The need for culture in tegumentary leishmaniasis diagnosis in Bolivia: A comparative evaluation of four parasitological techniques using two sampling methods.

Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. In Bolivia, leishmaniasis occurs mainly in the cutaneous form (CL) followed by the mucosal or mucocutaneous form (ML or MCL), grouped as tegumentary leishmaniosis (TL), while cases of visceral leishmaniasis (VL) are rare. The cases of TL are routinely diagnosed by parasitological methods: Direct Parasitological Exam (DPE) and axenic culture, the latter being performed only by specialized laboratories. The aim of the present study was to optimize the parasitological diagnosis of TL in Bolivia, using two sampling methods. Samples from 117 patients with suspected TL, obtained by aspiration (n = 121) and scraping (n = 121) of the edge of the lesion were tested by: direct parasitological exam, culture in TSTB medium, and miniculture and microculture in Schneider's medium. A positive laboratory result by any of the four techniques evaluated using either of the two sampling methods was considered the gold standard. Of the 117 suspected patients included, TL was confirmed in 96 (82 %), corresponding 79 of the confirmed cases (82.3 %) to CL and 16 (16.7 %) to ML. Parasitological techniques specificity was 100 % and their analytical sensitivity was greater with scraping samples in TSTB culture (98 %). Scraping samples in TSTB and miniculture correlated well with the reference (Cohen's kappa coefficient=0.88) and showed good reliability (Cronbach's alpha coefficient ≥0.91). Microculture provided positive results earlier than the other culture methods (mean day 4.5). By day 14, 98 % of positive cultures had been detected. Scraping sampling and miniculture were associated with higher culture contamination (6 % and 17 %, respectively). Bacterial contamination predominated, regardless of the sampling and culture method, while filamentous fungi and mixed contamination were more frequently observed in cultures from scraping samples. In conclusion: (i) scraping samples proved more suitable for the diagnosis of TL as they increased analytical sensitivity, are less traumatic for the patient and are safer for laboratory personnel than aspirates; (ii) culture, mainly in TSBT medium, should be used for the diagnosis of TL due to its high sensitivity (doubling the number of cases diagnosed by DPE) and its low cost compared to other culture media.

Usefulness of Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in the Characterization of Leishmania Strains Causing Tegumentary Leishmaniasis in Bolivia versus hsp70 Gene Sequencing.

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is a proteomic technique with proven efficiency in the identification of microorganisms, such as bacteria, fungi, and parasites. The present study aimed to evaluate the usefulness of MALDI-TOF MS for the characterization of Leishmania species circulating in Bolivia using hsp70 gene sequencing as a reference technique. 55 Leishmania strains that were isolated from patients with tegumentary leishmaniasis were analyzed. MALDI-TOF MS identified two species of the L. braziliensis complex (L. braziliensis, n = 26; L. braziliensis outlier, n = 18), one species of the L. guyanensis complex (L. guyanensis, n = 1), one species of the L. lainsoni complex (L. lainsoni, n = 2), and two species of the L. mexicana complex (L. amazonensis, n = 5; and L. garnhami, n = 3). All of the strains were correctly identified at the subgenus, genus, and complex level, but 10 of them (18%) were misidentified as other species within the same complex by the hsp70 gene sequencing, with 7 of these corresponding to possible hybrids. Thus, one L. braziliensis corresponded to L. peruviana, two L. braziliensis corresponded to L. braziliensis/L. peruviana possible hybrids, two L. amazonensis corresponded to L. mexicana, and three L. garnhami and two L. amazonensis corresponded to L. mexicana/L. amazonensis possible hybrids. Accordingly, MALDI-TOF MS could be used as an alternative to molecular techniques for the identification of Leishmania spp., as it is low cost, simple to apply, and able to quickly produce results. In Bolivia, its application would allow for the improvement of the management of patient follow-ups, the updating of the epidemiological data of the Leishmania species, and a contribution to the control of tegumentary leishmaniasis. IMPORTANCE The objective of the study was to evaluate the usefulness of MALDI-TOF MS for the characterization of Leishmania species circulating in Bolivia, in comparison with the sequencing of the hsp70 gene. In our study, all of the isolates could be identified, and no misidentifications were observed at the complex level. Although the equipment implies a high initial investment in our context, MALDI-TOF MS can be used in different areas of microbiology and significantly reduces the cost of testing. Once the parasite culture is obtained, the technique quickly yields information by accessing a free database that is available online. This would allow for the improvement of the management of patients and follow-ups, the updating of the epidemiological data of the species, and a contribution to the control of tegumentary leishmaniasis in Bolivia. Likewise, it can be used to determine a specific treatment to be given, according to the causal species of Leishmania, when there are protocols in this regard in the area.

