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Abstract: Pesticides are commonly used worldwide and almost every human is potentially exposed to these chemicals. Exposure to pesticides such as permethrin and malathion has been associated with hematological malignancies in epidemiological studies. However, biological evidence showing if these chemicals induce genetic aberrations involved in the etiology of leukemia and lymphoma is missing. In our previous work, we have shown that a single high exposure (200 μ m, 24 hours) of permethrin and malathion induce damage in genes associated with hematological malignancies in peripheral blood mononuclear cells analyzed by interphase fluorescence in situ hybridization (FISH). In the present study, we assessed by FISH whether exposure to low concentrations (0.1 μ m, 72 hours) of permethrin and malathion induce aberrations in KMT2A and IGH genes, which are involved in the etiology of leukemia and lymphoma. Peripheral blood mononuclear cells were exposed to the chemicals, and damage in these genes was assessed on interphases and metaphases. We observed that both chemicals at low concentration induced structural aberrations in KMT2A and IGH genes. A higher level of damage was observed in KMT2A gene with malathion treatment and in IGH gene with permethrin exposure. We also observed numerical aberrations induced by these chemicals. The most frequent aberrations detected on interphase FISH were also observed on metaphases. Our results show that permethrin and malathion induce genetic damage in genes associated with hematological cancer, at concentrations biologically relevant. In addition, damage was observed on dividing cells, which suggests that these cells maintain their proliferation capacity in spite of the genetic damage they possess. [ABSTRACT FROM AUTHOR]

Rearrangements and overexpression of CRLF2 are hallmarks of poor outcomes in BCR::ABL1 -like B-ALL, and CRLF2 overexpression is a high-risk marker in T-ALL. However, CRLF2 alterations in pediatric hematologic malignancies other than B-ALL have not been reported. In this study, we analyzed the CRLF2 overexpression, rearrangements ( P2RY8::CRLF2 and IGH::CRLF2 ), activation (pSTAT5 and pERK), and the expression of dominant-negative IKZF1 isoforms (Ik6 and Ik8), implied in CRLF2 dysregulation, in 16 pediatric patients (AML, n  = 9; T-ALL, n  = 3; LBL, n  = 2; HL, n  = 1; cytopenia, n  = 1). A high frequency of CRLF2 rearrangements and overexpression was found in the 16 patients: 28.6% (4/14) showed CRLF2 overexpression, 93.8% (15/16) were positive for CRLF2 total protein (cell-surface and/or cytoplasmic), while 62.5% (10/16) were positive for P2RY8::CRLF2 and 12.6% (2/16) for IGH::CRLF2 . In addition, 43.8% (7/16) expressed Ik6 and Ik8 isoforms. However, only a few patients were positive for the surrogate markers pSTAT5 (14.3%; 2/14) and pERK (21.4%; 3/14).

