Chiral two bladed ML2 metallamacrocycles: design, structures and solution behavior.
Chiral two bladed complexes of [Co(L)2][BF4]2 (1) and [Zn(L)2][BF4]2 (2) containing an atropoisomeric semi-rigid bidentate ligand L were synthesized and characterized. They are obtained as homochiral species but in a racemic mixture via a ligand self-sorting mechanism. These enantiomers can be differentiated in solution using optically active anions.


Biomedical system based on the Discrete Hidden Markov Model using the Rocchio-Genetic approach for the classification of internal carotid artery Doppler signals.
When the maximum likelihood approach (ML) is used during the calculation of the Discrete Hidden Markov Model (DHMM) parameters, DHMM parameters of the each class are only calculated using the training samples (positive training samples) of the same class. The training samples (negative training samples) not belonging to that class are not used in the calculation of DHMM model parameters. With the aim of supplying that deficiency, by involving the training samples of all classes in calculating processes, a Rocchio algorithm based approach is suggested. During the calculation period, in order to determine the most appropriate values of parameters for adjusting the relative effect of the positive and negative training samples, a Genetic algorithm is used as an optimization technique. The purposed method is used to classify the internal carotid artery Doppler signals recorded from 136 patients as well as of 55 healthy people. Our proposed method reached 97.38% classification accuracy with fivefold cross-validation (CV) technique. The classification results showed that the proposed method was effective for the classification of internal carotid artery Doppler signals.


Recurrence of granular corneal dystrophy type I deposits within host stroma after non-descemet baring anterior lamellar keratoplasty.
The aim of this study was to describe a case of recurrent granular deposits after non-Descemet baring anterior lamellar keratoplasty (nDALK). A 28-year-old male with granular corneal dystrophy type I, found to have deposits throughout the anterior and midstroma, underwent nDALK. Three years later, he had a recurrence of the deposits. Slit-lamp photographs and optical coherence tomography were used to document the level of recurrence. Full-thickness penetrating keratoplasty was performed, and the residual host stroma was sent for pathology. Slit-lamp photographs and anterior segment optical coherence tomography confirmed that the recurrent hyaline deposits were confined to the residual host stroma just anterior to Descemet membrane. The anterior lamellar graft and epithelium remained clear. Pathology showed positive staining with Masson trichrome in the host stroma just anterior to Descemet membrane. Previous studies suggest that the recurrence of granular deposits after keratoplasty is usually anterior and may even be epithelial in origin. This case report documents the recurrence of granular dystrophy entirely within the residual host stroma after nDALK. These findings suggest that residual keratocytes may still be a source of recurrence.


Diversity considerations in HIV-1 vaccine selection.
Globally, human immunodeficiency virus-type 1 (HIV-1) is extraordinarily variable, and this diversity poses a major obstacle to AIDS vaccine development. Currently, candidate vaccines are derived from isolates, with the hope that they will be sufficiently cross-reactive to protect against circulating viruses. This may be overly optimistic, however, given that HIV-1 envelope proteins can differ in more than 30% of their amino acids. To contend with the diversity, country-specific vaccines are being considered, but evolutionary relationships may be more useful than regional considerations. Consensus or ancestor sequences could be used in vaccine design to minimize the genetic differences between vaccine strains and contemporary isolates, effectively reducing the extent of diversity by half.


DYNAMIC AND STATIC VESSEL ANALYSIS IN PATIENTS WITH RETINITIS PIGMENTOSA: A Pilot Study of Vascular Diameters and Functionality.
To evaluate in vivo the vascular anatomy and functionality of early manifestation retinitis pigmentosa (RP) by means of a dynamic and static vessel analyzer. Fourteen patients with early RP and 14 normal subjects were consecutively enrolled in this observational, prospective study. Each patient underwent a complete ophthalmologic examination, including dynamic and static retinal vessel analysis using the Dynamic Vessel Analyzer. The patients with RP and the control group were well matched in age and sex. Patients with RP had a mean best-corrected visual acuity of 20/25 (range: 20/40-20/20). Dynamic vessel analysis performed in patients with RP showed an arterial and venous dilation during flicker stimulation of 5.28 ± 1.7% and 4.07 ± 1.78%, respectively. Only arterial dilation was statistically different compared with control subjects (3.33 ± 0.99%, P = 0.0062). Static retinal vessel analysis in patients with RP showed a decreased mean central retinal artery equivalent (P < 0.001) and central retinal vein equivalent (P < 0.001) compared with control subjects. By contrast, the arterial-to-venous ratio was similar in both groups (RP: 0.79 ± 0.11, control group 0.86 ± 0.04, P = 0.072). Our data confirm that retinal arterial and venous narrowing is present at an early stage in patients affected by RP. However, dynamic vessel analysis shows how the retina of patients with RP with no best-corrected visual acuity loss presents an augmented artery dilation response compared with normal subjects and retained neurovascular coupling.


Prognostic Communication Between Oncologists and Parents of Children With Advanced Cancer.
Parents of children with cancer perceive deficits in quality of prognostic communication. How oncologists disclose information about disease progression and incurability and how prognostic communication impacts parental understanding of prognosis are poorly understood. In this study, we aimed to (1) characterize communication strategies used by pediatric oncologists to share prognostic information across a child's advancing illness course and (2) explore relationships between different communication approaches and concordance of oncologist-parent prognostic understanding. In this prospective, longitudinal, mixed-methods study, serial disease reevaluation conversations were audio recorded across an advancing illness course for children with cancer and their families. Surveys and interviews also were conducted with oncologists and caregivers at specific time points targeting disease progression. Seventeen children experienced advancing illness on study, resulting in 141 recordings (40 hours). Fewer than 4% of recorded dialogue constituted prognostic communication, with most codes (77%) occurring during discussions about frank disease progression. Most recordings at study entry contained little or no prognosis communication dialogue, and oncologists rated curability lower than parents across all dyads. Parent-oncologist discordance typically was preceded by conversations without incurability statements; ultimately, concordance was achieved in most cases after the oncologist made direct statements about incurability. Content analysis revealed 3 distinct patterns (absent, deferred, and seed planting) describing the provision of prognostic communication across an advancing pediatric cancer course. When oncologists provided direct statements about incurability, prognostic understanding appeared to improve. Further research is needed to determine optimal timing for prognostic disclosure in alignment with patient and family preferences.


Inducible neuronal inactivation of Sim1 in adult mice causes hyperphagic obesity.
Germline haploinsufficiency of human or mouse Sim1 is associated with hyperphagic obesity. Sim1 encodes a transcription factor required for proper formation of the paraventricular (PVN), supraoptic, and anterior periventricular hypothalamic nuclei. Sim1 expression persists in these neurons in adult mice, raising the question of whether it plays a physiologic role in regulation of energy balance. We previously showed that Sim1 heterozygous mice had normal numbers of PVN neurons that were hyporesponsive to melanocortin 4 receptor agonism and showed reduced oxytocin expression. Furthermore, conditional postnatal neuronal inactivation of Sim1 also caused hyperphagic obesity and decreased hypothalamic oxytocin expression. PVN projections to the hindbrain, where oxytocin is thought to act to modulate satiety, were anatomically intact in both Sim1 heterozygous and conditional knockout mice. These experiments provided evidence that Sim1 functions in energy balance apart from its role in hypothalamic development but did not rule out effects of Sim1 deficiency on postnatal hypothalamic maturation. To address this possibility, we used a tamoxifen-inducible, neural-specific Cre transgene to conditionally inactivate Sim1 in adult mice with mature hypothalamic circuitry. Induced Sim1 inactivation caused increased food and water intake and decreased expression of PVN neuropeptides, especially oxytocin and vasopressin, with no change in energy expenditure. Sim1 expression was not required for survival of PVN neurons. The results corroborate previous evidence that Sim1 acts physiologically as well as developmentally to regulate body weight. Inducible knockout mice provide a system for studying Sim1's physiologic function in energy balance and identifying its relevant transcriptional targets in the hypothalamus.


Safe and Appropriate Use of Methadone in Hospice and Palliative Care: Expert Consensus White Paper.
Methadone has several unique characteristics that make it an attractive option for pain relief in serious illness, but the safety of methadone has been called into question after reports of a disproportionate increase in opioid-induced deaths in recent years. The American Pain Society, College on Problems of Drug Dependence, and the Heart Rhythm Society collaborated to issue guidelines on best practices to maximize methadone safety and efficacy, but guidelines for the end-of-life scenario have not yet been developed. A panel of 15 interprofessional hospice and palliative care experts from the U.S. and Canada convened in February 2015 to evaluate the American Pain Society methadone recommendations for applicability in the hospice and palliative care setting. The goal was to develop guidelines for safe and effective management of methadone therapy in hospice and palliative care. This article represents the consensus opinion of the hospice and palliative care experts for methadone use at end of life, including guidance on appropriate candidates for methadone, detail in dosing, titration, and monitoring of patients' response to methadone therapy.


Accurate localization of intracavitary brachytherapy applicators from 3D CT imaging studies.
To present an accurate method to identify the positions and orientations of intracavitary (ICT) brachytherapy applicators imaged in 3D CT scans, in support of Monte Carlo photon-transport simulations, enabling accurate dose modeling in the presence of applicator shielding and interapplicator attenuation. The method consists of finding the transformation that maximizes the coincidence between the known 3D shapes of each applicator component (colpostats and tandem) with the volume defined by contours of the corresponding surface on each CT slice. We use this technique to localize Fletcher-Suit CT-compatible applicators for three cervix cancer patients using post-implant CT examinations (3 mm slice thickness and separation). Dose distributions in 1-to-1 registration with the underlying CT anatomy are derived from 3D Monte Carlo photon-transport simulations incorporating each applicator's internal geometry (source encapsulation, high-density shields, and applicator body) oriented in relation to the dose matrix according to the measured localization transformations. The precision and accuracy of our localization method are assessed using CT scans, in which the positions and orientations of dense rods and spheres (in a precision-machined phantom) were measured at various orientations relative to the gantry. Using this method, we register 3D Monte Carlo dose calculations directly onto post insertion patient CT studies. Using CT studies of a precisely machined phantom, the absolute accuracy of the method was found to be +/-0.2 mm in plane, and +/-0.3 mm in the axial direction while its precision was +/-0.2 mm in plane, and +/-0.2 mm axially. We have developed a novel, and accurate technique to localize intracavitary brachytherapy applicators in 3D CT imaging studies, which supports 3D dose planning involving detailed 3D Monte Carlo dose calculations, modeling source positions, shielding and interapplicator shielding, accurately.


