Pharmaceutical pollution of the world's rivers.

Environmental exposure to active pharmaceutical ingredients (APIs) can have negative effects on the health of ecosystems and humans. While numerous studies have monitored APIs in rivers, these employ different analytical methods, measure different APIs, and have ignored many of the countries of the world. This makes it difficult to quantify the scale of the problem from a global perspective. Furthermore, comparison of the existing data, generated for different studies/regions/continents, is challenging due to the vast differences between the analytical methodologies employed. Here, we present a global-scale study of API pollution in 258 of the world's rivers, representing the environmental influence of 471.4 million people across 137 geographic regions. Samples were obtained from 1,052 locations in 104 countries (representing all continents and 36 countries not previously studied for API contamination) and analyzed for 61 APIs. Highest cumulative API concentrations were observed in sub-Saharan Africa, south Asia, and South America. The most contaminated sites were in low- to middle-income countries and were associated with areas with poor wastewater and waste management infrastructure and pharmaceutical manufacturing. The most frequently detected APIs were carbamazepine, metformin, and caffeine (a compound also arising from lifestyle use), which were detected at over half of the sites monitored. Concentrations of at least one API at 25.7% of the sampling sites were greater than concentrations considered safe for aquatic organisms, or which are of concern in terms of selection for antimicrobial resistance. Therefore, pharmaceutical pollution poses a global threat to environmental and human health, as well as to delivery of the United Nations Sustainable Development Goals.

Accelerated Corneal Crosslinking to Arrest Progression of Corneal Ectasia: A Prospective Multicenter Study.

OBJECTIVES: To report the results of epithelium-off accelerated corneal collagen crosslinking (accelerated corneal crosslinking [ACXL]) in patients with progressive keratoconus. METHODS: This prospective, nonrandomized, noncomparative, interventional, multicenter clinical study included all patients who underwent ACXL, either continuous (c-ACXL; 9 mW/cm 2 , 10', 5.4 J/cm 2 ) or pulsed (p-ACXL; 2″ON/1″OFF, 30 mW/cm 2 , 4.5', 5.4 J/cm 2 ) between January 2014 and May 2017. Best-corrected visual acuity, sphere, cylinder, spherical equivalent, and topographical keratometry data were collected preoperatively and at 1, 3, 6, 12, 18, and 24 months postoperatively. RESULTS: Ninety-six eyes of 78 patients were included. The mean age was 20.8±4.4 years (14-33) for c-ACXL and 26.7±7.7 years (12-37) for p-ACXL. The mean best-corrected visual acuity was 0.4±0.4 for c-ACXL and 0.01±0.1 for p-ACXL preoperatively, and 0.3±0.3 ( P =0.0014) and -0.01±0.1 ( P =0.1554), respectively, at the last follow-up. The subjective sphere and spherical equivalent did not show statistically significant differences between the time points ( P >0.05). The subjective cylinder showed significant differences ( P =0.0013 for c-ACXL; P =0.0358 for p-ACXL). Keratometric values (K steep , K flat , and SimK) remained stable, with no statistically significant differences ( P >0.05). No major complications were noted. CONCLUSIONS: Both c-ACXL and p-ACXL are equally safe and effective ACXL protocols in stabilizing the progression of keratoconus and can be considered alternatives to the conventional Dresden protocol.

Clinical outcomes of traumatic globe rupture in corneal graft patients.