Background: Computed tomography (CT) offers a detailed assessment of the shoulder for preoperative shoulder arthroplasty planning; however, this technique exposes the patient to ionizing radiation. The purpose of this study was to prospectively evaluate the practicality of reducing the CT radiation dose compared to conventional dose levels for manual and preoperative planning software measurements for shoulder arthroplasty., Methods: A total of 10 shoulder CT examinations were performed for preoperative planning purposes on a dual x-ray source CT scanner. A specialized dose-split scan technique was utilized to reconstruct CT images corresponding to 100%, 70%, and 30% radiation dose relative to our institution's standard of care imaging protocol. Glenoid version, inclination, and humeral head subluxation were measured manually by 3 authors and by commercially available software platforms. These measurements were analyzed for agreement among the 100%, 70%, and 30% dose levels for each patient. Tolerances of 5° of glenoid version, 5° of glenoid inclination, and 10% humeral head subluxation were used as equivalent for preoperative planning., Results: Automated measurements of 70% dose images were within 5° of version, 5° of inclination, and 10% subluxation in 95.0% of cases. Manual measurements of 70% RD images were within 5° of version for 90.0% of cases, 5° of inclination in 86.7% of cases, and 10% subluxation in 100% of cases. Automated measurements from the 30% dose images were within 5° of version, 5° of inclination, and 10% subluxation for 100% of cases. Manual measurements from the 30% dose images were within 5° of version for 86.7% of cases, 5° of inclination in 76.7% of cases, and 10% subluxation in 100% of cases. The mean absolute difference in software measurement of glenoid version (P = .96), glenoid inclination (P = .64), or humeral head subluxation (P = .09) or in aggregated manual mean absolute difference of version (P = .22), inclination (P = .31), or humeral head subluxation (P = .56) was not significant. Good to excellent reliability was determined by interclass correlation coefficients among the manual observers and automatic software platforms for measurements at all doses (P < .001) CONCLUSIONS: The results indicate that both preoperative planning software platforms and human observers produced similar measurements of glenoid version, inclination, and humeral head subluxation from reduced-dose images compared to standard of care doses. By implementing reduced dose techniques in preoperative shoulder CT, the potential risks associated with radiation exposure could be reduced for patients undergoing shoulder arthroplasty., (Copyright © 2024 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Purpose: The radial nerve may be painfully irritated or damaged by open reduction and internal fixation (ORIF) of humeral fractures. Secondary radial nerve lesions after ORIF of humeral shaft fractures are described in up to 16%. Not only peripheral nerves but also orthopaedic instruments and osteosynthesis material are well visible by ultrasound. The aim of this study was to evaluate the accuracy of ultrasound in assessing the relation between the bone overlapping screw tips and the radial nerve close to the humeral bone., Methods: Ultrasound-guided drilling was used to place screws as close as possible to the radial nerve in 8 humeral bones of four cadavers. The relation between the radial nerve and the screw tips was assessed by high-resolution ultrasound, and the overlap of all screw tips over the bone was measured by ultrasound and fluoroscopy. Thereafter, the findings were validated by anatomical dissection., Results: We could correctly identify all screw tips and their relation to the radial nerve by ultrasound. In 7 of 8 cases, the screw tip had direct contact with the radial nerve. The overlaying length of the screw tip was accurately measured by using ultrasound in all cases. In contrast fluoroscopy underestimated this length in 50% of cases., Conclusion: With this study, we show that ultrasound can reliable visualize the screw tips and its relation to the radial nerve. Ultrasound is a promising diagnostic tool to evaluate patients with radial nerve irritations or lesions after ORIF of humeral fractures. Furthermore, ultrasound could be an adequate tool to guide drilling., (© 2024. The Author(s).)

Background: Vasoocclusion in sickle cell disease can be precipitated by cold temperatures, hypoxia, infection, dehydration, and stress, all of which can occur in the setting of surgery. The purpose of this study was to identify predictors of perioperative complications among pediatric patients with sickle cell disease undergoing orthopaedic surgery., Methods: An institutional review board approved single-center retrospective review was conducted of pediatric patients 21 years of age and younger with SCD who underwent any orthopaedic surgery at a single center between 2009 and 2019. Patient data and procedure-specific information were recorded. Preoperative admission for hydration and/or blood transfusion and preoperative laboratory studies were reviewed. The primary study outcome was postoperative complications within 30 days of surgery requiring an ED visit or hospital admission., Results: Ninety-two patients who underwent 118 orthopaedic surgeries were identified. The average age at surgery was 12.0 years (SD 4.8 y). Surgical cases were classified as elective (n=82, 70%), infection (n=26, 22%), and trauma (n=9, 8%). The lower extremity was the most frequent surgical site (n=86, 73%). Sixty surgeries (51%) received a preoperative blood transfusion. There were 19 surgeries with postoperative complications (16%) that required an ED visit or hospital readmission within 30 days of surgery. There were significantly more complications following surgery on the hip as compared with other sites (24% vs. 9%, P =0.04). Four or more ED visits in the past year were associated with an OR of 5.7 for a postoperative complication ( P =0.01, 95% CI 1.6-20.5). Patients who had a preoperative blood transfusion had significantly greater rates of complications than those that did not (27% vs. 5%, P <0.01)., Conclusions: Children with SCD are at increased risk for complications after orthopaedic surgery, and the current study found an overall postoperative complication rate of 16%. Patients undergoing hip surgery had a disproportionate number of complications, with a 5.8-fold increased risk of a postoperative complication. Patients with 4 or more ED visits in the past year had a 5.7-fold increased risk of a complication., Level of Evidence: IV Retrospective case series., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