Human papillomavirus screening and cervical cancer prevention.
Infection with one of several types of human papillomavirus (HPV) appears to be a necessary first step in the development of invasive cervical cancer. We cannot currently treat HPV infections; thus, the role of HPV testing is to identify women with precancerous lesions that can be removed and, in so doing, prevent progression to invasive carcinoma. Although HPV testing may help to identify women at risk of cervical cancer who might be missed by other screening tests, it is inherently nonspecific at identifying those who would otherwise develop cervical cancer. In order to avoid overtreatment of women with minor lesions with little potential for progression, HPV testing needs to be repeated or combined with Pap smears. Protocols for HPV screening have yet to be properly evaluated. Here we consider several possible applications of HPV testing in the prevention of cervical cancer. The most immediate role is as a secondary test in women with minor cytological abnormalities. Appropriate use of HPV testing as a primary screening tool depends on the setting. In a developed country without an organized screening program, HPV testing might be used in addition to Pap smears in women age 35 and over to increase sensitivity. Within an organized screening program, HPV testing might be used in combination with Pap testing, but with extended screening intervals so as to obtain the maximum advantage to women without unduly increasing costs. Where resources are strictly limited, an attractive option would be to perform visual inspection of the cervix after application of dilute acetic acid using a low threshold for referral, and to test for HPV only on those with abnormal looking lesions.


Synergistic mediation of tumor signaling pathways in hepatocellular carcinoma therapy via dual-drug-loaded pH-responsive electrospun fibrous scaffolds.
With the understanding of tumorigenesis, the tumor-infiltrating inflammatory cells, which could contribute to carcinogenesis, have greatly drawn public attention. We hypothesized that controlling inflammation in the initial stage along with antitumor activity for a relatively long term could prevent tumor recurrence. Herein, we designed a novel electrospun composite poly(l-lactide) (PLLA) fibrous scaffold, which contained sodium bicarbonate (SB) and doxorubicin (DOX) inside mesoporous silica particles (MSNs) to achieve long-term pH-sensitive DOX release and ibuprofen (IBU) outside MSNs to achieve initial short-term release. This construct was found to exhibit an initial burst release of IBU at an early stage and sustained the release of DOX at a relatively later stage. In vivo results showed that when the fibrous scaffold was implanted in a liver-tumor-bearing nude mouse, the mouse lifespan was prolonged to 1.5 times higher than that when implanted with scaffolds with no IBU or SB. Moreover, the residual tumor treated with PLLA-(MSN/DOX-SB)-IBU fibers demonstrated considerable signs of apoptosis and large areas of necrosis over a 10 week examination period. By combining the in vitro and in vivo experiments, the results have suggested that this dual drug delivery system could effectively inhibit inflammation in the initial stage and prevent tumor recurrence for a relatively long term, and it may find application as a local implantable scaffold to treat a tissue defect after tumor resection.


Oncologists' reluctance to accrue patients onto clinical trials: an Illinois Cancer Center study.
Clinical trials are recognized as the standard of care for the cancer patient, and the randomized, controlled trial represents the most definitive method to determine the effectiveness or ineffectiveness of a cancer treatment. However, less than 3% of all eligible patients enter a clinical trial. Of the 437 physician members of the Illinois Cancer Center (ICC), 244 responded to a survey designed to determine factors that present a significant barrier to entering patients on clinical trials. Rigid protocol design was the primary deterrent to accrual, especially for medical oncologists. Surgeons, radiation oncologists, and medical oncologists differed with respect to several factors, including willingness to seek a clinical trial, tendency to treat patients off study, quality-of-life issues, and the belief that trials were too excessive in time commitment (P less than .05). Compared with hospital-based physicians, community oncologists had fewer patients on trial, were more likely to enter patients on the basis of age, and were more concerned about aspects of informed consent and the financial burden of a trial (P less than .01). One third of the physicians never pursued a clinical trial because of conflict with the priorities of individual care and excessive follow-up time. Fourteen percent indicated that they discouraged patients from participating in a clinical trial due to the risk of a patient receiving a placebo and patient follow-up requirements (P less than .05). Subgroups of physicians differ in their reluctance to accrue patients, and there are clusters of beliefs expressed by physicians concerning their clinical trial activity. Current conduct of clinical trials needs to be reassessed, and intervention studies are required to determine the best methodology to alter physician reluctance to pursue clinical trials.


Evaluating the use of locked nucleic acid capture probes in hybrid LC-MS/MS analysis of siRNA analytes.
Background: Hybrid LC-MS assays for oligonucleotides rely on capture probes to develop assays with high sensitivity and specificity. Locked nucleic acid (LNA) probes are thermodynamically superior to existing capture probes, but are not currently used for hybrid LC-MS assays. Materials & methods: Using two lipid-conjugated double-stranded siRNA compounds as model analytes, hybrid LC-MS/MS assays using LNA probes were developed. Results: The workflows demonstrated the superiority of the LNA probes, optimized sample preparation conditions to maximize analyte recovery, evaluated the need for analyte-specific internal standards, and demonstrated that advanced mass spectrometric technology can increase assay sensitivity by up to 20-fold. Conclusion: The workflow can be used in future bioanalytical studies to develop effective hybrid LC-MS/MS methods for siRNA analytes.


Vascular smooth muscle: integrator of vasoactive signals during exercise hyperemia.
The primary focus of this review is to discuss the importance of vascular smooth muscle function in mechanisms underlying exercise hyperemia in skeletal muscle. Important features of exercise hyperemia are presented and include: 1) the large magnitude of increase in blood flow, 2) the pattern of increased blood flow within and among skeletal muscle during exercise, 3) exercise hyperemia results from increases in vascular conductance produced by relaxation of vascular smooth muscle, 4) the increased blood flow is linked to the oxidative metabolism of the muscle, and 5) the increased blood flow occurs very rapidly with the initiation of exercise. A prevailing theme throughout this review is that vascular smooth muscle is a primary integrator of vasoactive signals that, in turn, regulate vascular resistance and muscle blood flow. Signal transduction pathways involved in vascular smooth muscle contraction and relaxation are discussed, with particular emphasis on the role of multiple and redundant signaling pathways for initiating a given contractile/relaxation response. We emphasize the concept that exercise hyperemia is a local phenomenon and that, during maximal exercise when most signals for vasoconstriction are still present, three primary control mechanisms are thought to regulate vasodilation and subsequent increases in vascular conductance: myogenic vascular control, metabolic vascular control, and endothelium-mediated vascular control. Experimental paradigms to test the relative importance of the predominant mechanisms thought to underlie exercise hyperemia are discussed and evaluated in light of the multiple and redundant control systems now known to contribute to control of blood flow in striated muscle tissue.


Designing safe job rotation schedules using optimization and heuristic search.
Job rotation is one method that is sometimes used to reduce exposure to strenuous materials handling; however, developing effective rotation schedules can be complex in even moderate sized facilities. The purpose of this research is to develop methods of incorporating safety criteria into scheduling algorithms to produce job rotation schedules that reduce the potential for injury. Integer programming and a genetic algorithm were used to construct job rotation schedules. Schedules were comprised of lifting tasks whose potential for causing injury was assessed with the Job Severity Index. Each method was used to design four job rotation schedules that met specified safety criteria in a working environment where the object weight, horizontal distance and repetition rate varied over time. Each rotation was assigned to a specific gender/lifting capacity group. Five versions of the integer programming search method were applied to this problem. Each version generated one job rotation schedule. The genetic algorithm model was able to create a population of 437 feasible solutions to the rotation problem. Utilizing cluster analysis, a rule set was derived from the genetic algorithm generated solutions. These rules provided guidelines for designing safe job rotation schedules without the use of a computer. The advantages and limitations of these approaches in developing administrative controls for the prevention of back injury are discussed.


Whole-body interdiction of lengthening of telomeres: a proposal for cancer prevention.
The intrinsic genetic instability of cancer cells makes age-related cancers more difficult to postpone or treat than any other age-related diseases. Any treatment that a cancer can resist by activating or inactivating specific genes is unlikely to succeed over the long term, because pre-existing cancer cells with the necessary gene expression pattern will withstand the therapy and proliferate. "Whole-body Interdiction of Lengthening of Telomeres" (WILT) is a proposal to pre-empt this problem by deleting from as many of our cells as possible the genes needed for telomere elongation. Cancers lacking these genes can never reach a life-threatening stage by altering gene expression, only by acquiring new genes, which is far more unlikely. Continuously-renewing tissues can be maintained by periodic reseeding with telomere elongation-incompetent stem cells that have had their telomeres lengthened in vitro with exogenous telomerase. Here, I describe why WILT might prove to be an exceptionally powerful anti-cancer modality.


Scientific status summary.
The Institute of Food Technologists has issued this Scientific Status Summary to provide readers with a tutorial on biofilms, their purposeful mechanism of interaction (quorum sensing), and recent findings on how to inhibit their formation.