PURPOSE: This study reports the mechanisms, complications and graft survival following sight-threatening traumatic globe rupture in patients having previously undergone corneal transplantation in the same eye. METHODS: A retrospective, observational, single-center consecutive cohort study at the Royal Victoria Infirmary, Newcastle upon Tyne, UK over a 20-year period. Medical records and Newcastle Corneal Transplantation Service electronic database (eNCTS) review was undertaken of all consecutive patients who underwent corneal transplantation with a history of traumatic globe rupture. Main outcome measures include mechanism of injury, final best-corrected distance visual acuity (BCDVA), graft survival and complications. RESULTS: A total of 921 keratoplasties were undertaken between 1997 and 2017 with 24 (3.0%) patients identified with a history of traumatic globe rupture. A bimodal relationship of age and mechanism of trauma was observed. The mean age (SD) of individuals reporting cause as a fall was 71.5 (14.8) years, and 45.3 (20.8) years (P < 0.05) amongst individuals reporting accidental trauma or assault. The pre- and post-trauma mean (SD) LogMAR BCDVA was 0.6 (0.9) and 1.7 (1.0), respectively (P = 0.001). The overall graft-failure rate was 60.9% (11 grafts) during a mean (SD) follow-up period of 3.5 (4.1) years. Globe rupture with lens damage was associated with poorer final BCDVA (P < 0.05). CONCLUSIONS: This study represents the first published series from England for this type of patient cohort. Overall visual outcomes were poor with a bimodal relationship of age and mechanism of trauma. Worse prognostic factors included lens and posterior segment complications. Re-grafting in these select group of patients may prove valuable.

First report of Lasiodiplodia brasiliensis causing root rot in melon plants in northeastern Brazil.

Melon (Cucumis melo L.) is the second most exported fruit in Brazil with an annual production of 27.5 million tons (FAO 2023). From September 2019 through February 2020, 50-day-old melon plants started showing root rot symptoms (dark-brow necrotic zones in their roots that extended to the collar zone) in northeastern Brazil, 30% of the plants in the fields were affected by the disease. The fields are in clay soil where melon, in monoculture, is produced all year long with three cycles of the culture per year. A total of 132 samples from "Yellow" and "Cantaloupe" cultivars were collected from four melon fields (4°59'45.3"S, 37°33'39.7"W; 4°57'10.2"S, 37°31'37.1"W; 5°38'17.9"S, 37°56'27.7"W; and 5°00'25.5"S, 37°23'55.3"W). Small pieces of diseased tissues were surface disinfested in 70% ethanol for 30 sec, in 2% sodium hypochlorite for 1 min, washed in sterilized distilled water, plated on a PDA Petri dishes with tetracycline (0.05g/L), and incubated for seven days at 28 ± 2 ºC. Nine representative isolates were selected for downstream analysis. Colonies were white and later became dark gray, pycnidia and conidia were produced after 30 days ofncubation at 25°C under near-UV light in water-agar medium. Conidia were hyaline when immature and dark brown when mature, ranging from cylindrical subovoid to ellipsoidal and septate to non-septate, and with an average size of 12.54 to 21.97 µm. The colonies were morphologically identified as Lasiodiplodia sp. (Phillips et al. 2013). Total DNA from the isolates was extracted and the ITS, TUB, and TEF-1α genes (Jayawardena et al. 2019) were partially amplified by PCR, Sanger sequenced, and deposited in Genbank: ITS (OM102511 to OM102520), TUB (OR062087 to OR062094 and OR062095), and TEF-1α (OP536826 to OP536835). Blastn analysis of the partial sequences ITS (519bp), TUB (388bp), and TEF-1α (315bp) showed 100% nucleotide similarity of the isolates with sequences of L. brasiliensis and L. theobromae from the GenBank. A phylogenetic tree was constructed using the Maximum Parsimony Analysis method. All nine isolates were grouped into the L. brasiliensis clade with 71% bootstrap support, confirming the isolates's identity. Pathogenicity assays were conducted in a greenhouse using the wooden toothpick inoculation method (Nogueira et al. 2019). "Goldex" Yellow melon seedlings were used in a completely randomized experimental design, with 10 treatments (9 isolates + Mock) and six replicates, with one plant per pot. Plants were inoculated 15 days after sowing, and disease severity was evaluated 50 days after inoculation. All nine isolates caused symptoms in the assessed melon plants. The fungus was reisolated from the lesions and looked morphologically identical to the inoculated fungus, fulfilling Koch's postulates. The pathogenicity test was repeated and yielded similar results. All samples in this study were provided by melon growers who were concerned about the high incidence of root rot disease in their plantations. More research needs to be conducted to determine the epidemiology and the extension of the economic impact caused by this pathogen to melons to develop strategies for disease control to properly assist the growers's concerns. This pathogen has been reported to cause disease in other crops in Brazil, e.g., watermelon (Alves et al. 2023) and apples (Martins et al. 2018). However, to the best of our knowledge, this is the first report of L. brasiliensis causing root rot in melons in Brazil.