Cutaneous leishmaniasis treatment and therapeutic outcomes in special populations: A collaborative retrospective study.

BACKGROUND: Treatment guidance for children and older adult patients affected by cutaneous leishmaniasis (CL) is unclear due to limited representation of these groups in clinical trials. METHODS: We conducted a collaborative retrospective study to describe the effectiveness and safety of antileishmanial treatments in children ≤ 10 and adults ≥ 60 years of age, treated between 2014 and 2018 in ten CL referral centers in Latin America. RESULTS: 2,037 clinical records were assessed for eligibility. Of them, the main reason for non-inclusion was lack of data on treatment follow-up and therapeutic response (182/242, 75% of children and 179/468, 38% of adults). Data on 1,325 eligible CL patients (736 children and 589 older adults) were analyzed. In both age groups, disease presentation was mild, with a median number of lesions of one (IQR: 1-2) and median lesion diameter of less than 3 cm. Less than 50% of the patients had data for two or more follow-up visits post-treatment (being only 28% in pediatric patients). Systemic antimonials were the most common monotherapy regimen in both age groups (590/736, 80.2% of children and 308/589, 52.3% of older adults) with overall cure rates of 54.6% (95% CI: 50.5-58.6%) and 68.2% (95% CI: 62.6-73.4%), respectively. Other treatments used include miltefosine, amphotericin B, intralesional antimonials, and pentamidine. Adverse reactions related to the main treatment were experienced in 11.9% (86/722) of children versus 38.4% (206/537) of older adults. Most adverse reactions were of mild intensity. CONCLUSION: Our findings support the need for greater availability and use of alternatives to systemic antimonials, particularly local therapies, and development of strategies to improve patient follow-up across the region, with special attention to pediatric populations.

Tegumentary leishmaniasis by Leishmania braziliensis complex in Cochabamba, Bolivia including the presence of L. braziliensis outlier: Tegumentary leishmaniasis in Cochabamba, Bolivia: Tegumentary leishmaniasis in Cochabamba, Bolivia.

Leishmaniasis is caused by protozoans of the Leishmania genus, which includes more than 20 species capable of infecting humans worldwide. In the Americas, the most widespread specie is L. braziliensis, present in 18 countries including Bolivia. The taxonomic position of the L. braziliensis complex has been a subject of controversy, complicated further by the recent identification of a particular subpopulation named L. braziliensis atypical or outlier. The aim of this study was to carry out a systematic analysis of the L. braziliensis complex in Bolivia and to describe the associated clinical characteristics. Forty-one strains were analyzed by sequencing an amplified 1245 bp fragment of the hsp70 gene, which allowed its identification as: 24 (59%) L. braziliensis, 16 (39%) L. braziliensis outlier, and one (2%) L. peruviana. In a dendrogram constructed, L. braziliensis and L. peruviana are grouped in the same cluster, whilst L. braziliensis outlier appears in a separate branch. Sequence alignment allowed the identification of five non-polymorphic nucleotide positions (288, 297, 642, 993, and 1213) that discriminate L. braziliensis and L. peruviana from L. braziliensis outlier. Moreover, nucleotide positions 51 and 561 enable L. peruviana to be discriminated from the other two taxa. A greater diversity was observed in L. braziliensis outlier than in L. braziliensis-L. peruviana. The 41 strains came from 32 patients with tegumentary leishmaniasis, among which 22 patients (69%) presented cutaneous lesions (11 caused by L. braziliensis and 11 by L. braziliensis outlier) and 10 patients (31%) mucocutaneous lesions (eight caused by L. braziliensis, one by L. braziliensis outlier, and one by L. peruviana). Nine patients (28%) simultaneously provided two isolates, each from a separate lesion, and in each case the same genotype was identified in both. Treatment failure was observed in six patients infected with L. braziliensis and one patient with L. peruviana.