Forty years of National Poison Data System Annual Reports.
The 40th National Poison Data System Annual Report from America's Poison Centers is published in this issue of Clinical Toxicology. This Commentary will focus on changes that have occurred over the years and emerging trends while highlighting some of the strengths and weaknesses of the system. The National Poison Data System now receives poison exposure data from all 50 states and territories of the United States, representing all levels of care and medical outcomes, and is collected in near real-time. The 2022 report is double in size from the 1983 report and shows changes in the distribution of exposure substances and at least a doubling in the percentage of calls from a health care facility, intentional exposures, and fatalities. There was also a more modest increase in cases managed in a health care facility, and a 40% drop in the percentage of cases managed in those under age 6. While total reported poison center cases have been decreasing since 2008, cases with more serious medical outcomes have steadily increased. To address the decline in the total number of less serious cases, an online tool was initiated for consumers to obtain simple individualized poisoning recommendations as an alternative to directly contacting a poison center. Fentanyl exposures reported to the National Poison Data System have increased rapidly since 2019, driven by non-prescription fentanyl. Reporting of this was facilitated by the granularity of the generic coding schema and the ability to develop new generic codes rapidly, unlike some other national data sets. Total fentanyl exposures and those with more serious outcomes show a good correlation with national data on fentanyl deaths, demonstrating the ability of the National Poison Data System to function as a surveillance tool. Fentanyl concentrations reported to the National Poison Data System Fatality module showed antemortem concentrations to be slightly lower than postmortem concentrations, little difference between postmortem peripheral and central concentrations, and single substance concentrations slightly higher than cases with multiple substances. Strengths include the breadth of the database, granularity, contemporaneous data collection, near-real-time data submission allowing for automated toxicosurveillance activities, ability to rapidly activate emergency product codes for emerging situations, a robust web-based query tool, mapping of generic codes and clinical effects to other data dictionaries, and review by toxicologists of fatalities to assign a relative contribution. Weaknesses include voluntarily reported, non-verified exposures, interpreting reported medical outcomes (non-toxic and fatalities), lag-time in collecting data on new therapeutic modalities, and mortality data varying from that reported by other sources. Poison center data gives a robust, reproducible image of the populations that utilize poison center resources and seems to mirror trends noted from other data sources. This supports the continued need for poison centers in the information age to support the management of patients potentially exposed to poisons and those who are more severely poisoned.


Adult patient with anomalous origin of anterior interventricular artery at main pulmonary artery.
From all congenital cardiopathies, anomalous origin of anterior interventricular artery occurs once per 300,0000 live births, reporting high mortality in the first year after birth. However, if good collateral circulation is available for the artery related to the abnormality, the patient may be kept asymptomatic until mature age. This is the report on a rare case of a 43-year-old patient who was oligosymptomatic, with normal ventricular function in this pathology presentation. The patient was submitted to surgical treatment with no extracorporeal circulation.


Paraoxonase polymorphism and its effect on male reproductive outcomes among Chinese pesticide factory workers.
Serum paraoxonase has been associated with the metabolism of organophosphate pesticides in humans. Molecular analysis of the human paraoxonase gene (PON1) has revealed that Arg192 homozygotes have a greater detoxifying capability than Gln192 homozygotes. We examined the effects of PON1 genotypes on male reproductive outcomes and its interaction with exposure to organophosphate pesticides. We studied 60 Chinese pesticide-factory workers and 89 textile-factory workers who were unexposed to pesticides. The respective allele frequencies of Arg192 and Gln192 were 0.62 and 0.38. Pesticide exposure among 36 exposed subjects and 12 unexposed subjects, regardless of gender, was assessed by personal measurement of pesticide residues over an entire 8-hr shift and measurement of urinary p-nitrophenol level over a 24-hr period. We analyzed semen and hormone data collected from male subjects. When the three PON1 genotypes were analyzed separately, a gene dose effect was not detected. We used the unexposed Arg192 homo/heterozygotes as the reference group, and re-analyzed the data. Exposed Arg192 homo/heterozygotes had significantly lower sperm count (chi 2 = 9.01, P < 0.01) and lower percentage of sperm with normal morphology (chi 2 = 4.18, P < 0.05) than the reference group. Both unexposed Gln192 homozygotes (chi 2 = 4.90, P < 0.05) and exposed Arg192 homo/heterozygotes (chi 2 = 10.00, P < 0.01) showed significantly lower sperm concentrations than the reference group. In addition, exposed Arg192 homo/heterozygotes had significantly higher serum LH levels (chi 2 = 7.94, P < 0.01) than the reference group. Because of a small sample size, our findings are highly preliminary. Nevertheless, it calls for further investigation of the interaction between the PON1 genotype and organophosphate pesticide exposure on male reproductive outcomes.


Imaging Characteristics of Nephrogenic Rests Versus Small Wilms Tumors: A Report From the Children's Oncology Group Study AREN03B2.
OBJECTIVE. Distinguishing nephrogenic rests from small Wilms tumors can be challenging. This retrospective study was performed to determine if imaging characteristics can be used to distinguish nephrogenic rests from Wilms tumors. MATERIALS AND METHODS. All cases of pathologically confirmed nephrogenic rests and Wilms tumors smaller than 5 cm in maximum dimension on imaging in patients younger than 5 years old were identified from the Children's Oncology Group AREN03B2 study (July 2006-August 2016). Exclusion criteria were chemotherapy before pathologic evaluation or more than 30 days between imaging and surgery; in addition, patients with nephrogenic rests occurring within or juxtaposed to a Wilms tumor and patients with diffuse hyperplastic perilobar nephroblastomatosis were excluded. Two radiologists who were blinded to pathology results assessed all lesions. The two-sample t test was used for continuous variables, and the Fisher exact test was used for categoric variables. ROC analysis was performed to determine the optimal size cutoff for distinguishing between nephrogenic rests and Wilms tumors. RESULTS. Thirty-one pathologically confirmed rests (20 perilobar, 11 intralobar) and 26 Wilms tumors smaller than 5 cm met the eligibility criteria for study inclusion. The median diameter of the nephrogenic rests was 1.3 cm (range, 0.7-3.4 cm) and the median diameter of the Wilms tumor was 3.2 cm (range, 1.8-4.9 cm) (p < 0.001). Imaging findings supportive of Wilms tumors were spherical (p < 0.001) and exophytic (p < 0.001) lesions. Perilobar rests (17/20) were more likely to be homogeneous than intralobar rests (3/11) or Wilms tumor (3/26) (p < 0.001). ROC analysis showed that the optimal size cutoff for distinguishing between nephrogenic rests and Wilms tumors was 1.75 cm. CONCLUSION. In children younger than 5 years old, the diagnosis of a Wilms tumor should be favored over a nephrogenic rest when a renal mass is spherical, exophytic, or larger than 1.75 cm. Homogeneity favors the diagnosis of perilobar nephrogenic rests, whereas intralobar rests and Wilms tumors are more likely to be inhomogeneous.


"Built on Respect and Good Honest Communication:" A Study of Partnerships Between Mental Health Providers and Community Corrections.
The prevailing approach to managing persons with criminal histories involves community supervision professionals like probation and parole officers partnering with other mental health providers to address clients' needs. The relationships between individual professionals are seldom researched, though, and the current study aims to address this deficit in the empirical literature. This study utilized interviews about professionals' perceptions of their work experiences, analyzed open-endedly to identify major themes. Mental health providers' themes included appreciation and process of collaboration, individual characteristics and roles, characteristics of collaboration, elements of interprofessional relationships, and involvement of the courts. Community supervision professionals discussed issues pertaining to appreciation and process of collaboration, individual characteristics and roles, when conflict occurs, and the lack of basic knowledge about other professionals. Second, these partnerships were examined in light of interprofessional healthcare competencies. Themes identified here resembled healthcare values and ethics competencies and roles and responsibilities competencies; healthcare competencies regarding interprofessional communication and teamwork showed partial congruence with the current themes. Overall, interprofessional collaboration is valued. This research highlights the strengths of this type of interprofessional collaboration and offers suggestions for improving the efficacy of collaboration.


Zoledronic acid prevents osteopenia and increases bone strength in a rabbit model of distraction osteogenesis.
Prolonged healing times and stress-shielding osteopenia remain problematic in distraction osteogenesis. In this study of 30 rabbits, zoledronic acid increased regenerate volume, mineralization, and tibial strength and prevented osteopenia over a 6-week period. Translation to the clinical setting, if safe, could improve outcomes in distraction osteogenesis in children. Because the external fixators for limb lengthening and reconstruction are designed to control the positions of bone fragments accurately, they also produce stress-shielding effects on the forming regenerate and surrounding bone. Osteopenia, leading to refracture and limitations on rehabilitation, are common consequences, potentially increasing morbidity and detracting from final clinical outcome. We examined the effect of zoledronic acid on distraction osteogenesis in 42 immature male NZW rabbits. The model chosen results in reliable regenerate formation and stress-shielding osteopenia. Fourteen animals received either Saline, zoledronic acid 0.1 mg/kg at surgery (ZOL), or another dose 2 weeks postoperatively (Redosed ZOL). Rabbits underwent DXA for bone mineral content and bone mineral density in regenerate and surrounding segments of operated and contralateral tibias. After death at 6 weeks, 30 pairs of tibias underwent quantitative computerized tomography (QCT) and four-point bend testing, and 12 were examined by histomorphometry. The study was powered at 0.8 to show differences of 1.3 SDs for mineral and mechanical parameters. Osteopenia observed in tibias of the Saline group was absent in ZOL and Redosed ZOL tibias, the latter exhibiting higher bone mineral density and bone mineral content over contralateral regions (p < 0.01). Regenerate bone mineral content was higher in ZOL and Redosed ZOL versus Saline groups at 4 and 6 weeks (p < 0.01). Cross-sectional area was 49% and 59% greater at 6 weeks in ZOL and Redosed ZOL regenerates compared with the Saline group (p < 0.01). ZOL and Redosed ZOL tibias were 29% and 89% stronger by four-point bending than the Saline group (p < 0.01). Histomorphometry in the regenerate of ZOL and Redosed ZOL groups revealed higher trabecular bone volume and trabecular number compared with the Saline group (p < 0.001). Zoledronic acid administration led to significantly greater bone area, mineral content, strength, and trabecular number with reduced stress-shielding osteopenia in this model of distraction osteogenesis. These data suggest that intraoperative and postoperative zoledronic acid administration could improve outcomes in children undergoing limb lengthening.