The evidence supporting the biological plausibility of the association of permethrin and malathion with hematological cancer is limited and contradictory; thus, further studies are needed. This study aimed to investigate whether in vitro exposure to 0.1 μM permethrin and malathion at 0, 24, 48 and 72 h after cell culture initiation induced changes in the gene expression and DNA methylation in mononuclear cells from bone marrow and peripheral blood (BMMCs, PBMCs). Both pesticides induced several gene expression modifications in both tissues. Through gene ontology analysis, we found that permethrin deregulates ion channels in PBMCs and BMMCs and that malathion alters genes coding proteins with nucleic acid binding capacity, which was also observed in PBMCs exposed to permethrin. Additionally, we found that both insecticides deregulate genes coding proteins with chemotaxis functions, ion channels, and cytokines. Several genes deregulated in this study are potentially associated with cancer onset and development, and some of them have been reported to be deregulated in hematological cancer. We found that permethrin does not induce DNA hypermethylation but can induce hypomethylation, and that malathion generated both types of events. Our results suggest that these pesticides have the potential to modify gene expression through changes in promoter DNA methylation and potentially through other mechanisms that should be investigated.

Background: Endothelial colony-forming cells (ECFCs) contribute to postnatal vasculogenesis. In venous thromboembolic disease (VTD), they are functionally abnormal and produce high concentrations of TNF-α., Objective: To analyze the TNF-α signaling pathway and its relationship with the expression of cell-cycle regulators., Methods: Mononuclear cells (MNCs) were collected from the peripheral blood of 20 healthy human volunteers (controls) and 30 patients with VTD matched by age (20-50 years) and sex to obtain ECFCs. We analyzed the relative quantification of the gene transcripts of TNF, NFkB1, PLAU, HMOX1, GSS, eNOS, CDKN1A, and CDKN1B through quantitative RT-PCR (qRT-PCR assays). Identification of NF-κB and activated targets of each pathway: NF-κB (Ser536); IκBα (Ser32/Ser36); p38 (Thr180/Tyr182) JNK (Thr183/Tyr185), p53 and cell-cycle regulators: p16, p18, p21, p27, p57, Cyclin D, Cyclin E, Cyclin A, Cyclin B, CDK2, CDK4; cell-cycle status was determined by KI-67 and 7-AAD. Cells were analyzed with flow cytometry and the FlowJo vX software., Results: In ECFCs from VTD patients, TNF-α receptor and NFkB were overexpressed and hyper-phosphorylated; eNOS and HMOX1 were down-regulated; cell-cycle regulators (p53, p18, p21) were elevated. In addition, the cell cycle was locked in the G2 phase., Conclusions: Our results strongly suggest that these molecular alterations in the pathway of TNF-α and cell cycle regulation induce endothelial dysfunction, reduced proliferation potential and vascular regeneration, and consequently, the occurrence of new thrombotic events., Competing Interests: Conflict of Interest The authors have no conflicts of interest., (Copyright © 2022. Published by Elsevier Inc.)

Chronic Myeloid Leukemia (CML) originates in a leukemic stem cell that resides in the bone marrow microenvironment, where they coexist with cellular and non-cellular elements. The vascular microenvironment has been identified as an important element in CML development since an increase in the vascularization has been suggested to be related with poor prognosis; also, using murine models, it has been reported that bone marrow endothelium can regulate the quiescence and proliferation of leukemic stem and progenitor cells. This observation, however, has not been evaluated in primary human cells. In this report, we used a co-culture of primitive (progenitor and stem) CML cells with endothelial colony forming cells (ECFC) as an in vitro model to evaluate the effects of the vascular microenvironment in the leukemic hematopoiesis. Our results show that this interaction allows the in vitro maintenance of primitive CML cells through an inflammatory microenvironment able to regulate the proliferation of progenitor cells and the permanence in a quiescent state of leukemic stem cells., Competing Interests: The authors confirm that there are no conflicts of interest.

Current antineoplastic agents present multiple disadvantages, driving an ongoing search for new and better compounds. Four lupane-type triterpenes, 3α,24-dihydroxylup-20(29)-en-28-oic acid ( 1 ), 3α,23-dihydroxy-30-oxo-lup-20(29)-en-28-oic acid ( 2 ), 3α,23- O -isopropylidenyl-3α,23-dihydroxylup-20(29)-en-28-oic acid ( 3 ), and 3α,23-dihydroxylup-20(29)-en-28-oic acid ( 4 ), previously isolated from Phoradendron wattii , were evaluated on two cell lines of chronic (K562) and acute (HL60) myeloid leukemia. Compounds 1 , 2 , and 4 decreased cell viability and inhibit proliferation, mainly in K562, and exhibited an apoptotic effect from 24 h of treatment. Of particular interest is compound 2 , which caused arrest in active phases (G2/M) of the cell cycle, as shown by in silico study of the CDK1/Cyclin B/Csk2 complex by molecular docking. This compound [3α,23-dihydroxy-30-oxo-lup-20(29)-en-28-oic acid] s a promising candidate for incorporation into cancer treatments and deserves further study.