Pt-Free Metal Nanocatalysts for the Oxygen Reduction Reaction Combining Experiment and Theory: An Overview.

The design and manufacture of highly efficient nanocatalysts for the oxygen reduction reaction (ORR) is key to achieve the massive use of proton exchange membrane fuel cells. Up to date, Pt nanocatalysts are widely used for the ORR, but they have various disadvantages such as high cost, limited activity and partial stability. Therefore, different strategies have been implemented to eliminate or reduce the use of Pt in the nanocatalysts for the ORR. Among these, Pt-free metal nanocatalysts have received considerable relevance due to their good catalytic activity and slightly lower cost with respect to Pt. Consequently, nowadays, there are outstanding advances in the design of novel Pt-free metal nanocatalysts for the ORR. In this direction, combining experimental findings and theoretical insights is a low-cost methodology-in terms of both computational cost and laboratory resources-for the design of Pt-free metal nanocatalysts for the ORR in acid media. Therefore, coupled experimental and theoretical investigations are revised and discussed in detail in this review article.

Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis.

This protocol identifies the immunity proteins of the bactericidal enzymes: colicin E3 and bacteriocin, produced by a pathogenic Escherichia coli strain using antibiotic induction, and identified by MALDI-TOF-TOF tandem mass spectrometry and top-down proteomic analysis with software developed in-house. The immunity protein of colicin E3 (Im3) and the immunity protein of bacteriocin (Im-Bac) were identified from prominent b- and/or y-type fragment ions generated by the polypeptide backbone cleavage (PBC) on the C-terminal side of aspartic acid, glutamic acid, and asparagine residues by the aspartic acid effect fragmentation mechanism. The software rapidly scans in silico protein sequences derived from the whole genome sequencing of the bacterial strain. The software also iteratively removes amino acid residues of a protein sequence in the event that the mature protein sequence is truncated. A single protein sequence possessed mass and fragment ions consistent with those detected for each immunity protein. The candidate sequence was then manually inspected to confirm that all detected fragment ions could be assigned. The N-terminal methionine of Im3 was post-translationally removed, whereas Im-Bac had the complete sequence. In addition, we found that only two or three non-complementary fragment ions formed by PBC are necessary to identify the correct protein sequence. Finally, a promoter (SOS box) was identified upstream of the antibacterial and immunity genes in a plasmid genome of the bacterial strain.

Clinical and immunological characteristics of tegumentary leishmaniasis cases in Bolivia.