Moxibustion for treating knee osteoarthritis: study protocol of a multicentre randomised controlled trial.
The treatment of knee osteoarthritis, which is a major cause of disability among the elderly, is typically selected from multidisciplinary options, including complementary and alternative medicine. Moxibustion has been used in the treatment of knee osteoarthritis in Korea to reduce pain and improve physical activity. However, there is no sufficient evidence of its effectiveness, and it cannot therefore be widely recommended for treating knee osteoarthritis. We designed a randomised controlled clinical trial to evaluate the effectiveness, safety, cost-effectiveness, and qualitative characteristics of moxibustion treatment of knee osteoarthritis compared to usual care. This is a protocol for a multicentre, pragmatic, randomised, assessor-blinded, controlled, parallel-group study. A total of 212 participants will be assigned to the moxibustion group (n = 106) and the usual care group (n = 106) at 4 clinical research centres. The participants assigned to the moxibustion group will receive moxibustion treatment of the affected knee(s) at 6 standard acupuncture points (ST36, ST35, ST34, SP9, Ex-LE04, and SP10) 3 times per week for 4 weeks (a total of 12 sessions). Participants in the usual care group will not receive moxibustion treatment during the study period. Follow-up will be performed on the 5th and 13th weeks after random allocation. Both groups will be allowed to use any type of treatment, including surgery, conventional medication, physical treatment, acupuncture, herbal medicine, over-the-counter drugs, and other active treatments. Educational material that explains knee osteoarthritis, the current management options, and self-exercise will be provided to each group. The global scale of the Korean Western Ontario and McMaster Osteoarthritis Index (K-WOMAC) will be the primary outcome measurement used in this study. Other subscales (pain, stiffness, and function) of the K-WOMAC, the Short-Form 36v2 Health Survey, the Beck Depression Inventory, the Physical Function test, Patient Global Assessment, and the Pain Numerical Rating Scale will be used as outcome variables to evaluate the effectiveness of moxibustion. Safety will be assessed at every visit. In addition, an economic evaluation and a qualitative study will be conducted as a mixed-methods approach. This trial may contribute to developing evidence for the effectiveness and safety of moxibustion for treating knee osteoarthritis. KCT0000130.


Current Perspectives in the Application of Medicinal Plants Against Cancer: Novel Therapeutic Agents.
Cancer is a disease characterized by uncontrolled cell growth and proliferation. It has become a major health problem in the past decades and is now the second leading cause of death globally. Although, there are different types of treatment such as chemotherapy, immune therapy, radiation, hormone therapy and targeted therapy used against cancer, they have possible side effects and significant deficiencies. This review aims to outline the benefits of medicinal plants and plant-derived products and highlight why they should be used as novel anti-cancer therapeutics. Electronic databases, including PubMed, Scopus, ScienceDirect, Cochrane library, and MedlinePlus were searched to summarize in vitro, in vivo and clinical studies on anticancer effects of medicinal plants and their bioactive compounds up-to-date. In recent years, a number of medicinal plants have been administered to cancer patients in order to prevent and treat cancer as an alternative therapy. These plants were used because of their rich anticarcinogenic and chemoprotective potentials. In addition to these remarkable properties, these plants have less toxic anticancer, anti-tumor and anti-proliferation agents than traditional therapeutics. Nevertheless, only a small number of natural anti-tumor products including vinblastine, vincristine, podophyllotoxin, paclitaxel (Taxol) and camptothecin have been tested clinically, while vinflunine ditartrate, anhydrovinblastine, NK-611, tafluposide, paclitaxel poliglumex, combretastatins, salvicine, curcumin, indirubin, triptolide, homoharringtonine are still on trial. Consequently, more effective anticancer compounds are identified during the clinical trials; these natural products could be a key source of antitumor agents in modern anticancer therapy. It is expected that novel anticancer phytopharmaceuticals produced from medicinal plants could be effectively used in prevention and therapy for the cancers.


Routine postaccess-closure angiography to detect vascular complications following transfemoral TAVR.
Vascular complications following transfemoral TAVR are associated with increased morbidity and mortality. Measures that may mitigate this risk are important. To evaluate the utility of routine, access-vessel angiography post sheath-removal in the detection and management of complications in patients undergoing transcatheter aortic valve replacement (TAVR). This was a retrospective study of 512 consecutive patients who underwent transfemoral TAVR with routine post access-closure angiography from the radial artery. Rates of mild angiographically evident bleeding, bleeding requiring surgery/interventional-radiology, ischemia, 90-day access-site-related events, and major and minor vascular complications using Valve Academic Research Consortium 3 definitions were recorded. Of 512 patients, digital subtraction angiography (DSA) was undertaken via the radial artery in 467 patients (91%). In the remaining patients (9%) DSA was either not attempted, due to concerns regarding kidney disease and contrast volume, or failed due to anatomical factors (aortic tortuosity/calcification). Significant chronic kidney disease was present at baseline in 72.4% of this cohort (stages III-IV or dialysis). Ninety-four percent of cases underwent TAVR using a balloon-expandable platform. Mild iliofemoral extravasation was observed in 7.7% of the DSA cases. These cases were managed by manual compression with none requiring any vascular intervention subsequently. Valve Academic Research Consortium 3 major and minor access-site-related complications were observed in 0.4% and 12.2%, respectively. Access-site-related bleeding and ischemic events requiring interventional-radiology or vascular-surgery were observed in 0.9% and 1.7% of the DSA cases, respectively. No new renal replacement therapy was needed in any of the DSA cases. Discharge to 90-day access-related complications was 0.8%. Routine post access-closure angiography is feasible via the radial artery in patients undergoing transfemoral TAVR and appears safe. It facilitates early identification of complications and mitigates risk by enabling prompt action to be taken. Larger studies are needed to confirm these findings.


Associations among maternal characteristics, labour interventions, delivery mode and maternal antenatal complications in a NSW large rural town, and comparison to NSW state data.
To determine associations between maternal characteristics, labour interventions, delivery mode and maternal antenatal complications in a NSW rural hospital, and compare these to state data. Data (maternal characteristics, labour type, delivery mode and maternal antenatal complications) pertaining to women who had delivered a singleton birth at Griffith Base hospital were analysed. Retrospective data analysis of Griffith Base Hospital 'eMaternity' database SETTING: Single large rural town in NSW. Women who delivered a singleton birth between July 2018 and June 2019 inclusive at Griffith Base Hospital. 1. Comparison of maternal characteristics of age, BMI, gravida, parity and gestation data between labour type (spontaneous, augmented, induced and planned caesarean section) and delivery modes (vaginal, instrumental vaginal and caesarean section). 2. Associations between labour type, augmentation or induction method, delivery mode and maternal antenatal complications. 3. Multiple regression analysis for influence of age, BMI, parity and labour type on emergency caesarean section outcome. 4. Comparison of maternal, labour and delivery data with NSW state data. Among 457 women, there were higher rates of obesity and spontaneous labour, lower rates of planned caesarean section and augmented labour, and similar rates of induction of labour and emergency caesarean section, compared with NSW. Emergency caesarean section was significantly associated with older age (β = 0.163), and labour augmentation (β = 0.114) and induction (β = 0.169). Labour augmentation with synthetic oxytocin, and induction with balloon catheter, were associated with the highest rates of emergency caesarean section. This large rural town had fewer labour and delivery medical interventions compared with NSW overall. Augmentation and induction of labour contribute to increasing caesarean section rates: directly via associations with emergency caesarean section, and indirectly because previous caesarean section was the most common reason for elective caesarean section.


Biclonal myelodysplastic syndrome involving six chromosomes and monoallelic loss of RB1 - A rare case.
Myelodysplastic syndrome (MDS) represents a group of clonal hematological disorders characterized by progressive cytopenia, and reflects to defects in erythroid, myeloid and megakaryocytic maturation. MDS is more frequently observed in older aged patients with cytogenetic abnormalities like monosomy of chromosome(s) 5 and/or 7. In 50% of de novo MDS cases, chromosomal aberrations are found and rearrangements involving the retinoblastoma (RB1) gene in 13q14 are found. Here, we are presenting a case report of a rare biclonal MDS with a karyotype of 45, XY,-4, der(6)t(4;6)(p15.1;p21.3), der(8)t(4;8)(q31.2;q22), t(13;16)(q21.3;p11.2)11/45, XY, der(7)t(7;13)(p22.2~22.3;q21.3),-13 9. The patient was diagnosed according to WHO classification as refractory anemia with excess of blasts (RAEB-II).Immunophenotyping was positive for CD11b, CD11c, CD10, CD13, CD15, CD16 and CD33. We report, a novel and cytogenetically rare case of a biclonal MDS with complex chromosomal aberrations and deletion of RB1-gene in both clones. These findings are associated with a poor prognosis as the patient died 3 months after diagnosis.


Evaluation of lasalocid in salt against ovine Coccidia.
Lasalocid at a level of 0.75% in free-choice granulated salt was highly efficacious against naturally acquired coccidia infections in lambs maintained in semiconfinement. Treated lambs consumed 18 g of medicated salt/day, reflecting an average daily lasalocid dosage of 4.3 mg/kg of body weight. Treated lambs gained 5.7 kg more than did untreated lambs (P less than 0.01) during the 84-day experiment. Clinical signs of coccidiosis were not observed in treated or untreated lambs.