Aggressivity of Different Fusarium Species Causing Fruit Rot in Melons in Brazil.

Brazil is one of the largest melon (Cucumis melo) producers in the world and most of the production is exported to international markets. Currently, over 15% of Brazilian melon shipments are lost during export transportation due to Fusarium fruit rot, which is jeopardizing the livelihood of Brazilian melon producers. We focused on understanding the aggressivity of five species of Fusarium causing fruit rot on the main types of melon produced in Brazil. We also investigated the correlation between pathogenicity and fruit quality. Experiments were performed under a completely randomized experimental design, in a 5 × 8 factorial scheme, using two methods for inoculation: deposition of discs of culture media containing fungal structures and deposition of spore suspensions in needle-punctured lesions. The fungal species used were Fusarium falciforme, F. sulawesiense, F. pernambucanum, F. kalimantanense, and Fusarium sp. Fruits of two hybrids from four types of melons, canary (Goldex and Gold Mine), piel de sapo (Grand Prix and Flecha Verde), galia (McLaren and DRG3228), and cantaloupe (SV1044MF and Bonsai), were used. Disease severity was assessed by measuring the lesions, disease severity index, fruit firmness, and degrees Brix of fruits. The five Fusarium species caused rot in the fruits of all melon hybrids studied and the aggressivity of those fungal species varied with the type and hybrid. Fruits of the hybrids McLaren and Bonsai presented the largest lesions among all melon hybrids, and hybrids of canary type (Gold Mine and Goldex) were the most tolerant to rot caused by the Fusarium species investigated. Furthermore, the greater the severity of Fusarium fruit rot, the lower the pulp firmness of the fruits, but degrees Brix did not correlate with the onset of the disease.

Physical inactivity level and lipid profile in traditional communities in the Legal Amazon: a cross-sectional study : Physical inactivity level in the Legal Amazon.

BACKGROUND: The practice of regular physical activity can alter the lipid profile in populations according to diverse demographic characteristics. OBJECTIVE: To evaluate the association of physical inactivity with the lipid profile among vulnerable populations. METHODS: A cross-sectional study was conducted among 349 vulnerable individuals from Tocantins state, northern Brazil. The International Physical Activity Questionnaire 7 Day short form was used to measure self-reported physical activity levels. Venous blood samples were drawn to evaluate lipid profile. Logistic regression adjusted by the socioeconomic variables was used to analyse the effects of physical inactivity on the lipidic profile. The level of significance was 5% and Stata® (StataCorp, LC) version 11 was used. RESULTS: We observe an inverse relationship between physical inactivity and HDL-C-that is, those who were sedentary or below the WHO Recommendations for physical activity were at 2.6 greater odds (IC95% 1.21, 5.67; p = 0.015) of having a lower HDL compared with those meeting or exceeding WHO physical activity recommendations. CONCLUSION: On the vulnerable populations studied, the insufficiently active or sedentary individuals (called the physical inactivity individuals) have more risk of the altered HDL-C.

Effect of isolation method on human corneal stromal cell behaviour.

Current research on healthy corneal stromal cells will typically use primary cells as they are the most representative of in vivo behaviour. Primary cells are normally isolated from the limbus of discarded donor peripheral corneal tissue left over from transplantation (due to its relative abundance). Therefore, the central part of the cornea is less used in research as this tissue is usually used for transplantation. In some cases, although rare, the whole cornea, can become available for research. It is important to keep in mind that these corneas often have longer storage time, but the use of the central tissue for research is even more interesting, as knowing what cells are being transplanted into recipients would be highly relevant. To this end, stromal cells were extracted from both the limbus and central button of healthy corneas donated for research. This allowed for important comparison between central and limbal cells in culture. Of interest here was the extraction method of stromal cells from the donor tissue. The two most common methods of extraction are enzyme digestion and explant migration. However, no work has been done to understand how each method relatively affects the extracted cells. The extraction method and location from which stromal cells are harvested seems to have a significant effect on the cell adherence, survival, and gene expression of the stromal cells in culture. Enzyme digested cells showed that limbal and central cells had different gene expressions prior to culture, with gene such as ALDH3A1 being much more expressed in limbal cells. Enzyme digesting the limbal ring seems to yield the hardiest populations of stromal cells, a desirable trait in the culture of primary cells.