BACKGROUND: Tegumentary leishmaniasis (TL) is a parasitic disease that can present a cutaneous or mucocutaneous clinical form (CL and MCL, respectively). The disease is caused by different Leishmania species and transmitted by phlebotomine sand flies. Bolivia has one of the highest incidences of the disease in South America and the diagnosis is done by parasitological techniques. Our aim was to describe the clinical and immunological characteristics of CL and MCL patients attending the leishmaniasis reference center in Cochabamba, Bolivia, in order to gain updated clinical and epidemiological information, to evaluate the diagnostic methods used and to identify biomarkers related to clinical disease and its evolution. METHODOLOGY/PRINCIPAL FINDINGS: The study was conducted from September 2014 to November 2015 and 135 patients with lesions compatible with CL or MCL were included. Epidemiological and clinical data were collected using a semi-structured questionnaire. Two parasitological diagnostic methods were used: Giemsa-stained smears and culture of lesion aspirates. Blood samples obtained from participants were used to measure the concentrations of different cytokines. 59.2% (80/135) were leishmaniasis confirmed cases (CL: 71.3%; MCL: 28.7%). Sixty percent of the confirmed cases were positive by smears and 90.6% were positive by culture. 53.8% were primo-infections. Eotaxin and monokine induced by IFN-γ presented higher serum concentrations in the MCL clinical presentation compared to CL cases and no-cases. None of the cytokines presented different concentrations between primo-infections and secondary infections due to treatment failure. CONCLUSIONS/SIGNIFICANCE: In Bolivia, parasitological diagnosis remains the reference standard in diagnosis of leishmaniasis because of its high specificity, whereas the sensitivity varies over a wide range leading to loss of cases. Until more accurate tools are implemented, all patients should be tested by both smears and culture of lesion aspirates to minimize the risk of false negatives. Our results showed higher concentrations of several cytokines in MCL compared to CL, but no differences were observed between CL and no-cases. In addition, none of the cytokines differed between primary and secondary infections. These results highlight the need of further research to identify biomarkers of susceptibility and disease progression, in addition to looking at the local cellular immune responses in the lesions.

Single-cell analysis of the developing human testis reveals somatic niche cell specification and fetal germline stem cell establishment.

Human testis development in prenatal life involves complex changes in germline and somatic cell identity. To better understand, we profiled and analyzed ∼32,500 single-cell transcriptomes of testicular cells from embryonic, fetal, and infant stages. Our data show that at 6-7 weeks postfertilization, as the testicular cords are established, the Sertoli and interstitial cells originate from a common heterogeneous progenitor pool, which then resolves into fetal Sertoli cells (expressing tube-forming genes) or interstitial cells (including Leydig-lineage cells expressing steroidogenesis genes). Almost 10 weeks later, beginning at 14-16 weeks postfertilization, the male primordial germ cells exit mitosis, downregulate pluripotent transcription factors, and transition into cells that strongly resemble the state 0 spermatogonia originally defined in the infant and adult testes. Therefore, we called these fetal spermatogonia "state f0." Overall, we reveal multiple insights into the coordinated and temporal development of the embryonic, fetal, and postnatal male germline together with the somatic niche.

Transformation of Nanostructures Cu2O to Cu3Se2 through Different Routes and the Effect on Photocatalytic Properties.

In this work, copper selenide (Cu3Se2 umangite phase) was synthesized by two routes, using a chemical reaction and the hydrothermal method to obtain CuSe-A and CuSe-H, respectively. The synthesis of Cu3Se2 consisted of a three-step process: in the first step, copper(I) oxide hexapods (Cu2O) were obtained as the copper reservoir; in the second step, selenium ions were obtained from the reduction of selenium powder; and in the third step involves mixing two precursors following the two synthesis routes mentioned before. Analysis of X-ray diffraction and X-ray photoelectron spectroscopy showed the formation of the Cu3Se2 phase by both synthesis routes. On the other hand, using the scanning electron microscopy (SEM) technique, it is observed that the Cu3Se2 sample (CuSe-A) is obtained by exchanging in solution with agitation and that the copper selenium phase grows only on the surface of the hexapods. Meanwhile, the hydrothermal route promotes a total conversion of copper(I) oxide hexapods to the copper selenide phase (CuSe-H). The resulting materials were tested as photocatalytic materials to remove methylene blue dye in water under sunlight irradiation. Cu3Se2 (CuSe-H) obtained by the hydrothermal route exhibited a higher efficiency of photodegradation of dye, reaching a removal percentage of 92% after 4 h under sunlight.

INTESTINAL AND BLOOD PARASITES IN SCARLET (ARA MACAO) AND GREAT GREEN (ARA AMBIGUA) MACAWS IN WILDLIFE REHABILITATION CENTERS IN COSTA RICA.