Ingestion of Sodium Bicarbonate (NaHCO3) Following a Fatiguing Bout of Exercise Accelerates Postexercise Acid-Base Balance Recovery and Improves Subsequent High-Intensity Cycling Time to Exhaustion.
This study evaluated the ingestion of sodium bicarbonate (NaHCO3) on postexercise acid-base balance recovery kinetics and subsequent high-intensity cycling time to exhaustion. In a counterbalanced, crossover design, nine healthy and active males (age: 23 ± 2 years, height: 179 ± 5 cm, body mass: 74 ± 9 kg, peak mean minute power (Wpeak) 256 ± 45 W, peak oxygen uptake (V̇O2peak) 46 ± 8 ml.kg-1.min-1) performed a graded incremental exercise test, two familiarization and two experimental trials. Experimental trials consisted of cycling to volitional exhaustion (TLIM1) at 100% WPEAK on two occasions (TLIM1 and TLIM2) interspersed by a 90 min passive recovery period. Using a double-blind approach, 30 min into a 90 min recovery period participants ingested either 0.3 g.kg-1 body mass sodium bicarbonate (NaHCO3) or a placebo (PLA) containing 0.1 g.kg-1 body mass sodium chloride (NaCl) mixed with 4 ml.kg-1 tap water and 1 ml.kg-1 orange squash. The mean differences between TLIM2 and TLIM1 was larger for PLA compared with NaHCO3 (-53 ± 53 vs. -20 ± 48 s; p = .008, d = 0.7, CI =-0.3, 1.6), indicating superior subsequent exercise time to exhaustion following NaHCO3. Blood lactate [Bla-] was similar between treatments post TLIM1, but greater for NaHCO3 post TLIM2 and 5 min post TLIM2. Ingestion of NaHCO3 induced marked increases (p < .01) in both blood pH (+0.07 ± 0.02, d = 2.6, CI = 1.2, 3.7) and bicarbonate ion concentration [HCO3-] (+6.8 ± 1.6 mmo.l-1, d = 3.4, CI = 1.8, 4.7) compared with the PLA treatment, before TLIM2. It is likely both the acceleration of recovery, and the marked increases of acid-base after TLIM1 contributed to greater TLIM2 performance compared with the PLA condition.


[Antioncogene p53 and apoptosis response: new hypotheses on the molecular bases of tumor resistance to radiotherapy].
Recent experimental data indicate that in normal or neoplastic cells, DNA damage induced by ionizing radiation may work as a signal to trigger apoptosis, a characteristic suicide program inherent to multicellular organisms. The morphological signatures of apoptosis are cell shrinkage, nuclear condensation and genomic fragmentation down to the size of individual nucleosome units (apoptotic bodies). A number of genes have been identified as involved regulators of apoptosis. The tumor suppressor gene p53 has emerged as a main modulator and inducer of apoptosis. Cells containing wild-type p53 function undergo G1 arrest following DNA damage by ionizing radiations. DNA repair processes are, then, activated before the damaged genome can be replicated, with resultant genetic instability. If DNA repair fails, the wild p53 gene may trigger apoptosis and the cell with persisting DNA damage, dies. Cells with loss of functional p53 gene by either gene mutation or deletion appear unable to undergo radiation-induced apoptosis. This evidence may underlie the increased likelihood for p53 deficient cells to develop oncogenetic processes after irradiation exposure. According to p53-triggering apoptosis patterns, cancer cells with a normal p53 gene might be more sensitive to radiotherapy and less proliferating than tumor cells with p53 deficient function. If these data are confirmed, tumors harboring p53 gene mutation might be managed best with particularly aggressive or experimental treatment protocols. Conversely, a higher therapeutic index between malignant and normal cells might be obtained in tumors with fast activation of p53 dependent apoptotic response. Before any definitive conclusions are drawn, further knowledge of the different genes which are known to be involved in the regulation of apoptosis, is warranted.


Determination of seasonality in southern hairy-nosed wombats (Lasiorhinus latifrons) by analysis of fecal androgens.
Little is known about the reproductive biology of Australia's critically endangered northern hairy-nosed wombat (Lasiorhinus krefftii), largely due to its cryptic nature and the difficulty in accessing the small remaining population of about 70 animals. Using the noninvasive technique of fecal steroid analysis, we have examined the endocrinology of the more common yet closely related southern hairy-nosed wombat (Lasiorhinus latifrons). The aims of this study were to 1) develop and validate fecal androgen analysis in this species, 2) examine and compare seasonal differences in fecal and plasma androgens in male wombats, and 3) correlate seasonal differences in androgens with changes in male accessory glands (prostate and bulbourethral gland). Fecal androgens were extracted in ether; concentrated; separated by HPLC into testosterone (T), dihydrotestosterone (DHT), and 5 alpha-androstane-3 alpha,17 beta-diol (Adiol) fractions; and quantitated by RIA. The concentrations of androgens in fecal pellets from 14 wild southern hairy-nosed wombats as determined by RIA varied over the range 6.6-25.0 ng/g dry weight for T, 4.0-24.2 ng/g dry weight for DHT, and 0-34.8 ng/g dry weight for Adiol. For each androgen, a highly significant linear correlation was observed between plasma and fecal concentrations. When individuals were grouped into either breeding season (pellets collected between August-November) or nonbreeding season (collected between February-April), significant (P < 0.05) differences between seasons were observed for both plasma and fecal T, plasma DHT, and fecal Adiol. For all androgens, the mean fecal and plasma concentrations were higher during the breeding season than the nonbreeding season. A significant (P < 0.001) correlation was observed between fecal T and prostate weight, while DHT and Adiol correlations were nonsignificant. Significant correlations were observed, however, between all three fecal androgens and bulbourethral gland weight. These studies demonstrate that fecal T is a valid indicator of reproductive status in the male southern hairy-nosed wombat, with significant correlations observed between fecal T, plasma T, and prostate and bulbourethral gland weights. These findings have important implications for the study of the reproductive endocrinology of the critically endangered northern hairy-nosed wombat.


Quantum dot nanocrystals having guanosine imprinted nanoshell for DNA recognition.
Molecular imprinted polymers (MIPs) as a recognition element for sensors are increasingly of interest and MIP nanoparticles have started to appear in the literature. In this study, we have proposed a novel thiol ligand-capping method with polymerizable methacryloylamido-cysteine (MAC) attached to CdS quantum dots (QDs), reminiscent of a self-assembled monolayer and have reconstructed surface shell by synthetic host polymers based on molecular imprinting method for DNA recognition. In this method, methacryloylamidohistidine-platinium (MAH-Pt(II)) is used as a new metal-chelating monomer via metal coordination-chelation interactions and guanosine templates of DNA. Nanoshell sensors with guanosine templates give a cavity that is selective for guanosine and its analogues. The guanosine can simultaneously chelate to Pt(II) metal ion and fit into the shape-selective cavity. Thus, the interaction between Pt(II) ion and free coordination spheres has an effect on the binding ability of the CdS QD nanosensor. The binding affinity of the guanosine imprinted nanocrystals has investigated by using the Langmuir and Scatchard methods, and experiments have shown the shape-selective cavity formation with O6 and N7 of a guanosine nucleotide (K(a) = 4.841x10(6) mol L(-1)) and a free guanine base (K(a) = 0.894x10(6) mol L(-1)). Additionally, the guanosine template of the nanocrystals is more favored for single stranded DNA compared to double stranded DNA.


Interplay between intercellular signaling and cell movement in development.
Cell movement and local intercellular signaling are crucial components of morphogenesis during animal development. Intercellular signaling regulates the collective movement of a cell population via direct cell-cell contact. Cell movement, conversely, can influence local intercellular signaling by rearranging neighboring cells. Here, we first discuss theoretical models that address how intercellular signaling regulates collective cell movement during development. Examples include neural crest cell migration, convergent extension, and cell movement during vertebrate axis elongation. Second, we review theoretical studies on how cell movement may affect intercellular signaling, using the segmentation clock in zebrafish as an example. We propose that interplay between cell movement and intercellular signaling must be considered when studying morphogenesis in embryonic development.


Low-oxygen culture conditions extend the multipotent properties of human retinal progenitor cells.
Development of an effective cell-based therapy is highly dependent upon having a reproducible cell source suitable for transplantation. One potential source, isolated from the developing fetal neural retina, is the human retinal progenitor cell (hRPC). One limiting factor for the use of hRPCs is their in vitro expansion limit. As such, the aim of this study was to determine whether culturing hRPCs under 3% O2 would support their proliferative capacity while maintaining multipotency. To determine the effect of low oxygen on the ability of hRPCs to self-renew, rates of proliferation and apoptosis, telomerase activity, and expression of proliferative, stemness, and differentiation markers were assessed for hRPCs cultured in 3% and 20% oxygen conditions. Culture under 3% oxygen increases the proliferation rate and shifts the proliferation limit of hRPCs to greater 40 divisions. This increased capacity for proliferation is correlated with an upregulation of Ki67, CyclinD1, and telomerase activity and a decrease in p53 expression and apoptosis. Increased expression of cMyc, Klf4, Oct4, and Sox2 in 3% O₂ is correlated with stabilization of both HIF1α and HIF2α. The eye field development markers Pax6, Sox2, and Otx2 are present in hRPCs up to passage 16 in 3% O₂ . Following in vitro differentiation hRPCs expanded in the 3% O₂ were able to generate specialized retinal cells, including rods and cones. Low-oxygen culture conditions act to maintain both multipotency and self-renewal properties of hRPCs in vitro. The extended expansion limits permit the development of a clinical-grade reagent for transplantation.