Ph-like subtypes with CRLF2 abnormalities are frequent among Hispano-Latino children with pre-B ALL. Therefore, there is solid ground to suggest that this subtype is frequent in Mexican patients. The genomic complexity of Ph-like subtype constitutes a challenge for diagnosis, as it requires diverse genomic methodologies that are not widely available in diagnostic centers in Mexico. Here, we propose a diagnostic strategy for Ph-like ALL in accordance with our local capacity. Pre-B ALL patients without recurrent gene fusions (104) were classified using a gene-expression profile based on Ph-like signature genes analyzed by qRT-PCR. The expressions of the CRLF2 transcript and protein were determined by qRT-PCR and flow cytometry. The P2RY8::CRLF2 , IGH::CRLF2, ABL1/2 rearrangements, and Ik6 isoform were screened using RT-PCR and FISH. Surrogate markers of Jak2-Stat5/Abl/Ras pathways were analyzed by phosphoflow. Mutations in relevant kinases/transcription factors genes in Ph-like were assessed by target-specific NGS. A total of 40 patients (38.5%) were classified as Ph-like; of these, 36 had abnormalities associated with Jak2-Stat5 and 4 had Abl. The rearrangements IGH::CRLF2, P2RY8::CRLF2 , and iAMP21 were particularly frequent. We propose a strategy for the detection of Ph-like patients, by analyzing the overexpression/genetic lesions of CRLF2 , the Abl phosphorylation of surrogate markers confirmed by gene rearrangements, and Sanger sequencing.

Noninvasive Treatments for Acute and Chronic Back Pain Abstract. The therapy of back pain - especially the medication with opioids - can be challenging for the treating physician. Specific back pain can often be diagnosed by imaging and successfully treated by surgery or medication. In contrast, nonspecific back pain can be worsened by inappropriate imaging, questionable surgical indications and uncontrolled drug use. For the therapy of nonspecific back pain, maintaining daily activity and exercise therapy is central. Opioids are effective drugs for short-term use. However, long-term use often leads to opioid-induced hyperalgesia and hormonal dysfunction with decreased quality of life and libido. Furthermore, opioids can lead to abuse and addiction. After an ineffective treatment with non-opioids, opioids may be given for a limited time period (if possible shorter than four weeks) according to international guidelines.

Peripheral Regional Anaesthesia for Perioperative Analgesia Abstract. Peripheral regional anaesthesia is the actual gold standard of opioid-sparing perioperative analgesia and is mainly used for surgery of the shoulder, arm and leg. Well-trained anaesthesiologists are the prerequisite for the correct individual risk-benefit assessment and the performance of the nerve blocks using a combination of ultrasound guidance and peripheral nerve stimulation (dual guidance). The postoperative care of the patients requires trained staff.

The gene fusions BCR-ABL1, TCF3-PBX1, and ETV6-RUNX1 are recurrent in B-cell acute lymphoblastic leukemia (B-ALL) and are found with low frequency in coexistence with CRLF2 (cytokine receptor-like factor 2) rearrangements and overexpression. There is limited information regarding the CRLF2 abnormalities and dominant-negative IKZF1 isoforms associated with surrogate markers of Jak2, ABL, and Ras signaling pathways. To assess this, we evaluated 24 Mexican children with B-ALL positive for recurrent gene fusions at diagnosis. We found CRLF2 rearrangements and/or overexpression, dominant-negative IKZF1 isoforms, and surrogate phosphorylated markers of signaling pathways coexisting with recurrent gene fusions. All the BCR-ABL1 patients expressed CRLF2 and were positive for pCrkl (ABL); most of them were also positive for pStat5 (Jak2/Stat5) and negative for pErk (Ras). TCF3-PBX1 patients with CRLF2 abnormalities were positive for pStat5, most of them were also positive for pCrkl, and two patients were also positive for pErk. One patient with ETV6-RUNX1 and intracellular CRLF2 protein expressed pCrkl. In some cases, the activated signaling pathways were reverted in vitro by specific inhibitors. We further analyzed a TCF3-PBX1 patient at relapse, identifying a clone with the recurrent gene fusion, P2RY8-CRLF2, rearrangement, and phosphorylation of the three surrogate markers that we studied. These results agree with the previous reports regarding resistance to treatment observed in patients with recurrent gene fusions and coexisting CRLF2 gene abnormalities. A marker phosphorylation signature was identified in BCR-ABL1 and TCF3-PBX1 patients. To obtain useful information for the assessment of treatment in B-ALL patients with recurrent gene fusions, we suggest that they should be evaluated at diagnosis for CRLF2 gene abnormalities and dominant-negative IKZF1 isoforms, in addition to the analyses of activation and inhibition of signaling pathways., (© 2021 The Authors. The Journal of Pathology: Clinical Research published by The Pathological Society of Great Britain and Ireland & John Wiley & Sons, Ltd.)