Relationship between income inequality, socioeconomic development, vulnerability index, and maternal mortality in Brazil, 2017.

BACKGROUND: Public health recognizes that health conditions depend on factors related to the development patterns income distribution, degree of poverty, working conditions, among other social determinants. The objective of this study was to analyze the association of maternal mortality with the Human Development Index (HDI), Gini Index, Income per capita, and the Social Vulnerability. METHOD: The study analyzed the relationship between MMR and socioeconomic indicators in the 26 federative units and the Federal District of Brazil, in 2017. The socioeconomic indicators used in the study were: HDI, Gini Index, Income per capita, and SVI. Crude and adjusted linear regression were performed between maternal mortality and socioeconomic indicators. RESULTS: When analyzing which socioeconomic determinants that are related to maternal mortality ratio rates, a higher per capita income positive effect was observed for lower MMR (ß = - 150.8; CI 95% -289.9 to - 11.7; r2 = 0.17; p = 0.035), as well as a trend of higher MMR in relation to the SVI (ß = 97.7; CI 95% -12.2 to 207.6; r2 = 0.12; p = 0.079). In model found by the stepwise forward selections, only the per capita income was um index related to less RMM (ß = - 0.02; CI 95% -0.05 to - 0.002; r2 = 0.15; p = 0.028). CONCLUSION: The findings showed that the per capita income has a negative association MMR in the different states of Brazil, but seems canceled because of the other socioeconomic determinants related to the poor live conditions.

Brazilian National Policy of Comprehensive Women's Health Care and mortality during climacteric period: has anything changed?

BACKGROUND: The National Policy for Integral Attention to Women's Health Care (PNAISM) was implemented in 2004, with monitoring of potential benefits. One of the life cycles of women contemplated in this health policy was the importance of health care during the climacteric. Prevention and health promotion are actions carried out by the Brazil National Health System and enshrined in health Brazilian policies for women. Thus, our purpose was to identify climacteric women's main causes of death as well as the mortality trends of such causes, especially after implementation of PNAISM. METHODS: An ecological study was conducted from 2018 to 2020. Data were retrieved from the Brazilian Health Department by accessing the mortality information system of the National Health Information, divided into periods 1996-2004 and 2005-2016 the latter to correspond with the implementation of the National Policy. The death records of Brazilian women aged 40 to 64 years who had a designated cause of death were retrieved. Trends and differences between periods were evaluated using linear regression. The significance level was set at 5%. RESULTS: The main causes of death in women from 1996 to 2016 were circulatory system diseases (22.47%, 697,636 deaths), neoplasms (19.69%, 611,495 deaths), respiratory system diseases (5.5%, 170,716 deaths), endocrine, nutritional, and metabolic disorders (5.27%, 163,602 deaths), and digestive system diseases (3.74%, 116.280 deaths). Analyzing the changes in the major causes of death of climacteric women after implementation of the PNAISM we observed that mortality from circulatory system diseases and endocrine and nutritional diseases were significantly declined in post-PNAISM period: (ß = - 3.63; 95% CI - 4.54 to - 2.73 r2 = 0.87; p < 0.001; ß = - 0.51; 95% CI, - 0.71 to - 0.31; R2 = 0.73; p < 0.001, respectively). No changes were observed in mortality from neoplasms and respiratory system diseases in post-PNAISM period (p = 0,765; p = 0,233, respectively). CONCLUSIONS: After implementation of the PNAISM, we observed a downward trend in rates of mortality from diseases of the circulatory and digestive systems and from endocrine, nutritional, and metabolic diseases but stability in the rates of death from neoplasm and respiratory system diseases.

