value of multifocal involvement in pT2 (+).

OBJECTIVES: We intend to analyze the prognostic value of positive surgical margins depending on their number and location in pT2 patients. METHODS: We analyze 448 (34.3%) patients with positive surgical margins from a series of 1,310 T1-T2 patients treated with radical prostatectomy between 1989-2012. Of them 164 are pT2 (+). 119 (72.6% ) have unifocal affectation (41 (34.5%) unifocal in right lobe; 35 (29.4%) unifocal in left lobe, 40 (33.6%) unifocal in apex, 3 (2.5% ) unifocal proximal) and 45 (27.4%) multifocal involvement. RESULTS: Unifocal and multifocal pT2(+)patients have not evidenced significant differences in any of the clinicopathologic variables compared. However the BPFS at 5 and 10 years is significantly worse in the multifocal group, (p<0.000) In the BPFS multivariate study of 164 pT2(+ )influential variables are: multifocal involvement (HR: 3.4; 95%IC 1.7-6.9 p<0.000) and PSA (HR: 1.03; 95%IC 1.02-1.05 p<0.000), being PSA >15 ng/ml )HR: 3.7; 95%IC 2.1-6.6 p<0.000 ( the best cut-off point. Risk groups: Using the independent influence variables, the best model (using Cox models ) includes two risk groups: Group 1 (0 variables): They are pT2(+) with unifocal affectation and PSA<15 ng/ml, (63%). Their BPFS are 81±4% and 77±4% (5 and 10 years). Grupo 2 (1-2 variables): They are pT2 (+) with multifocal involvement, PSA> 15 ng/ml or both of them, (37%). Their BPFS are 46±6% and 26±7% (5 and 10 years). The BPFS differs significantly between the two groups (p<0.000). The Group 1 BPFS is similar to the pT2 (-) patients, (p:0.242). The Group 2 BPFS is similar to the pT3(+) patients, (p:0.637). The model explained significantly better the BPFS than any of the individual variables analyzed. CONCLUSIONS: In pT2(+) patients the prognosis is significantly worse in multifocal involvement. In addition two groups of patients can be clearly distinguished from the BPFS point view according to their influential variables. The data suggest that since the prognostic point view the second group is understaged while the first is overstaged.

Graft survival and delayed graft function with normothermic regional perfusion and rapid recovery after circulatory death in kidney transplantation: a propensity score matching study.

BACKGROUND: A shortage of kidney grafts has led to the implementation of various strategies, including donations after circulatory death. The in situ normothermic regional perfusion technique has been introduced to improve graft quality by reducing warm ischemia times. However, there is limited evidence available on its mid- and long-term outcomes. Therefore, this study aimed to compare the incidence of delayed graft function, graft function, and survival at three years among three groups: brain death donors, rapid recovery, and normothermic regional perfusion. METHODS: A retrospective analysis of a cohort of kidney transplantations was conducted at a single referral center between January 1, 2015, and December 31, 2019. Univariate and multivariate regression models and propensity score matching analysis were performed to compare recipient-related, transplantation procedure-related, donor-related, and kidney function variables. RESULTS: A total of 327 patients were included, with 256 kidneys from brain death donors, 52 kidneys from rapid recovery, and 19 patients from normothermic regional perfusion. After propensity score matching, univariate and multivariate analyses showed a higher incidence of delayed graft function in the rapid recovery group compared to the others (OR: 2.39 CI95%: 1.19, 4.77) with a longer hospital stay (median 11, 15 and 10 days, respectively). However, no differences in 1- and 3-year graft function and survival were found. CONCLUSIONS: Normothermic regional perfusion offers advantages over rapid recovery, with a reduced incidence of delayed graft function and a shorter hospital stay. However, no differences in mid-term graft function and survival were found.

Effect of Heat Stress on Lactating and Non-Lactating Blackbelly Ewes under Tropical Conditions during Summer.

Two groups of ewes (10 lactating and 10 non-lactating) were used to evaluate the effect of heat stress during summer under tropical conditions. In this study, a temperature and humidity index (THI) was found that ranged between 65 and 79 (morning and afternoon). Likewise, a heat tolerance coefficient (HTC) of 6 units was observed. The highest breathing frequency (BF; 115.46 ± 35.25 breaths per minute (bpm)) and rectal temperature (RT; 38.95 ± 0.51 °C) were found during the afternoon in the group of lactating ewes. The means were compared by group, time of the day, and interaction, and only significant differences were found between groups for RT and udder temperature (p < 0.001). In the case of time of day, all parameters were higher during the afternoon, regardless of the group of ewes (p < 0.001). Likewise, an interaction was found in the parameters RT, right paralumbar fossa (RPF), rump, leg, and udder (p < 0.001). In conclusion, Blackbelly ewes lactating during the summer in the tropics have higher skin temperatures, and also raise BF and RT to tolerate HS in tropical climates.

Relative stability of human centrins and its relationship to calcium binding.

Centrins are calcium binding proteins that belong to the EF-hand superfamily with diverse biological functions. Herein we present the first systematic study that establishes the relative stability of related centrins via complementary biophysical techniques. Our results define the stepwise molecular behavior of human centrins by two-dimensional infrared (2D IR) correlation spectroscopy, the change in heat capacity and enthalpy of denaturation by differential scanning calorimetry, and the relative stability of the helical regions of centrins by circular dichroism. More importantly, 2D IR correlation spectroscopy provides unique information about the similarities and differences in dynamics between these related proteins. The thermally induced molecular behavior of human centrins can be used to predict biological target interactions that have a relative dependence on calcium affinity. This information is essential for understanding why certain isoforms may be used to rescue a phenotype and therefore also for explaining the different functions these proteins may have in vivo. Furthermore, this comparative approach can be applied to the study of recombinant therapeutic protein candidates for the treatment of disease states.