Asclepias linaria Cav. (Apocynaceae) is a shrubby plant endemic of Mexico which has been used in traditional medicine. However, the bioactive potential of this plant remains unexplored. In this study, the phenolic composition, antioxidant, and cytotoxic activities of A. linaria leaves were determined. In order to estimate the phenolic composition of the leaves, the total phenolic, flavonoid, and condensed tannins contents were determined. Furthermore, the antioxidant activity was measured by the scavenging activity of the 2,2-diphenyl-1-picrylhydrazyl (DPPH • ) and 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulphonic acid] (ABTS •+ ) radicals and the total antioxidant capacity. The phenolic compounds identified in the A. linaria leaves by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS) include phenolic acids, such as p -coumaric and ferulic acid, as well as flavonoids, such as rutin and quercetin. The leaves' extracts of A. linaria showed a high scavenging activity of DPPH • and ABTS •+ radicals (IC 50 0.12 ± 0.001 and 0.51 ± 0.003 µg/mL, respectively), high total antioxidant capacity values (99.77 ± 4.32 mg of ascorbic acid equivalents/g of dry tissue), and had a cytotoxic effect against K562 and HL60 hematologic neoplasia cells lines, but no toxicity towards the normal mononuclear cell line was observed. These results highlight the potential of A. linaria and could be considered as a possible alternative source of anticancer compounds.

Background: This study evaluated the feasibility and reliability of high-resolution ultrasonography (HRUS) of the radial nerve in the early, postoperative period after operative stabilization of humeral shaft fractures., Methods: This study enrolled patients between September 2015 and April 2018 with a humeral shaft fracture who were assessed with HRUS within 2 weeks after surgery. Based on the ultrasound artifacts, the examiners subjectively defined quality of ultrasound as "bad" or "good." The cross-sectional area of the radial and the posterior interosseous nerve was recorded at predefined locations. The radial nerve was scanned axially in the whole course to identify nerve continuity., Results: Of 44 patients who underwent operations for humeral shaft fracture, HRUS was used to assess 15 patients at an average 4.8 ± 2.6 days (range, 2-11 days) after surgery. The examiners defined ultrasound quality as "good" in 13 of 15 patients (~87%). Primary radial nerve palsy (RNP) was identified in 3 of the 15 patients, and 4 sustained secondary RNP. Nerve continuity was demonstrated by HRUS in every patient. In patients with RNP, nerve continuity was secondarily confirmed by surgical exploration or functional and electrophysiological recovery., Conclusion: Early postoperative HRUS of the radial nerve after osteosynthesis of humeral shaft fractures is a feasible and reliable method to identify radial nerve continuity. In case of pathology, this assessment tool can additionally provide valuable information concerning location and etiology of the RNP., (Copyright © 2018 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Tyrosine kinase inhibitors (TKI) have become a first-line treatment for chronic myeloid leuakemia (CML). TKIs efficiently target bulk CML cells; however, they are unable to eliminate the leukaemic stem cell (LSC) population that causes resistance and relapse in CML patients. In this study, we assessed the effects of parthenolide (PTL) and dimethyl amino parthenolide (DMAPT), two potent inhibitors of LSCs in acute myeloid leukaemia (AML), on CML bulk and CML primitive (CD34 + lin - ) cells. We found that both agents induced cell death in CML, while having little effect on the equivalent normal hematopoietic cells. PTL and DMAPT caused an increase in reactive oxygen species (ROS) levels and inhibited NF-κB activation. PTL and DMAPT inhibited cell proliferation and induced cell cycle arrest in G 0 and G 2 phases. Furthermore, we found cell cycle inhibition to correlate with down-regulation of cyclin D1 and cyclin A. In summary, our study shows that PTL and DMAPT have a strong inhibitory effect on CML cells. Given that cell cycle arrest was not dependent on ROS induction, we speculate that this effect could be a direct consequence of NF-κB inhibition and if this mechanism was to be evaded, PTL and DMAPT induced cell death would be potentiated., (© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