Impact of biostimulant and saline water on cape gooseberry (Physalis peruviana L.) in Brazil.

Production of Physalis peruviana L. has gained prominence in Northeastern Brazil. However, salinity limits the crop development in the Brazilian semiarid. Thus, this research aimed to evaluate the application of Acadian® biostimulant as mitigant of the deleterious effects of salinity on growth and gas exchange of P. peruviana plants. The experiment was combining different electrical conductivity of irrigation water (0.50, 1.23, 3.00, 4.44, and 5.50 dS m-1) and biostimulant doses (0.00, 1.45, 5.00, 8.55, and 10.00 mL L-1). The main variables evaluated were plant height, stem diameter, number of leaves, root length, leaf area, specific leaf area, leaf area ratio, absolute and relative growth rate for plant height, and gas exchange. Experimental results showed that an increase in electrical conductivity of irrigation water had negatively affected the growth components and gas exchange in P. peruviana. Also, the application of seaweed-based biostimulant improves the photosynthetic capacity (43.3%), reduces transpiration rate (26.5%) and water loss by this process, further it attenuated the deleterious effects of salinity on specific leaf area, leaf area ratio, and stomatal conductance. To further elucidate the effectiveness of biostimulant application as a mitigant of salt stress, research aimed at the biochemical and enzyme activities of the plant's antioxidant system should be conducted to better understand this process.

The Role of Nerve Growth Factor in Maintaining Proliferative Capacity, Colony-Forming Efficiency, and the Limbal Stem Cell Phenotype.

Nerve growth factor (NGF) has demonstrated great benefit in the treatment of neurotrophic corneal ulcers. There is evidence for multiple modes of action in promoting corneal healing, but only indirect evidence exists for NGF's effects on limbal stem cells (LSCs). Understanding the role of NGF in LSC biology will improve our understanding of paracrine regulation of the limbal niche and the design of stem cell-based therapies for conditions such as LSC deficiency. In this article, we studied the regulation of NGF signaling components during LSC differentiation and the role of NGF in LSC proliferation and maintenance of the stem cell phenotype. LSC differentiation was induced by prolonged (40 day) culture which resulted in a significant increase in cell size, decrease in colony-forming efficiency and expression of putative LSC markers. A protein microarray measuring expression of 248 signaling proteins indicated the low affinity NGF receptor p75NTR to be the most downregulated protein upon differentiation. Further confirmation by Western blotting and real-time quantitative polymerase chain reaction indicated that NGF and p75NTR are expressed in early LSC cultures and downregulated upon differentiation. LSC cultures grown in the presence of anti-NGF antibody showed decreased colony-forming efficiency, DNA replication and expression of putative LSC markers ABCG2 and C/EBPδ. Supplementation of LSC culture medium with NGF extended the life span of LSC cultures in vitro and increased the expression of putative LSC markers ΔNp63α and ABCG2. Taken together, our data indicate that NGF signaling is a key promoter of LSC proliferation, colony-forming efficiency, and a maintainer of the LSC phenotype. Stem Cells 2019;37:139-149.

Correction to: Regional disparities in the flow of access to breast cancer hospitalizations in Brazil in 2004 and 2014.

An amendment to this paper has been published and can be accessed via the original article.

Regional disparities in the flow of access to breast cancer hospitalizations in Brazil in 2004 and 2014.