Social cognitive theory mediators of physical activity in a lifestyle program for cancer survivors and carers: findings from the ENRICH randomized controlled trial.
Despite increasing numbers of cancer survivors and evidence that diet and physical activity improves the health of cancer survivors, most do not meet guidelines. Some social cognitive theory (SCT)-based interventions have increased physical activity behavior, however few have used objective physical activity measures. The Exercise and Nutrition Routine Improving Cancer Health (ENRICH) randomized controlled trial reported a significant intervention effect for the primary outcome of pedometer-assessed step counts at post-test (8-weeks) and follow-up (20-weeks). The aim of this study was to test whether the SCT constructs operationalized in the ENRICH intervention were mediators of physical activity behavior change. Randomized controlled trial with 174 cancer survivors and carers assessed at baseline, post-test (8-weeks), and follow-up (20-weeks). Participants were randomized to the ENRICH six session face-to-face healthy lifestyle program, or to a wait-list control. Hypothesized SCT mediators of physical activity behavior change (self-efficacy, behavioral goal, outcome expectations, impediments, and social expectations) were assessed using valid and reliable scales. Mediation was assessed using the Preacher and Hayes SPSS INDIRECT macro. At eight weeks, there was a significant intervention effect on behavioral goal (A = 9.12, p = 0.031) and outcome expectations (A = 0.25, p = 0.042). At 20 weeks, the intervention had a significant effect on self-efficacy (A = 0.31, p = 0.049) and behavioral goal (A = 13.15, p = 0.011). Only changes in social support were significantly associated with changes in step counts at eight weeks (B = 633.81, p = 0.023). Behavioral goal was the only SCT construct that had a significant mediating effect on step counts, and explained 22 % of the intervention effect at 20 weeks (AB = 397.9, 95 % CI 81.5-1025.5). SCT constructs had limited impact on objectively-assessed step counts in a multiple health behavior change intervention for cancer survivors and their carers. Behavioral goal measured post-intervention was a significant mediator of pedometer-assessed step counts at 3-months after intervention completion, and explained 22 % of the intervention effect. Future research should examine the separate impact of goals and planning, as well as examining mediators of behavior maintenance in physical activity interventions targeting cancer survivors. Australian and New Zealand Clinical Trials registry ANZCTRN1260901086257 .


Bronchial obstruction and exhaled nitric oxide response during exercise in cold air.
This study examines whether exhausting exercise in cold air induces bronchial obstruction and changes in exhaled [NO] and in exhaled NO output (V'NO). Thus, eight well-trained males performed two incremental exercise tests until exhaustion, followed by 5 min of recovery in temperate (22 degrees C) and cold (-10 degrees C) environments, at random. At -10 degrees C, they were dressed in warm clothes. Ventilation (V'E), oxygen consumption (V'O2), carbon dioxide production, cardiac frequency (fC), and [NO] and V'NO were measured continuously. Before and after each test, the subjects' maximal expiratory flow-volume curves and peak expiratory flow, forced expiratory volume in one second (FEV1) and forced expiratory flow at 25 (FEF25), 50 (FEF50) and 75% (FEF75) of forced vital capacity were determined. At -10 degrees C, significant decreases in FEV1 and FEF75 were observed after exercise. At rest and at the same submaximal intensity, V'O2, V'E and fC did not differ significantly. At rest and up to approximately 50% peak V'O2, [NO] and V'NO values were lower at -10 degrees C than at 22 degrees C. Thereafter, and during recovery, the V'NO response became similar at both -10 and 22 degrees C. This study confirms that considerable hyperpnoea in cold air causes a detectable airway obstruction. This airway cooling also induces an initial decrease in the exhaled NO response. Since endogenous NO-production is involved in bronchial dilation, it cannot be excluded that this lack of production may favour the appearance of airway obstruction.


Inhibition of Proinflammatory Cytokine Release by Flavones and Flavanones from the Leaves of Dracaena steudneri Engl.
The leaves of Dracaena steudneri yielded 6 new flavonoids-3,5,7-trihydroxy-6-methyl-3',4'-methylenedioxyflavone (1: ), 5,7-dihydroxy-3-methoxy-6-methyl-3',4'-methylenedioxyflavone (2: ), 3,5,7-trihydroxy-6-methoxy-3',4'-methylenedioxyflavone (3: ), (2S,3S)-3,7-dihydroxy-6-methoxy-3',4'-methylenedioxyflavanone (4: ), 4',5,7-trihydroxy-3,3',8-trimethoxy-6-methylflavone (5: ), (2R) 7-hydroxy-2',8-dimethoxyflavanone (6: )-together with 13 known congeners. Their structures were established using spectroscopic and spectrometric methods including NMR, CD, and HRMSn measurements. The compounds were evaluated for their anti-inflammatory potential through measurement of the levels of cytokines IL-1β, IL-2, GM-CSF, and TNF-α in the supernatant of human peripheral blood mononuclear cells stimulated by lipopolysaccharide. Flavones derivatives 1: -4: with a C-3'/4' methylenedioxy substituent led to a substantial increase in the production of IL-1β and GM-CSF out of 4 pro-inflammatory cytokines relative to LPS control. Quercetin derivatives 5, 11,: and 13: with a hydroxyl group at C-4' inhibited the production of IL-2, GM-CSF, and TNF-α. The presence of a C-2/C-3 double bond in 14: was pivotal to the significantly stronger (0.4 to 27.5% of LPS control) inhibitory effect compared to its dihydro derivative 8: (36.2 to 262.7% of LPS control) against all tested cytokines. It is important to note that the inhibitory activity of 14: was substantially higher than that of the standard drug used, ibuprofen.


MCAK regulates chromosome alignment but is not necessary for preventing aneuploidy in mouse oocyte meiosis I.
Errors in chromosome segregation in mammalian oocytes lead to aneuploid eggs that are developmentally compromised. In mitotic cells, mitotic centromere associated kinesin (MCAK; KIF2C) prevents chromosome segregation errors by detaching incorrect microtubule-kinetochore interactions. Here, we examine whether MCAK is involved in spindle function in mouse oocyte meiosis I, and whether MCAK is necessary to prevent chromosome segregation errors. We find that MCAK is recruited to centromeres, kinetochores and chromosome arms in mid-meiosis I, and that MCAK depletion, or inhibition using a dominant-negative construct, causes chromosome misalignment. However, the majority of oocytes complete meiosis I and the resulting eggs retain the correct number of chromosomes. Moreover, MCAK-depleted oocytes can recover from mono-orientation of homologous kinetochores in mid-meiosis I to segregate chromosomes correctly. Thus, MCAK contributes to chromosome alignment in meiosis I, but is not necessary for preventing chromosome segregation errors. Although other correction mechanisms may function in mammalian meiosis I, we speculate that late establishment of kinetochore microtubules in oocytes reduces the likelihood of incorrect microtubule-kinetochore interactions, bypassing the requirement for error correction.


Effects of saw palmetto extract on micturition reflex of rats and its autonomic receptor binding activity.
We examined the effects of saw palmetto extract (SPE) on the rat micturition reflex and on autonomic receptors in the lower urinary tract. The effect of SPE was examined on cystometrograms of anesthetized rats induced by intravesical infusion of saline or 0.1% acetic acid. SHR/NDmc-cp (cp/cp) rats received repeat oral administration of SPE and nighttime urodynamic function was determined. The autonomic receptor binding activity of SPE in the rat bladder and prostate was examined by radioligand binding assay. Intraduodenal administration of SPE (60 mg/kg) in anesthetized rat cystometry caused a significant increase in the micturition interval, micturition volume and bladder capacity during intravesical saline infusion. Also, similar administration of SPE at doses of 12 and 20 mg/kg significantly reversed the shortened micturition interval as well as the decreased micturition volume and bladder capacity due to 0.1% acetic acid infusion in a dose dependent manner. In conscious SHR/NDmc-cp (cp/cp) rats repeat oral administration of SPE (6 mg/kg daily) constantly increased the micturition interval and concomitantly decreased voiding frequency. SPE inhibited specific binding of [H]NMS ([N-methyl-H]scopolamine methyl chloride) (bladder) and [H]prazosin (prostate) with IC50 values of 46.1 and 183 microg/ml, respectively. SPE significantly alleviates urodynamic symptoms in hyperactive rat bladders by increasing bladder capacity and subsequently prolonging the micturition interval. Our data may support the clinical efficacy of SPE for the treatment of lower urinary tract symptoms.


Enhancement of neutrophil autophagy by an IVIG preparation against multidrug-resistant bacteria as well as drug-sensitive strains.
Autophagy occurs in human neutrophils after the phagocytosis of multidrug-resistant bacteria and drug-sensitive strains, including Escherichia coli and Pseudomonas aeruginosa. The present study detected autophagy by immunoblot analysis of LC3B conversion, by confocal scanning microscopic examination of LC3B aggregate formation and by transmission electron microscopic examination of bacteria-containing autophagosomes. Patients with severe bacterial infections are often treated with IVIG alongside antimicrobial agents. Here, we showed that IVIG induced neutrophil-mediated phagocytosis of multidrug-resistant strains. Compared with untreated neutrophils, neutrophils exposed to IVIG showed increased levels of bacterial cell killing, phagocytosis, O(2)(-) release, MPO release, and NET formation. IVIG also increased autophagy in these cells. Inhibiting the late phase of autophagy (fusion of lysosomes with autophagosomes) with bafilomycin A1-reduced, neutrophil-mediated bactericidal activity. These findings indicate that autophagy plays a critical role in the bactericidal activity mediated by human neutrophils. Furthermore, the autophagosomes within the neutrophils contained bacteria only and their organelles only, or both bacteria and their organelles, a previously undocumented observation. Taken together, these results suggest that the contents of neutrophil autophagosomes may be derived from specific autophagic systems, which provide the neutrophil with an advantage. Thus, IVIG promotes the neutrophil-mediated killing of multidrug-resistant bacteria as well as drug-sensitive strains.


Human dendritic cell maturation by adenovirus transduction enhances tumor antigen-specific T-cell responses.
Dendritic cells (DCs) have been shown to require a degree of maturation to stimulate antigen-specific, type 1 cytotoxic T lymphocytes in numerous murine models. Limited data in humans suggest that immature DCs (DC) can induce tolerance, yet a variety of nonmatured DC used clinically have induced antigen-specific type 1 T cells in vivo to various tumor-associated antigens. Use of adenovirus to engineer DCs is an efficient method for delivery of entire genes to DC, but the data on the biologic effects of viral transduction are contradictory. The authors demonstrate that DCs transduced with adenovirus (AdV) clearly become more mature by the phenotypic criterion of upregulation of CD83 and downregulation of CD14. Transduced DCs also decrease production of IL-10, and a subset of transduced DCs produce increased levels of IL-12 p70. This level of maturation is superior to that achieved by treatment of these cells with tumor necrosis factor-alpha or interferon-alpha but less pronounced than with CD40L trimer or CD40L + interferon-gamma. Maturation by AdV transduction alone leads to efficient stimulation of antigen-specific T cells from both healthy donors and patients with advanced cancer using two defined human tumor-associated antigens, MART-1 and AFP. Given the pivotal role of DCs in immune activation, it is important to understand the direct biologic effects of AdV on DCs, as well as the impact these biologic changes have on the stimulation of antigen-specific T cells. This study has important implications for the design of DC-based clinical trials.