In this study, we determined the gene expression profiles of bone marrow-derived cell fractions, obtained from normal subjects and Chronic Myeloid Leukemia (CML) patients, that were highly enriched for hematopoietic stem (HSCs) and progenitor (HPCs) cells. Our results indicate that the profiles of CML HSCs and HPCs were closer to that of normal progenitors, whereas normal HSCs showed the most different expression profile of all. We found that the expression profiles of HSCs and HPCs from CML marrow were closer to each other than those of HSCs and HPCs from normal marrow. The major biologic processes dysregulated in CML cells included DNA repair, cell cycle, chromosome condensation, cell adhesion, and the immune response. We also determined the genomic changes in both normal and CML progenitor cells under culture conditions, and found that several genes involved in cell cycle, steroid biosynthesis, and chromosome segregation were upregulated, whereas genes involved in transcription regulation and apoptosis were downregulated. Interestingly, these changes were the same, regardless of the addition of Imatinib (IM) to the culture. Finally, we identified three genes-PIEZO2, RXFP1, and MAMDC2- that are preferentially expressed by CML primitive cells and that encode for cell membrane proteins; thus, they could be used as biomarkers for CML stem cells., (© 2017 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Background: Venous thromboembolic disease (VTD) is a public health problem. We recently reported that endothelial colony-forming cells (ECFCs) derived from endothelial cells (EC) (ECFC-ECs) from patients with VTD have a dysfunctional state. For this study, we proposed that a dysfunctional status of these cells generates a reduction of its proliferative ability, which is also associated with senescence and reactive oxygen species (ROS)., Methods and Results: Human mononuclear cells (MNCs) were obtained from peripheral blood from 40 healthy human volunteers (controls) and 50 patients with VTD matched by age (20-50 years) and sex to obtain ECFCs. We assayed their proliferative ability with plasma of patients and controls and supernatants of cultures from ECFC-ECs, senescence-associated β-galactosidase (SA-β-gal), ROS, and expression of ephrin-B2/Eph-B4 receptor. Compared with cells from controls, cells from VTD patients showed an 8-fold increase of ECFCs that emerged 1 week earlier, reduced proliferation at long term (39%) and, in passages 4 and 10, a highly senescent rate (30±1.05% vs. 91.3±15.07%, respectively) with an increase of ROS and impaired expression of ephrin-B2/Eph-4 genes. Proliferation potential of cells from VTD patients was reduced in endothelial medium [1.4±0.22 doubling population (DP)], control plasma (1.18±0.31 DP), or plasma from VTD patients (1.65±0.27 DP)., Conclusions: As compared with controls, ECFC-ECs from individuals with VTD have higher oxidative stress, proliferation stress, cellular senescence, and low proliferative potential. These findings suggest that patients with a history of VTD are ECFC-ECs dysfunctional that could be associated to permanent risk for new thrombotic events.

Objective: Intrathecal drug delivery (ITDD) is commonly used for intractable pain management. A paucity of good-quality studies in chronic noncancer patients and concerns over increased dosages have focused interest on different modes of administration. The aim of this international multicenter randomized double-blind crossover trial was to compare the efficacy of the same daily dose of drugs administered by intermittent boluses vs simple continuous infusion., Methods: Eligible patients implanted with a programmable ITDD device were randomized to receive two weeks of either intermittent boluses or a simple continuous flow in period 1, followed by a crossover to the alternative mode of administration. The primary outcome measure was the Patients' Global Impression of Change (PGIC) scale., Results: The mean proportion of positive responders (at least "minimally improved") was 38.4% in the continuous condition vs 37.3% in the bolus (difference in proportions = 1.1%, 95% confidence interval [CI] = -21.8-24.0%, P  = 0.93). The mean PGIC in the continuous condition was 3.8 vs 3.9 in the bolus (mean difference = -0.1, -0.6-0.4, P  = 0.72). Exploratory analyses revealed a tendency for the mean proportion of positive responders to be higher at low vs high flow rates for both bolus and continuous administrations. Two patients were withdrawn from the study due to adverse events during the bolus phase, both with symptoms of increased pain, and one patient with additional symptoms of numbness and urinary retention., Conclusion: The mean PGIC and proportion of positive responders was not substantially different after intermittent bolus vs continuous administration., (© 2016 American Academy of Pain Medicine. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com)