Costa Rica undertakes continuous efforts to recover the native population of macaw species through rehabilitation programs for breeding and releasing birds in protected areas. In the summer of 2018, a total of 107 scarlet (Ara macao) and 93 great green (Ara ambigua) macaws were sampled in four wildlife rehabilitation centers in Costa Rica. Fecal samples representing 200 individuals were analyzed for intestinal parasites, and 23 individuals were sampled for hemoparasites. Ascaridia and Capillaria were found in fecal samples. No hemoparasites were found. The distribution of percentage of infection was analyzed by location, species, and housing type. As part of a health screening prior to release, parasitological examination is recommended.

Mini-percutaneous nephrolithotomy versus retrograde intrarenal surgery for the treatment of 10-20 mm lower pole renal stones: a systematic review and meta-analysis.

PURPOSE: To compare the efficacy and safety of mini-percutaneous nephrolithotomy (mini-PCNL) versus retrograde intrarenal surgery (RIRS) for treatment of 10-20 mm lower pole renal stones. METHODS: A comprehensive literature search of PubMed, Scopus, the Cochrane Library, and Web of Science was conducted to identify all studies comparing mini-PCNL and RIRS for 10-20 mm lower pole renal stones before March 2019. Article selection proceeded according to the search strategy based on the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The meta-analysis was performed with the R program version 3.5.1. RESULTS: A total of five studies were included (two randomized controlled trials and three case-controlled trials) with a total of 587 patients included. The success rate was significantly higher in the mini-PCNL group (OR 1.67; 95% CI p = 0.05). Operative and fluoroscopy times were similar for both groups (MD 2.45; 95% CI p = 0.87 and MD 2.11; 95% CI p = 0.09, respectively). Concerning the hospital stay and overall complication rates, there were no differences between the two procedures (MD 41.94; 95% CI p = 0.18 and OR 1.76; 95% CI p = 0.11). CONCLUSION: Our analysis showed that both procedures are safe for treatment of 10-20 mm lower pole renal stones with similar complication rates, operative times, fluoroscopy times and length of hospital stay, but mini-PCNL was significantly superior in effectiveness with a higher success rate. Based on these results, mini-PCNL may be included in the guidelines as a safe and effective alternative treatment for 10-20 mm lower pole stones.

Generation of three human induced pluripotent stem cell sublines (MZT04D, MZT04J, MZT04C) for reproductive science research.

We generated three human induced pluripotent stem cell (hiPSC) sublines from human dermal fibroblasts (HDFs) (MZT04) generated from a skin biopsy donated from a previously fertile woman. The skin biopsy was broadly consented for generating hiPSC lines for biomedical research, including unique consent specifically for studying human fertility, infertility and germ cells. hiPSCs were reprogrammed using Sendai virus vectors and were subsequently positive for markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA-4. Pluripotency was further verified using teratomas and PluriTest. These sublines serve as controls for hiPSC research projects aimed at understanding the cell and molecular regulation of female fertility and infertility.

Evaluación hepática y renal en pacientes que recibieron conjuntamente zinc y tratamiento antituberculosis acortado / Hepatic and renal evaluation in patients that received jointly zinc and antituberculosis shortened treatment

La terapia de la tuberculosis con el esquema 2RHZE/4HE comprende la administración durante seis meses de rifampicina, isoniazida, pirazinamida y etambutol, de las cuales las tres primeras son potencialmente hepatotoxicas y excepcionalmente nefrotoxicas. La tuberculosis produce depleción de la concentración de zinc lo que incrementa la susceptibilidad a la cronicidad de la infección. OBJETIVO: evaluar el efecto de la administración conjunta de zinc y la terapia 2RHZE/4HE sobre la función hepática y renal en pacientes con tuberculosis pulmonar. MÉTODOS: estudio descriptivo de tipo caso control, doble ciego aleatorizado con 22 pacientes con tuberculosis pulmonar en terapia farmacológica y 22 controles sanos reclutados en los centros de salud Sebastián pagador y Alalay. Los pacientes fueron divididos aleatoriamente en dos grupos a los que se les administró zinc (45mg/día) o placebo durante tres meses. En todos los pacientes se tomó muestras de sangre antes y después de la intervención para medir pruebas de función renal y hepática. En los sujetos control la muestra de sangre se tomó al inicio del estudio para realizar las mismas determinaciones. RESULTADOS: no se encontró deferencias en la concentración de marcadores específicos de daño hepático o renal. CONCLUSIONES: la adición de un suplemento diario de 45mgr de zinc a la terapia 2RHZE/4HE no produjo daño renal ni hepático en las personas evaluadas.