Dietary omega 3 fatty acids and the developing brain.
The omega-3 fatty acids are essential dietary nutrients and one of their important roles is providing the fatty acid with 22 carbons and 6 double bonds known as docosahexaenoic acid (DHA) for nervous tissue growth and function. Inadequate intakes of omega-3 fatty acids decrease DHA and increase omega-6 fatty acids in the brain. Decreased DHA in the developing brain leads to deficits in neurogenesis, neurotransmitter metabolism, and altered learning and visual function in animals. Western diets are low in omega-3 fatty acids, including the 18 carbon omega-3 fatty acid alpha linolenic acid found mainly in plant oils, and DHA, which is found mainly in fish. The DHA status of the newborn and breast-fed infant depends on the maternal intake of DHA and varies widely. Epidemiological studies have linked low maternal DHA to increased risk of poor child neural development. Intervention studies have shown improving maternal DHA nutrition decreases the risk of poor infant and child visual and neural development. Thus, sufficient evidence is available to conclude that maternal fatty acid nutrition is important to DHA transfer to the infant before and after birth, with short and long-term implications for neural function. However, genetic variation in genes encoding fatty acid desaturases also influence essential fatty acid metabolism, and may increase requirements in some individuals. Consideration of omega-3 fatty acid to include brain development, optimizing omega-3 and omega-6 fatty acids in gestation and lactation, and in fatty acid nutrition support for intravenous and formula-fed neonates is important.


Copper egress is induced by PMA in human THP-1 monocytic cell line.
Copper egress is an essential regulator of the kinetics of cellular copper and is primarily regulated by ATP7A, a copper-transporting P-type ATPase. However, little is known under which physiological condition copper egress is induced and its molecular consequence. In current manuscript, using THP-1 cells, a human monocytic cell line, we found that ATP7A expression was increased in cells exposed to phorbol-12-myristate-13-acetate (PMA), a potent inducer of neovascularization and cancer. Inductively coupled plasma mass spectrometry revealed that PMA also induced copper egress. Inhibition of ATP7A expression using small interfering RNA abrogated PMA induced copper egress. PMA treatment in THP-1 cells resulted in increased expression of matrix metalloproteinase (MMP) 9 and vascular endothelial growth factor receptor 1 (VEGFR1), whereas inhibition of ATP7A resulted in suppression of PMA-induced expression of VEGFR1, but not MMP9. Finally, addition of exogenous copper into the conditioned medium did not change VEGFR1 expression in THP-1 cells. Collectively, we demonstrate that PMA induces copper egress in THP-1 cells, which is regulated by ATP7A, and ATP7A regulates VEGFR1 expression. Considering the involvement of copper in neovascularization, our current finding provides the potential evidence to interpret the molecular mechanism.


Cocaine Induces Inflammatory Gut Milieu by Compromising the Mucosal Barrier Integrity and Altering the Gut Microbiota Colonization.
Cocaine use disorder (CUD), a major health crisis, has traditionally been considered a complication of the CNS; however, it is also closely associated with malnourishment and deteriorating gut health. In light of emerging studies on the potential role of gut microbiota in neurological disorders, we sought to understand the causal association between CUD and gut dysbiosis. Using a comprehensive approach, we confirmed that cocaine administration in mice resulted in alterations of the gut microbiota. Furthermore, cocaine-mediated gut dysbiosis was associated with upregulation of proinflammatory mediators including NF-κB and IL-1β. In vivo and in vitro analyses confirmed that cocaine altered gut-barrier composition of the tight junction proteins while also impairing epithelial permeability by potentially involving the MAPK/ERK1/2 signaling. Taken together, our findings unravel a causal link between CUD, gut-barrier dysfunction and dysbiosis and set a stage for future development of supplemental strategies for the management of CUD-associated gut complications.


A critical assessment of comparative molecular modeling of tertiary structures of proteins.
In spite of the tremendous increase in the rate at which protein structures are being determined, there is still an enormous gap between the numbers of known DNA-derived sequences and the numbers of three-dimensional structures. In order to shed light on the biological functions of the molecules, researchers often resort to comparative molecular modeling. Earlier work has shown that when the sequence alignment is in error, then the comparative model is guaranteed to be wrong. In addition, loops, the sites of insertions and deletions in families of homologous proteins, are exceedingly difficult to model. Thus, many of the current problems in comparative molecular modeling are minor versions of the global protein folding problem. In order to assess objectively the current state of comparative molecular modeling, 13 groups submitted blind predictions of seven different proteins of undisclosed tertiary structure. This assessment shows that where sequence identity between the target and the template structure is high (> 70%), comparative molecular modeling is highly successful. On the other hand, automated modeling techniques and sophisticated energy minimization methods fail to improve upon the starting structures when the sequence identity is low (approximately 30%). Based on these results it appears that insertions and deletions are still major problems. Successfully deducing the correct sequence alignment when the local similarity is low is still difficult. We suggest some minimal testing of submitted coordinates that should be required of authors before papers on comparative molecular modeling are accepted for publication in journals.


ICC-CLASS: isotopically-coded cleavable crosslinking analysis software suite.
Successful application of crosslinking combined with mass spectrometry for studying proteins and protein complexes requires specifically-designed crosslinking reagents, experimental techniques, and data analysis software. Using isotopically-coded ("heavy and light") versions of the crosslinker and cleavable crosslinking reagents is analytically advantageous for mass spectrometric applications and provides a "handle" that can be used to distinguish crosslinked peptides of different types, and to increase the confidence of the identification of the crosslinks. Here, we describe a program suite designed for the analysis of mass spectrometric data obtained with isotopically-coded cleavable crosslinkers. The suite contains three programs called: DX, DXDX, and DXMSMS. DX searches the mass spectra for the presence of ion signal doublets resulting from the light and heavy isotopic forms of the isotopically-coded crosslinking reagent used. DXDX searches for possible mass matches between cleaved and uncleaved isotopically-coded crosslinks based on the established chemistry of the cleavage reaction for a given crosslinking reagent. DXMSMS assigns the crosslinks to the known protein sequences, based on the isotopically-coded and un-coded MS/MS fragmentation data of uncleaved and cleaved peptide crosslinks. The combination of these three programs, which are tailored to the analytical features of the specific isotopically-coded cleavable crosslinking reagents used, represents a powerful software tool for automated high-accuracy peptide crosslink identification. See: http://www.creativemolecules.com/CM_Software.htm.


Iridium-Catalyzed Asymmetric Hydrogenation of Unsaturated Carboxylic Acids.
Chiral carboxylic acid moieties are widely found in pharmaceuticals, agrochemicals, flavors, fragrances, and health supplements. Although they can be synthesized straightforwardly by transition-metal-catalyzed enantioselective hydrogenation of unsaturated carboxylic acids, because the existing chiral catalysts have various disadvantages, the development of new chiral catalysts with high activity and enantioselectivity is an important, long-standing challenge. Ruthenium complexes with chiral diphosphine ligands and rhodium complexes with chiral monodentate or bidentate phosphorus ligands have been the predominant catalysts for asymmetric hydrogenation of unsaturated acids. However, the efficiency of these catalysts is highly substrate-dependent, and most of the reported catalysts require a high loading, high hydrogen pressure, or long reaction time for satisfactory results. Our recent studies have revealed that chiral iridium complexes with chiral spiro-phosphine-oxazoline ligands and chiral spiro-phosphine-benzylamine ligands exhibit excellent activity and enantioselectivity in the hydrogenation of α,β-unsaturated carboxylic acids, including α,β-disubstituted acrylic acids, trisubstituted acrylic acids, α-substituted acrylic acids, and heterocyclic α,β-unsaturated acids. On the basis of an understanding of the role of the carboxy group in iridium-catalyzed asymmetric hydrogenation reactions, we developed a carboxy-group-directed strategy for asymmetric hydrogenation of olefins. Using this strategy, we hydrogenated several challenging olefin substrates, such as β,γ-unsaturated carboxylic acids, 1,1-diarylethenes, 1,1-dialkylethenes, and 1-alkyl styrenes in high yield and with excellent enantioselectivity. All these iridium-catalyzed asymmetric hydrogenation reactions feature high turnover numbers (up to 10000) and turnover frequencies (up to 6000 h-1), excellent enantioselectivities (greater than 95% ee with few exceptions), low hydrogen pressure (<12 atm), and operational simplicity. These features make chiral iridium catalysts superior or comparable to well-established chiral ruthenium and rhodium catalysts for asymmetric hydrogenation of unsaturated carboxylic acids. A number of chiral natural products and pharmaceuticals have been prepared by concise routes involving an iridium-catalyzed asymmetric hydrogenation of an unsaturated carboxylic acid as a key step. As part of a mechanistic study of iridium-catalyzed asymmetric hydrogenation of unsaturated acids, we isolated, for the first time, the migratory insertion intermediate in the iridium-catalyzed asymmetric hydrogenation of olefins, and this result strongly supports the involvement of an Ir(III)/Ir(V) catalytic cycle. The rigid, bulky scaffold of the chiral spiro-P,N-ligands of the catalysts not only prevents them from undergoing deactivating aggregation under the hydrogenation conditions but also is responsible for the efficient chiral induction. The carboxy group of the substrate acts as an anchor to ensure coordination of the substrate to the iridium center of the catalyst during the reaction and makes the hydrogenation proceed smoothly.