BACKGROUND: Access to the diagnosis and treatment of breast cancer in Brazil is marked by immense inequalities in the provision of specialized assistance, which leads patients to seek treatment outside the place of residence. To evaluate the variations between 2004 and 2014 in the distribution of flow between place of residence and care, and the average distance traveled for treatment of breast cancer in the administrative regions and federal states of Brazil. METHOD: Analysis of secondary data from the years 2004 and 2014, extracted from the Department of Informatics of the Unified Health System through the Hospital Information System. Data from Hospitalization Release Authorizations were collected, and the maps were created with TabWin 3.6 software. Descriptive analysis was performed on Stata® (StataCorp, LC) 11.0. RESULTS: In the total flow, it was observed that there was a decrease in referrals between 2004 and 2014 in most regions. In 2004 the main direction of flow was in the Midwest and Southeast regions. In 2014, however, the intensity of these admissions was centralized in the Southeast region. In relation to the average distance traveled, the North, Northeast, and Midwest regions had the highest values of displacement. Of the 27 federative units, 17 presented an increase in average distance between these periods. CONCLUSION: Despite the improvement in the hospitalization of residents, in most regions and federal units, Brazilians still travel great distances when they require treatment for breast cancer.

Do assisted reproduction outcomes differ according to aetiology of obstructive azoospermia?

Azoospermia is defined as absence of spermatozoa and may be secondary to blocked seminal ducts, known as obstructive azoospermia. Semen quality may be impaired due to factors such as sperm cell DNA fragmentation and presence of antisperm antibodies. The objective of this article was to investigate potential differences in outcomes of in vitro fertilisation and intracytoplasmic sperm injection between groups with different obstruction aetiology, as well as between the use of different techniques and sperm cells of different origins. Retrospective, multi-centre analysis of 621 first cycles was carried out between 2008 and 2015: Group I, congenital obstruction, 45 patients and Group 2, vasectomy, 576 patients. Sperm cell retrieval was achieved in all cases. Results were similar for Group I and II fertilisation rates, 70% versus 66.85% (p = .786); pregnancy rates, 42.5% versus 41.46% (p = .896); and live birth rates, 29.73% versus 17.69% (p = .071). According to sperm cell origin (579 epididymal vs. 42 testicular), pregnancy rates, 41.47% versus 43.9% (p = .760); and live birth rates, 18.3% versus 27.78% (p = .163) had no difference. Fertilisation, pregnancy and live birth rates did not differ according to obstruction aetiology. Outcomes did not differ between groups according to sperm cell origin.

Hypothermically Stored Adipose-Derived Mesenchymal Stromal Cell Alginate Bandages Facilitate Use of Paracrine Molecules for Corneal Wound Healing.

Adipose-derived mesenchymal stromal cells (Ad-MSCs) may alleviate corneal injury through the secretion of therapeutic factors delivered at the injury site. We aimed to investigate the therapeutic factors secreted from hypothermically stored, alginate-encapsulated Ad-MSCs' bandages in in vitro and in vivo corneal wounds. Ad-MSCs were encapsulated in 1.2% w/v alginate gels to form bandages and stored at 15 °C for 72 h before assessing cell viability and co-culture with corneal scratch wounds. Genes of interest, including HGF, TSG-6, and IGF were identified by qPCR and a human cytokine array kit used to profile the therapeutic factors secreted. In vivo, bandages were applied to adult male mice corneas following epithelial debridement. Bandages were shown to maintain Ad-MSCs viability during storage and able to indirectly improve corneal wound healing in vivo. Soluble protein concentration and paracrine factors such as TSG-6, HGF, IL-8, and MCP-1 release were greatest following hypothermic storage. In vivo, Ad-MSCs bandages-treated groups reduced immune cell infiltration when compared to untreated groups. In conclusion, bandages were shown to maintain Ad-MSCs ability to produce a cocktail of key therapeutic factors following storage and that these soluble factors can improve in vitro and in vivo corneal wound healing.

Outcomes of Penetrating Keratoplasty Following Autologous Cultivated Limbal Epithelial Stem Cell Transplantation.