Tuberculosis therapy with the 2RHZE / 4HE scheme comprises the administration for six months of rifampin, isoniazid, pyrazinamide and ethambutol, of which the first three are potentially hepatotoxic and exceptionally nephrotoxic. Tuberculosis produces depletion of the zinc concentration which increases the susceptibility to chronicity of the infection. OBJECTIVE: to evaluate the effect of co-administration of zinc and 2RHZE /4HE therapy on hepatic and renal function in patients with pulmonary tuberculosis. METHODS: a, descriptive case-control, randomized double-blind study. 22 patients with pulmonary tuberculosis receiving pharmacological therapy and 22 healthy controls recruited in the health centers Sebastián Pagador and Alalay. The patients were randomly divided into two groups. Who were given zinc (45mg / day) or placebo for three months. All patients were blood sampling before and after intervention to measure hepatic and renal functional tests. For the control subjects were blood sampling before the study to do the same test. RESULTS: no deference was found in the concentration of specific markers of hepatic or renal damage. CONCLUSIONS: the addition of a daily supplement of 45 mg of zinc to 2RHZE/4HE therapy did not cause hepatic, neither renal damage in the people evaluated.

Differentiation of primate primordial germ cell-like cells following transplantation into the adult gonadal niche.

A major challenge in stem cell differentiation is the availability of bioassays to prove cell types generated in vitro are equivalent to cells in vivo. In the mouse, differentiation of primordial germ cell-like cells (PGCLCs) from pluripotent cells was validated by transplantation, leading to the generation of spermatogenesis and to the birth of offspring. Here we report the use of xenotransplantation (monkey to mouse) and homologous transplantation (monkey to monkey) to validate our in vitro protocol for differentiating male rhesus (r) macaque PGCLCs (rPGCLCs) from induced pluripotent stem cells (riPSCs). Specifically, transplantation of aggregates containing rPGCLCs into mouse and nonhuman primate testicles overcomes a major bottleneck in rPGCLC differentiation. These findings suggest that immature rPGCLCs once transplanted into an adult gonadal niche commit to differentiate towards late rPGCs that initiate epigenetic reprogramming but do not complete the conversion into ENO2-positive spermatogonia.

Correction to: risk factors for cutaneous leishmaniasis in the rainforest of Bolivia: a cross-sectional study.

[This corrects the article DOI: 10.1186/s41182-018-0089-6.].

Risk factors for cutaneous leishmaniasis in the rainforest of Bolivia: a cross-sectional study.

BACKGROUND: Cutaneous leishmaniasis (CL) is an endemic disease in Bolivia, particularly in the rainforest of Cochabamba, in the municipality of Villa Tunari. The precarious, dispersed, and poorly accessible settlements in these farming communities make it difficult to study them, and there are no epidemiological studies in the area. The aim of the present study was to identify the risk factors associated with cutaneous leishmaniasis. METHODS: A cross-sectional study was conducted in August 2015 and August 2016 in two communities of Villa Tunari, Cochabamba. The cases were diagnosed through clinical examinations, identification of the parasite by microscopic examination, and the Montenegro skin test. Risk factors were identified through logistic regression. RESULTS: A total of 274 participants (40.9% female and 59.1% male) were surveyed, of which 43% were CL positive. Sex was the only factor associated with CL with three times more risk for men than for women; this finding suggests a sylvatic mechanism of transmission in the area. CONCLUSIONS: It is advisable to focus on education and prevention policies at an early age for activities related to either leisure or work. Further research is needed to assess the influence of gender-associated behavior for the risk of cutaneous leishmaniasis.

