Hematopoietic stem cell transplantation reverses white matter injury measured by diffusion-tensor imaging (DTI) in sickle cell disease patients.

Brain injury in sickle cell disease (SCD) comprises a wide spectrum of neurological damage. Neurocognitive deficits have been described even without established neurological lesions. DTI is a rapid, noninvasive, and non-contrast method that enables detection of normal-appearing white matter lesions not detected by conventional magnetic resonance imaging (MRI). The aim of the study was to evaluate if stem cell transplantation can revert white matter lesions in patients with SCD. Twenty-eight SCD patients were evaluated with MRI and DTI before and after allogeneic hematopoietic stem cell transplantation (HSCT), compared with 26 healthy controls (HC). DTI metrics included fractional anisotropy (FA), mean diffusivity (MD), radial (RD), and axial (AD) diffusivity maps, global efficiency, path length, and clustering coefficients. Compared to HC, SCD patients had a lower FA (p = 0.0086) before HSCT. After HSCT, FA increased and was not different from healthy controls (p = 0.1769). Mean MD, RD, and AD decreased after HSCT (p = 0.0049; p = 0.0029; p = 0.0408, respectively). We confirm previous data of white matter lesions in SCD and present evidence that HSCT promotes recovery of brain injury with potential improvement of brain structural connectivity.

Septic shock following hysteroscopy - A case report.

INTRODUCTION: Minimally invasive gynecological surgery such as hysteroscopy has a small risk of complications. These include uterine perforation (with or without adjacent pelvic organ lesion), bleeding and infection, and are more common in the presence of risk factors such as smoking, history of pelvic inflammatory disease (PID) and endometriosis. CASE PRESENTATION: A patient submitted to a diagnostic hysteroscopy with no immediate complications was admitted five days later to the emergency department in septic shock. The diagnosis of ruptured tubal abscess was made, requiring emergency laparotomy with sub-total hysterectomy and bilateral adnexectomy. Despite multiple organ failure requiring admission to the intensive care unit, the patient made a full recovery. CONCLUSION: Ascending infection can be a life-threatening complication of hysteroscopy, even in the absence of previously known risk factors.

Consensus Recommendations for the Use of Hyperdiluted Calcium Hydroxyapatite (Radiesse) as a Face and Body Biostimulatory Agent.

BACKGROUND: Calcium hydroxyapatite (CaHA) is a well-established collagen stimulator. In recent years, it has been increasingly used in hyperdiluted form as a biostimulatory agent rather than a volumizing filler to improve skin quality and firmness in both facial and corporal areas. However, guidelines on the techniques required to achieve optimal results are still lacking. The objective of this study was to develop a consensus recommendation for the safe and effective use of hyperdiluted CaHA for face and body biostimulation. METHODS: A team of 10 experts with extensive experience in dermal fillers and biostimulatory treatments for facial and body rejuvenation convened for a live meeting. Consensus was defined as approval by 70%-89% of all participants, whereas agreement of ≥90% denoted strong consensus. RESULTS: For most items, the group achieved a majority consensus. Recommendations have been provided for the face, neck, décolletage, buttocks, thighs, arms, abdomen, knees, and elbows with detailed injection techniques, providing information on insertion points, dosages, and volumes for both needle and cannula injections as well as the number of treatment sessions and intervals. CONCLUSIONS: The expert consensus supports and provides guidance for the use of CaHA as a biostimulatory agent for face and body rejuvenation. However, further clinical studies are necessary to provide physicians with the best evidence for the best treatment practices.

The crystal structure of the Leishmania infantum Silent Information Regulator 2 related protein 1: Implications to protein function and drug design.