Several novel compounds have been developed for the treatment of different types of leukemia. In the present study, we have assessed the in vitro effects of Casiopeina III-Ea, a copper-containing small molecule, on cells from patients with Chronic Myeloid Leukemia (CML). We included primary CD34 + Lineage-negative (Lin - ) cells selected from CML bone marrow, as well as the K562 and MEG01 cell lines. Bone marrow cells obtained from normal individuals - both total mononuclear cells as well as CD34 + Lin - cells- were used as controls. IC50 corresponded to 0.5μM for K562 cells, 0.63μM for MEG01 cells, 0.38μM for CML CD34 + lin - cells, and 1.0μM for normal CD34 + lin - cells. Proliferation and expansion were also inhibited to significantly higher extents in cultures of CML cells as compared to their normal counterparts. All these effects seemed to occur via a bcr-abl transcription-independent mechanism that involved a delay in cell division, an increase in cell death, generation of Reactive Oxygen Species and changes in cell cycle. Our results demonstrate that Casiopeina III-Ea possesses strong antileukemic activity in vitro, and warrant further preclinical (animal) studies to assess such effects in vivo., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Chronic Myeloid Leukemia (CML) is sustained by a small population of cells with stem cell characteristics known as Leukemic Stem Cells that are positive to BCR-ABL fusion protein, involved with several abnormalities in cell proliferation, expansion, apoptosis and cell cycle regulation. Current treatment options for CML involve the use of Tirosine Kinase Inhibitor (Imatinib, Nilotinib and Dasatinib), that efficiently reduce proliferation proliferative cells but do not kill non proliferating CML primitive cells that remain and contributes to the persistence of the disease. In order to understand the role of Cyclin Dependent Kinase Inhibitors in CML LSC permanence after TKI treatment, in this study we analyzed cell cycle status, the levels of several CDKIs and the subcellular localization of such molecules in different CML cell lines, as well as primary CD34(+)CD38(-)lin(-) LSC and HSC. Our results demonstrate that cellular location of p18(INK4c) and p57(Kip2) seems to be implicated in the antiproliferative activity of Imatinib and Dasatinib in CML cells and also suggest that the permanence of quiescent stem cells after TKI treatment could be associated with a decrease in p18(INK4c) and p57(Kip2) nuclear location. The differences in p18(INK4c)and p57(Kip2)activities in CML and normal stem cells suggest a different cell cycle regulation and provide a platform that could be considered in the development of new therapeutic options to eliminate LSC.