Assessment of a Leishmaniasis Reporting System in Tropical Bolivia Using the Capture-Recapture Method.

This study evaluates the level of underreporting of the National Program of Leishmaniasis Control (NPLC) in two communities of Cochabamba, Bolivia during the period 2013-2014. Montenegro skin test-confirmed cases of cutaneous leishmaniasis (CL) were identified through active surveillance during medical campaigns. These cases were compared with those registered in the NPLC by passive surveillance. After matching and cleaning data from the two sources, the total number of cases and the level of underreporting of the National Program were calculated using the capture-recapture analysis. This estimated that 86 cases of CL (95% confidence interval [CI]: 62.1-110.8) occurred in the study period in both communities. The level of underreporting of the NPLC in these communities was very high: 73.4% (95% CI: 63.1-81.5%). These results can be explained by the inaccessibility of health services and centralization of the NPLC activities. This information is important to establish priorities among policy-makers and funding organizations as well as implementing adequate intervention plans.

An integration-free, virus-free rhesus macaque induced pluripotent stem cell line (riPSC90) from embryonic fibroblasts.

The rhesus macaque induced pluripotent stem cell (riPSC) line, UCLAi090-A (riPSC90), was generated from rhesus embryonic fibroblast (REF) cells called REF90. REF90 cells and the riPSC90 line were authenticated by short tandem repeat analysis and had a normal male (42, XY) karyotype. The riPSC90 line expressed markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA4, and generated teratomas after transplantation into immunocompromised mice. riPSC90 could be used in parallel with riPSC89, which was derived from REFs cultured from a different rhesus macaque embryo (Sosa et al. 2016).

An integration-free, virus-free rhesus macaque induced pluripotent stem cell line (riPSC89) from embryonic fibroblasts.

We generated a rhesus macaque induced pluripotent stem cell (riPSC) line, riPSC89, from rhesus embryonic fibroblasts (REFs). Fibroblasts were expanded from the skin of a rhesus macaque embryo at embryonic day 47. REFs and riPSCs had a normal male (42, XY) karyotype. The riPSC89 line was positive for markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA4. Pluripotency was demonstrated through the generation of teratomas using transplantation into immunocompromised mice. The riPSC89 line may be a useful non-human primate resource to uncover developmental origins of disease, or used as a basic model to understand lineage specification in the primate embryo.

The immune response in patients with cutaneous leishmaniasis and the influence of zinc supplementation.

Cutaneous leishmaniasis triggers a varied immune response depending on parasite and host factors, which in turn can be influenced by nutrients. The resistance to the infection is associated with the Th1 type of cytokine production. The Th1 type can be reduced as a consequence of zinc deficiency, which may increase the risk for chronicity of the infection. Using in vitro and ex vivo models, we studied the influence of zinc supplementation on the immune response in patients with cutaneous leishmaniasis treated with antimony and the data were also compared to those of matched controls. Twenty-nine patients with cutaneous leishmaniasis (n=14 in zinc-supplemented group [45mg/day] and n=15 in placebo group) were treated by intramuscular injections of antimony for 20 days and took supplements for 60 days. Immunoglobulins in plasma and cell proliferation, IFN-γ production and CD markers of isolated peripheral blood mononuclear cells (PBMC) were measured. It was found that the cellular immune response of the patients maintained its activity as assessed by the ability of the PBMC to proliferate and produce IFN-γ in response to concanavalin A. Moreover, there was no difference in these variables between the zinc-supplemented and placebo groups after 60 days. The addition of zinc sulphate in vitro to PBMC reduced the IFN-γ production in the placebo group only. It is concluded that the cellular immune response of the cutaneous leishmaniasis patients remained active during treatment by antimony when compared to that of controls. It was not possible to document an additional effect of zinc supplementation for 60 days on the immune response.