The purpose of this study is to investigate the outcomes of penetrating keratoplasty (PKP) following autologous cultivated limbal epithelial stem cell transplantation (CLET). A prospective, single center, interventional cohort study investigating patients with unilateral total limbal stem cell deficiency (LSCD) treated with CLET who underwent PKP. Patients with confirmed corneal re-epithelialization > 6 months post-CLET, and with best-corrected visual acuity (BCVA) <0.3 logMAR were offered PKP. CLET survival assessed by slit lamp, corneal impression cytology (CIC), and in vivo confocal microscopy. Confirmation of corneal re-epithelialization by histological and immunocytochemical (ICC) examination of trephined corneal buttons. Mean change in best-corrected visual acuity (logMAR) following PKP and PKP survival at 12 months were calculated. Twenty patients underwent PKP. Mean time of PKP was 19 months (range 11-41 months, SD 7.26) post-CLET. Median follow-up time post-PKP was 15 months (range 1-32, SD 10.2). CIC and ICC of all corneas confirmed corneal re-epithelialization before PKP. Mean pre-PKP BCVA was 1.46 (range 0.3-2.7, SD 0.94) improving to a mean post-PKP BCVA of 0.74 (range 0-2.7, SD 0.87); mean improvement in BCVA post-PKP of 36 letters (95% CI 15.0-57.1, p = .002). Kaplan-Meier mean graft survival was 90.9% (95% CI 50.8-98.7) at 12 months. We recommend a two-stage approach with CLET followed by PKP >12 months later. Patients experienced a significant improvement in BCVA following PKP. PKP did not have a detrimental effect on CLET survival. PKP survival post-CLET is better than that reported for high risk PKP. Stem Cells 2018;36:925-931.

CD200 Expression Marks a Population of Quiescent Limbal Epithelial Stem Cells with Holoclone Forming Ability.

One of the main challenges in limbal stem cell (LSC) biology and transplantation is the lack of definitive cell surface markers which can be used to identify and enrich viable LSCs. In this study, expression of 361 cell surface proteins was assessed in ex vivo expanded limbal epithelial cells. One marker, CD200 was selected for further characterization based on expression in a small subset of limbal epithelial cells (2.25% ± 0.69%) and reduced expression through consecutive passaging and calcium induced differentiation. CD200 was localized to a small population of cells at the basal layer of the human and mouse limbal epithelium. CD200+ cells were slow cycling and contained the majority of side population (SP) and all the holoclone forming progenitors. CD200+ cells displayed higher expression of LSCs markers including PAX6, WNT7A, CDH3, CK14, CK15, and ABCB5 and lower expression of Ki67 when compared to CD200- . Downregulation of CD200 abrogated the ability of limbal epithelial cells to form holoclones, suggesting an important function for CD200 in the maintenance and/or self-renewal of LSCs. A second marker, CD109, which was expressed in 56.29% ± 13.96% of limbal epithelial cells, was also found to co-localize with ΔNp63 in both human and mouse cornea, albeit more abundantly than CD200. CD109 expression decreased slowly through calcium induced cell differentiation and CD109+ cells were characterized by higher expression of Ki67, when compared to CD109- subpopulation. Together our data suggest that CD200 expression marks a quiescent population of LSCs with holoclone forming potential, while CD109 expression is associated with a proliferative progenitor phenotype. Stem Cells 2018;36:1723-1735.

Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial-Like Cells.

Cornea is a clear outermost layer of the eye which enables transmission of light onto the retina. The transparent corneal epithelium is regenerated by limbal stem cells (LSCs), whose loss/dysfunction results in LSCs deficiency (LSCD). Ex vivo expansion of autologous LSCs obtained from patient's healthy eye followed by transplantation onto the LSCs damaged/deficient eye, has provided a successful treatment for unilateral LSCD. However, this is not applicable to patient with total bilateral LSCD, where LSCs are lost/damaged from both eyes. We investigated the potential of human induced pluripotent stem cell (hiPSC) to differentiate into corneal epithelial-like cells as a source of autologous stem cell treatment for patients with total bilateral LSCD. Our study showed that combined addition of bone morphogenetic protein 4 (BMP4), all trans-retinoic acid and epidermal growth factor for the first 9 days of differentiation followed by cell-replating on collagen-IV-coated surfaces with a corneal-specific-epithelial cell media for an additional 11 days, resulted in step wise differentiation of human embryonic stem cells (hESC) to corneal epithelial progenitors and mature corneal epithelial-like cells. We observed differences in the ability of hiPSC lines to undergo differentiation to corneal epithelial-like cells which were dependent on the level of endogenous BMP signaling and could be restored via the activation of this signaling pathway by a specific transforming growth factor ß inhibitor (SB431542). Together our data reveal a differential ability of hiPSC lines to generate corneal epithelial cells which is underlined by the activity of endogenous BMP signaling pathway. Stem Cells 2018;36:337-348.