The de novo crystal structure of the Leishmania infantum Silent Information Regulator 2 related protein 1 (LiSir2rp1) has been solved at 1.99Å in complex with an acetyl-lysine peptide substrate. The structure is broadly commensurate with Hst2/SIRT2 proteins of yeast and human origin, reproducing many of the structural features common to these sirtuin deacetylases, including the characteristic small zinc-binding domain, and the larger Rossmann-fold domain involved in NAD+-binding interactions. The two domains are linked via a cofactor binding loop ordered in open conformation. The peptide substrate binds to the LiSir2rp1 protein via a cleft formed between the small and large domains, with the acetyl-lysine side chain inserting further into the resultant hydrophobic tunnel. Crystals were obtained only with recombinant LiSir2rp1 possessing an extensive internal deletion of a proteolytically-sensitive region unique to the sirtuins of kinetoplastid origin. Deletion of 51 internal amino acids (P253-E303) from LiSir2rp1 did not appear to alter peptide substrate interactions in deacetylation assays, but was indispensable to obtain crystals. Removal of this potentially flexible region, that otherwise extends from the classical structural elements of the Rossmann-fold, specifically the ß8-ß9 connector, appears to result in lower accumulation of the protein when expressed from episomal vectors in L. infantum SIR2rp1 single knockout promastigotes. The biological function of the large serine-rich insertion in kinetoplastid/trypanosomatid sirtuins, highlighted as a disordered region with strong potential for post-translational modification, remains unknown but may confer additional cellular functions that are distinct from their human counterparts. These unique molecular features, along with the resolution of the first kinetoplastid sirtuin deacetylase structure, present novel opportunities for drug design against a protein target previously established as essential to parasite survival and proliferation.

Mini-Laparoscopy for Removal (Partial) of Adnexae at the Time of Hysterectomy.

STUDY OBJECTIVE: To demonstrate our technique of mini-laparoscopic adnexectomy or salpingectomy at the time of total laparoscopic hysterectomy (TLH). DESIGN: Step-by-step video demonstration of our technique. SETTING: The advantages of laparoscopic surgery have been widely recognized, including improved visualization and exposure, reduced operative trauma owing to smaller incisions and gentler tissue handling, and faster postoperative recovery. Continuing technological developments have allowed the use of smaller-caliber instruments while maintaining a high standard of surgical performance. Mini-laparoscopy requires the use of 3-mm or smaller ports. The main advantage of mini-laparoscopy is the reduced incision size, which can translate into a lower incidence of incision-related complications such as postoperative pain, infection, and trocar site herniation, along with superior cosmetic results. Today, in younger patients, prophylactic salpingectomy can be considered instead of adnexectomy, taking into account the well-known benefits of ovarian conservation. Prophylactic salpingectomy involves Fallopian tube removal for primary prevention of epithelial carcinoma of the fallopian tubes, ovaries, and peritoneum in women undergoing pelvic surgery for another indication. Other advantages of this intervention are the avoidance of hydrosalpinx (which affects ∼30% women after hysterectomy), the 7.8% lifetime risk of revision surgery [1], tubal infection, and benign and malignant Fallopian tube tumors. Finally, salpingectomy has no known physiological side effects, is safe and feasible, does not worsen surgical outcomes, does not significantly increase the operative time, and is not related to increased rates of intraoperative and postoperative complications or readmission. INTERVENTIONS: The patient is a 44-year-old woman with a history of 2 previous cesarean sections with adenomyosis and endometriosis infiltration of the uterosacral ligaments. After discussion about the risks and benefits of ovarian conservation with prophylactic salpingectomy versus adnexectomy, the patient opted to preserve her ovaries. A TLH with partial removal of the uterosacral ligaments nodules and prophylactic bilateral salpingectomy was performed. To begin, the patient was placed in lithotomy position with Allen stirrups at an angle of approximately 100 degrees. Standard trocar placement was used. A 5- or 10-mm 0° scope was placed at the level of the umbilicus and three 3-mm skin incisions were made for accessory lower quadrant trocar placement: 2 lateral, approximately 3 cm medial to the anterior superior iliac spine, and 1 suprapubic, slightly higher than the line made by the lateral trocars, ensuring that the distance between this port and the camera trocar exceeded 8 cm. This triangulation of the accessory ports allowed for good ergonomics for the surgeon. The procedure continued with abdominopelvic cavity inspection and bilateral transperitoneal ureter identification and eventual adhesiolysis, and then the following steps: The instruments used were a 10-mm scope, a 3-mm bipolar forceps, 3-mm cold scissors, a 3-mm suction-irrigation device and 3-mm grasping forceps. CONCLUSION: Mini-laparoscopy is an alternative to classic laparoscopy associated with greater patient satisfaction. Prophylactic salpingectomy has proven to reduce the risk of ovary, peritoneal, and tubal epithelial carcinomas as well as benign tubal diseases, and does not significantly increase the operative time or the incidence of postoperative complications.