Purpose: The goal of our study was to determine the precise femoral drill guide placement during reconstruction of the anterolateral bundle (ALB) of the posterior cruciate ligament (PCL) femoral footprint that would produce a minimum tunnel length of 25 mm, a maximum graft/femoral tunnel angle of 50°, and a minimum distance of 10 mm between the femoral socket and the subchondral bone of the weight-bearing surface of the medial femoral condyle., Methods: Using computer navigation, we used synthetic replicas of human femora to create a series of virtual femoral sockets. We then measured the bone tunnel length, angle of the femoral socket relative to the PCL footprint, and distance from the subchondral bone of the weight-bearing surface of the medial femoral condyle to the femoral socket at a series of guide pin sleeve positions. We positioned the guide pin using the following angle combinations: -20°, -10°, 0°, 10°, 20°, 30°, 40°, 50°, and 60° to a line perpendicular to the femoral axis in the coronal plane and -15°, 0°, 15°, 30°, 45°, and 60° to a line parallel to the transepicondylar axis in the axial plane. Using linear regression models, we determined the precise drill guide placement angles that would produce the optimal tunnel length, graft/femoral tunnel angle, and distance to the subchondral bone margin., Results: The results were consistent between small, medium, and large femora. We found that the optimal drilling angles for anatomic reconstruction of the femoral footprint of the ALB of the PCL were 0° to a line perpendicular to the femoral axis in the coronal plane and 15° to a line parallel to the transepicondylar axis in the horizontal or axial plane., Conclusions: During outside-in drilling for PCL reconstruction, holding the guide pin sleeve at a position 0° to a line perpendicular to the femoral axis in the coronal plane and 15° to a line parallel to the transepicondylar axis in the horizontal or axial plane results in optimal bone tunnel length, graft/tunnel angle, and distance between the femoral socket and the subchondral bone of the weight-bearing surface of the medial femoral condyle., Clinical Relevance: We describe a precise femoral tunnel drill guide placement during outside-in PCL reconstruction that ensures an optimal femoral socket with a minimum bone tunnel length of 25 mm, maximum graft/femoral tunnel angle of 50°, and minimum distance of 10 mm between the subchondral bone of the weight-bearing surface of the medial femoral condyle and the femoral socket., (Copyright © 2015 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Background and Aims: In trying to contribute to our knowledge on the biology of hematopoietic stem cells (HSC) and hematopoietic progenitor cells (HPC) from pediatric acute myeloid leukemia (AML), in the present study we analyzed the expression of four cell surface antigens relevant to human hematopoiesis-CD90, CD96, CD117, and CD123-in bone marrow from pediatric AML patients and normal control subjects., Methods: CD34(+) CD38(-) cells (enriched for HSC) and CD34(+) CD38(+) cells (enriched for HPC) were resolved on the basis of CD34 and CD38 expression. Concomitantly, expression of CD90 and CD96 or CD117 and CD123 was assessed by multicolor flow cytometry in each cell population., Results: CD90 and CD117 were expressed in a low proportion of CD34(+) CD38(-) and CD34(+) CD38(+) cells and no significant differences were observed between normal marrow and AML at diagnosis. In contrast, CD96(+) cells and CD123(+) cells were found at significantly higher levels in both cell populations from AML at diagnosis, as compared to normal marrow. Levels of both cell surface markers after treatment remained higher than in normal marrow., Discussion: These results show an increased frequency of CD96(+) and CD123(+) cells within the CD34(+) cell population from pediatric AML; this is consistent with the findings reported previously for adult AML. Our study supports the notion that expression of such antigens should be explored for their use as markers for diagnosis and prognosis., (Copyright © 2014 IMSS. Published by Elsevier Inc. All rights reserved.)

Summary: Non-ribosomal peptide synthetases (NRPSs) are multi-modular enzymes, which biosynthesize many important peptide compounds produced by bacteria and fungi. Some studies have revealed that an individual domain within the NRPSs shows significant substrate selectivity. The discovery and characterization of non-ribosomal peptides are of great interest for the biotechnological industries. We have applied computational mining methods in order to build a database of NRPSs modules that bind to specific substrates. We have used this database to build a hidden Markov model predictor of substrates that bind to a given NRPS., Availability: The database and the predictor are freely available on an easy-to-use website at www.nrpssp.com., Contact: carlos.prieto@unileon.es, Supplementary Information: Supplementary data is available at Bioinformatics online.

Widespread contamination of groundwater by methyl tertiary butyl ether (MTBE) has triggered the exploration of different technologies for in situ removal of the pollutant, including biostimulation of naturally occurring microbial communities or bioaugmentation with specific microbial strains known to biodegrade the oxygenate. After laboratory studies revealed that bacterial strain PM1 rapidly and completely biodegraded MTBE in groundwater sediments, the organism was tested in an in situ field study at Port Hueneme Naval Construction Battalion Center in Oxnard, California. Two pilot test plots (A and B) in groundwater located down-gradient from an MTBE source were intermittently sparged with pure oxygen. Plot B was also inoculated with strain PM1. MTBE concentrations up-gradient from plots A and B initially varied temporally from 1.5 to 6 mg MTBE/L. Six months after treatment began, MTBE concentrations in monitoring wells down-gradient from the injection bed decreased substantially in the shallow zone of the groundwater in plots A and B, thus even in the absence of the inoculated strain PM1. In the deeper zone, downstream MTBE concentrations also decreased in plot A and to a lesser extent in plot B. Difficulties in delivery of oxygen to the deeper zone of plot B, evidenced by low dissolved oxygen concentrations, were likely responsible for low rates of MTBE removal at that location. We measured the survival and movement of strain PM1 in groundwater samples using two methods for detection of DNA sequences specific to strain PM1: TaqMan quantitative polymerase chain reaction, and internal transcribed spacer region analysis. A naturally occurring bacterial strain with > 99% 16S rDNA sequence similarity to strain PM1 was detected in groundwater collected at various locations at Port Hueneme, including outside the plots where the organism was inoculated. Addition of oxygen to naturally occurring microbial populations was sufficient to stimulate MTBE removal at this site. In some cases, however, inoculation with an MTBE-degrading culture may be warranted.