Time trend of breast cancer mortality in BRAZILIAN men: 10-year data analysis from 2005 to 2015.

BACKGROUND: Male Breast Cancer (MBC) is rare, which makes its understanding and treatment be extrapolated from what is known about the occurrence in women, with few epidemiological studies, with few epidemiological studies. Therefore, the aim of the present paper was to study breast cancer mortality in adult males in Brazil and its administrative regions between 2005 and 2015. METHODS: Ecological study with data on MBC mortality in adults between 2005 and 2015. Data were obtained from the Mortality Information System of the Department of Informatics of SUS (the Unified Health System of the country). Descriptive statistics were used for MBC mortality and linear regression to analyze the relationship between mortality and the country's administrative regions. Percentage Change (PC) and Annual Percentage Change (APC) were the trend measures used for MBC mortality for the period. RESULTS: Between 2005 and 2015, there were 1521 deaths due to MBC in adults in Brazil. Regarding mortality by region, there was great oscillation in the rates of the country as a whole (PC = 113,87; ß = 0,009 (IC95% 0,000 - 0,018); r2 = 0,381; P = 0,043). The highest increase in MBC mortality occurred in patients aged 80 years or older (PC = 161,04; ß = 0,201 (IC95% 0,640 - 0,339); r2 = 0,550; P = 0,009) and there was significant increase in deaths for the 50-54-year age group (PC = 224,01; ß = 0,135 (CI95% 0,052; 0,218); r2 = 0,601; P = 0,005). CONCLUSION: Mortality in adults due to MBC increased in Brazil during the study period with the highest percentage increase occurring for individuals aged 80 years or older.

Association between vitamin D plasma concentrations and VDR gene variants and the risk of premature birth.

BACKGROUND: Premature birth is the main cause of mortality in children under 1 year, and vitamin D deficiency during gestation is associated with prematurity. The effects of vitamin D are mediated by its receptor, which is encoded by the VDR gene. VDR variants-such as single nucleotide variation (SNV)-are associated with increased risk of prematurity, but there are conflicting results. We evaluated serum vitamin D concentrations and the frequency of TaqI/A > G, BsmI/C > T, ApaI/C > A, and FokI/A > T VDR variants in mothers and preterm (PTN) and full-term (FTN) newborns. METHODS: We conducted a case-control study comprising 40 pairs of mothers and their PTNs (gestational age < 32 weeks and/or weight < 1500 g), and 92 pairs of mothers and FTNs as controls. Genotyping was performed by real-time PCR, and plasma vitamin D concentrations were measured by electrochemiluminescence. RESULTS: Vitamin D levels were significantly lower in PTN mothers. Genotypes TaqI/GG and BsmI/TT, and haplotypes AAG (TaqI/A-ApaI/A-FokI/G) and GCA (TaqI/G-ApaI/C-FokI/A) were significantly more frequent in PTN mothers, and genotypes TaqI/AG, ApaI/AA, and FokI/AG resulted in significantly lower vitamin D levels. Genotypes BsmI/TT and ApaI/AA were associated with vitamin D deficiency and 2.36 and 7.99 times greater likelihood of PTB, respectively. Vitamin D levels were also lower in PTNs, although it was not statistically significant. Genotypes BsmI/TT, ApaI/AA, and FokI/GG, and haplotype GAG (TaqI/G-ApaI/A-FokI/G) were significantly more frequent in PTNs. Those with FokI/GG genotypes had significantly lower vitamin D levels. CONCLUSIONS: VDR variants contribute to variations in vitamin D concentrations and the increased risk of prematurity.