Leishmania infantum Asparagine Synthetase A Is Dispensable for Parasites Survival and Infectivity.

A growing interest in asparagine (Asn) metabolism has currently been observed in cancer and infection fields. Asparagine synthetase (AS) is responsible for the conversion of aspartate into Asn in an ATP-dependent manner, using ammonia or glutamine as a nitrogen source. There are two structurally distinct AS: the strictly ammonia dependent, type A, and the type B, which preferably uses glutamine. Absent in humans and present in trypanosomatids, AS-A was worthy of exploring as a potential drug target candidate. Appealingly, it was reported that AS-A was essential in Leishmania donovani, making it a promising drug target. In the work herein we demonstrate that Leishmania infantum AS-A, similarly to Trypanosoma spp. and L. donovani, is able to use both ammonia and glutamine as nitrogen donors. Moreover, we have successfully generated LiASA null mutants by targeted gene replacement in L. infantum, and these parasites do not display any significant growth or infectivity defect. Indeed, a severe impairment of in vitro growth was only observed when null mutants were cultured in asparagine limiting conditions. Altogether our results demonstrate that despite being important under asparagine limitation, LiAS-A is not essential for parasite survival, growth or infectivity in normal in vitro and in vivo conditions. Therefore we exclude AS-A as a suitable drug target against L. infantum parasites.

Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

Prevalence and risk analysis for depression after open-heart valve replacement surgery.

BACKGROUND AND PURPOSE: Cardiac surgery may account for long term complications as depression. Geriatric Depression Scale (GDS) is a reliable 30-item questionnaire for assessing depressive symptoms in several populations. The aim of the present study was to determine the prevalence and predictors of depression after open-heart valve replacement surgery in a portuguese population. METHODS: Prospective observational study enrolling 52 patients (55.8% men, mean age 67.7 ± 11.3 years) with no history of depression, submitted to elective open-heart valve replacement surgery. Patients completed the GDS questionnaire at 6-month follow-up. Cardiovascular risk factors, medical history, left ventricular function, prescription table, analytical and surgical variables and length of hospital stay were collected to verify its influence on postoperative depression. RESULTS: Twenty-seven patients (41.4% men, mean age 69.3 ± 10.3 years) had scores consistent with symptoms of depression (GDS score >10), representing 51.9% of the sample. Twenty-one (77.8%) were mildly depressed, 6 (22.2%) were severely depressed and only 4 (19.0%) and 3 (50.0%) were on antidepressants, respectively. Postoperative complications (29.6 vs 8.0%, p=0.045), lower postoperative hemoglobin concentration (8.9 ± 0.8 vs 9.8 ± 1.5 g/dL p=0.015) and longer time of hospitalization (12.2 ± 7.7 vs 8.2 ± 3.4 days, p=0.020) were found to be significant predictors for postoperative depression. No other statistically significant differences were found. CONCLUSION: Depression after open-heart valve replacement surgery is frequent but appears to be generally overlooked. Strategies for systematic screening and early guidance should be implemented to ensure better health and quality-of-life of patients undergoing major cardiac surgery.

Knockdown of asparagine synthetase A renders Trypanosoma brucei auxotrophic to asparagine.

Asparagine synthetase (AS) catalyzes the ATP-dependent conversion of aspartate into asparagine using ammonia or glutamine as nitrogen source. There are two distinct types of AS, asparagine synthetase A (AS-A), known as strictly ammonia-dependent, and asparagine synthetase B (AS-B), which can use either ammonia or glutamine. The absence of AS-A in humans, and its presence in trypanosomes, suggested AS-A as a potential drug target that deserved further investigation. We report the presence of functional AS-A in Trypanosoma cruzi (TcAS-A) and Trypanosoma brucei (TbAS-A): the purified enzymes convert L-aspartate into L-asparagine in the presence of ATP, ammonia and Mg(2+). TcAS-A and TbAS-A use preferentially ammonia as a nitrogen donor, but surprisingly, can also use glutamine, a characteristic so far never described for any AS-A. TbAS-A knockdown by RNAi didn't affect in vitro growth of bloodstream forms of the parasite. However, growth was significantly impaired when TbAS-A knockdown parasites were cultured in medium with reduced levels of asparagine. As expected, mice infections with induced and non-induced T. brucei RNAi clones were similar to those from wild-type parasites. However, when induced T. brucei RNAi clones were injected in mice undergoing asparaginase treatment, which depletes blood asparagine, the mice exhibited lower parasitemia and a prolonged survival in comparison to similarly-treated mice infected with control parasites. Our results show that TbAS-A can be important under in vivo conditions when asparagine is limiting, but is unlikely to be suitable as a drug target.

Importância da variação do PASI realizado por diversos observadores / Inter-rater concordance study of the PASI (Psoriasis Area and Severity Index)

FUNDAMENTO: O índice da área e gravidade da psoríase foi método desenvolvido em 1978, para quantificar a gravidade da psoríase. Desde então, têm recebido algumas críticas, por ser longo, complexo e apresentar divergências na sua interpretação. OBJETIVO: Demonstrar o grau de concordância do índice da área e gravidade da psoríase aplicado em pacientes de psoríase por três avaliadores isoladamente. MATERIAL E MÉTODO: Estudo observacional, transversal, realizado no ambulatório de psoríase, no ano de 2007, onde foram selecionados 20 pacientes. Os pacientes foram avaliados por três pós-graduandos de dermatologia, que determinaram o índice da área e gravidade da psoríase, isoladamente. A fidedignidade interobservador foi avaliada pelo coeficiente de correlação intraclasses (ICC) e os gráficos de Bland & Altman. RESULTADOS: O ICC de 0,729 (IC 95 por cento: 0,440 - 0,882) demonstrou que existe boa concordância entre os observadores. Pela análise dos gráficos de Bland e Altman, no máximo, duas diferenças estão fora dos limites de concordância, expressando boa concordância. Observou-se também que existe comportamento sistemático nas diferenças do índice da área e gravidade da psoríase em relação a sua média, segundo o qual mostrou para valores altos do índice da área e gravidade da psoríase, ou seja, um predomínio de diferenças maiores. CONCLUSÃO: Notamos, através deste estudo, que o índice da área e gravidade da psoríase apresenta bons critérios de confiabilidade quando houve a avaliação da gravidade da psoríase, mesmo que realizado por diferentes avaliadores.

BACKGROUND: Developed in 1978, the PASI (Psoriasis Area and Severity Index) is a method to rate psoriasis severity. Since then, it has been criticized for being extremely long, complex and for yielding significantly divergent interpretations. OBJECTIVE: To determine the PASI inter-rater reliability among three independent evaluators examining the same patients. METHODS:Cross-sectional study conducted in 2007 at a psoriasis treatment center, where 20 patients were selected. The patients were evaluated by three postgraduate students of Dermatology who independently determined the PASI of each patient. The inter-rater reliability coefficient was determined by employing intraclass correlation coefficients (ICC) and Bland & Altman plots. RESULTS: An ICC of 0.729 (IC 95 percent: 0.440 - 0.882) showed a significant degree of concordance among evaluators. By graphic means, the Bland & Altman plot method presents concordance gap ranges. As shown in this study, narrow gap ranges indicate a good concordance degree. It was also observed that for high PASI values, wider concordance gap ranges predominate. CONCLUSION: Based on this analysis, we may conclude that the PASI is a reliable indicator of psoriasis severity because it shows significant concordance when independent evaluations are performed.

Simple and reproducible HPLC-DAD-ESI-MS/MS analysis of alkaloids in Catharanthus roseus roots.

Catharanthus roseus is one of the most important medicinal plants worldwide. The leaves of this species are the only source of the indolomonoterpenic alkaloids vincristin (leurocristine) and vinblastin (vincaleucoblastine), whose anticancer activity represents powerful therapeutics to many diseases, such as Hodgkin lymphoma. Usually, the remaining plant parts go to waste. Here we describe a phytochemical study on this species roots. Alkaloids in aqueous extracts, the usual form of consumption of this matrix, were studied using HPLC-DAD-ESI-MS/MS, which allowed the identification of 19-S-vindolinine, vindolinine, ajmalicine and an ajmalicine isomer, tabersonine, catharanthine, serpentine and a serpentine isomer. Quantification of the identified compounds revealed that serpentine and its isomer were predominant (64.7%) over the other alkaloids, namely vindolinine and its isomer (23.9%), catharanthine (7.7%) and ajmalicine (3.8%). The used procedure revealed to be simple, sensitive and reproducible.