Liquid-chromatographic determination of zomepirac in serum and plasma.
We describe a determination of zomepirac concentration in plasma and serum by reversed-phase "high-performance" liquid chromatography. The assay requires 1.0 mL of sample and involves diethyl ether extraction of zomepirac from an acidified sample, followed by concentration and injection into a liquid chromatograph. The column effluent is monitored at 330 nm. Retention times of zomepirac and the internal standard (tolmetin) are 3.8 and 2.7 min, respectively. The lower limit of detection for zomepirac in serum or plasma is 0.05 mg/L. Within-day precision (CV) of analysis in plasma or serum with zomepirac added (0.1-10.0 mg/L) ranged from 1.4 to 6.7%; between-day CV varied from 1.4 to 7.5%. Analytical recovery of zomepirac (1.0 mg/L) from serum and plasma was 77.6 (SD 3.5)% and 80.4 (SD 5.2)%, respectively. Numerous commonly coadministered drugs did not interfere. The elimination half-life of the drug was 1.8 h, and the peak plasma concentration ranged from 1.1 to 2.4 mg/L. Peak and trough concentrations measured throughout five days of therapy imply no accumulation.


A longitudinal study of health service utilization for diabetes patients in a care coordination home-telehealth programme.
We assessed the effects of a care coordination home-telehealth (CCHT) programme on health service utilization. The subjects (n = 387 for each group) were US veterans diagnosed with diabetes and followed for 48 months. The service utilization measures were numbers of all-cause inpatient stays and outpatient visits during the follow-up period. We used generalized linear mixed models to estimate the adjusted effects of the CCHT programme on service use over time. Compared with the controls, the CCHT clients were less likely to be admitted for inpatient care during the first (P < 0.001) and second (P < 0.01) six-month follow-up period, and were consistently more likely to visit outpatient clinics (P < 0.001) during the whole 48-month follow-up period. The likelihood of increase in outpatient utilization tended to decline over time. The findings suggest that the CCHT programme helped to reduce overall inpatient and outpatient use by the clients over the 48-month follow-up period.


Consequences of mating with siblings and nonsiblings on the reproductive success in a leaf beetle.
Choosing a suitable mating partner is crucial for the fitness of an individual, whereby mating with siblings often results in inbreeding depression. We studied consequences of mating with siblings versus nonsiblings in the mustard leaf beetle, Phaedon cochleariae (Coleoptera: Chrysomelidae), on lifetime reproductive traits. Furthermore, we analyzed whether cuticular hydrocarbon (CHC) profiles are family specific and could potentially influence the mating behavior of young adults. We hypothesized a reduced reproductive success of females mated with siblings and a more rapid mating of males with nonsiblings. The hatching rate from eggs of sibling pairs was lower compared to that of nonsibling pairs, pointing to inbreeding depression. Furthermore, the number of eggs laid by females decreased over time in both sibling and nonsibling pairs. Interestingly, the CHC profiles and the body mass differed between families. However, the beetles did not avoid siblings and accepted them as readily as nonsiblings for mating in no-choice tests. In summary, although it had negative consequences to mate a sibling and although siblings could potentially be recognized by their CHC profiles, the beetles did not show a delayed mating with siblings. Our results indicate that P. cochleariae beetles have not developed a precopulatory mechanism to avoid inbreeding, at least under the test conditions applied here. We predict that instead a polyandrous mating system and/or postcopulatory mechanisms might have evolved in this species by which inbreeding costs can be reduced.


High-density lipoprotein, but not low-density lipoprotein cholesterol levels influence short-term prognosis after acute coronary syndrome: results from the MIRACL trial.
Patients with acute coronary syndrome (ACS) in the Myocardial Ischaemia Reduction with Aggressive Cholesterol Lowering (MIRACL) study had diminished cardiovascular events after 16 weeks of treatment of atorvastatin 80 mg daily. We determined whether plasma lipoproteins at baseline and then at 6 weeks after randomization predicted clinical outcome. Cox proportional hazards models were constructed to determine relations between lipoproteins and clinical endpoint events. Baseline LDL cholesterol (LDL-C) did not predict outcome. In contrast, baseline HDL-C predicted outcome with a hazard ratio of 0.986 per mg/dL increment in HDL-C, P<0.001, indicating 1.4% reduction in risk for each 1 mg/dL increase in HDL-C. Atorvastatin treatment profoundly lowered LDL-C, but had minimal effect on HDL-C. Neither Week 6 LDL-C nor absolute change of LDL-C from baseline by Week 6 had any significant impact on clinical endpoints occurring between Week 6 and Week 16 after randomization. Plasma HDL-C, but not LDL-C, measured in the initial stage of ACS predicts the risk of recurrent cardiovascular events over the ensuing 16 weeks. LDL-C reduction does not account for the clinical risk reduction with atorvastatin treatment after ACS. This finding may suggest that the clinical benefit of atorvastatin after ACS is mediated by qualitative changes in the LDL particle and/or by non-lipid (pleiotropic) effects of the drug.


Serotonin reduction in post-acute sequelae of viral infection.
Post-acute sequelae of COVID-19 (PASC, "Long COVID") pose a significant global health challenge. The pathophysiology is unknown, and no effective treatments have been found to date. Several hypotheses have been formulated to explain the etiology of PASC, including viral persistence, chronic inflammation, hypercoagulability, and autonomic dysfunction. Here, we propose a mechanism that links all four hypotheses in a single pathway and provides actionable insights for therapeutic interventions. We find that PASC are associated with serotonin reduction. Viral infection and type I interferon-driven inflammation reduce serotonin through three mechanisms: diminished intestinal absorption of the serotonin precursor tryptophan; platelet hyperactivation and thrombocytopenia, which impacts serotonin storage; and enhanced MAO-mediated serotonin turnover. Peripheral serotonin reduction, in turn, impedes the activity of the vagus nerve and thereby impairs hippocampal responses and memory. These findings provide a possible explanation for neurocognitive symptoms associated with viral persistence in Long COVID, which may extend to other post-viral syndromes.


Trajectory and outcomes of speech language therapy in the Prader-Willi syndrome (PWS): case report.
The aim of this study was to describe the trajectory and the outcomes of speech-language therapy in Prader-Willi syndrome through a longitudinal study of the case of an 8 year-old boy, along four years of speech-language therapy follow-up. The therapy sessions were filmed and documental analysis of information from the child's records regarding anamnesis, evaluation and speech-language therapy reports and multidisciplinary evaluations were carried out. The child presented typical characteristics of Prader-Willi syndrome, such as obesity, hyperfagia, anxiety, behavioral problems and self aggression episodes. Speech-language pathology evaluation showed orofacial hypotony, sialorrhea, hypernasal voice, cognitive deficits, oral comprehension difficulties, communication using gestures and unintelligible isolated words. Initially, speech-language therapy had the aim to promote the language development emphasizing social interaction through recreational activities. With the evolution of the case, the main focus became the development of conversation and narrative abilities. It were observed improvements in attention, symbolic play, social contact and behavior. Moreover, there was an increase in vocabulary, and evolution in oral comprehension and the development of narrative abilities. Hence, speech-language pathology intervention in the case described was effective in different linguistic levels, regarding phonological, syntactic, lexical and pragmatic abilities.


Single-cell enzyme concentrations, kinetics, and inhibition analysis using high-density hydrodynamic cell isolation arrays.
High-quality single-cell data are required for a quantitative systems biology description of cellular function. However, data of this type are difficult and time-consuming to collect using traditional techniques. We present a robust and simple microfluidic method for trapping single cells in large arrays to address this problem. Ordered single-cell isolation arrays allow for high-density microscopic analysis with simplified image processing. Moreover, for fluorescent assays, on-chip sample preparation (e.g., fluorescent labeling, washing) can be performed, as opposed to manual intensive operations of incubation, centrifugation, and resuspension in previous techniques-saving time and reagents. This technology was applied to determine novel single-cell enzyme kinetics for three different cell types (HeLa, 293T, Jurkat). A kinetic model of this process predicted this varied response was due to variation in the concentration of carboxylesterase between cell types. Nordihydroguaiaretic acid (NDGA) was also characterized as an inhibitor of carboxylesterases. For HeLa cells, 20 nM of the 50 nM total carboxylesterases was unaffected by NDGA. This type of analysis could be directly applied to quantify a variety of intracellular enzymes with available fluorogenic substrates.


A widespread silent polymorphism of human carbonic anhydrase III (31 Ile in equilibrium Val): implications for evolutionary genetics.
During amino acid sequence studies of carbonic anhydrase (CA) III, purified from a pool of human skeletal muscles, and electrophoretically undetectable (silent) variation was found at residue 31 which was either valine and/or isoleucine. To distinguish a simple allelic polymorphism from more complex models involving gene duplication, 11 separate CA III samples were purified from individuals of different age and racial backgrounds. Peptide mapping by high performance liquid chromatography and sequencing indicated that four were homozygous for 31-Val, three homozygous for 31-Ile and four were apparent heterozygotes. Since the ratio of Val/Ile at residue 31 was approximately 1.0 in the heterozygotes, the present observations are consistent with a simple allelic polymorphism model. Despite the small sample size, there are preliminary indications that the gene frequencies may differ among racial groups. The finding of this silent allelic polymorphism together with the finding of an electrophoretically detectable polymorphism of CA II permits us to test the linkage of the CA II and CA III genes which appear to have been formed by gene duplication more than 300 million years ago. The possibility that the Val/Ile variation may represent a neutral mutation is discussed.


Identification of New MmpL3 Inhibitors by Untargeted and Targeted Mutant Screens Defines MmpL3 Domains with Differential Resistance.
The Mycobacterium tuberculosis mycolate flippase MmpL3 has been the proposed target for multiple inhibitors with diverse chemical scaffolds. This diversity in chemical scaffolds has made it difficult to predict compounds that inhibit MmpL3 without whole-genome sequencing of isolated resistant mutants. Here, we describe the identification of four new inhibitors that select for resistance mutations in mmpL3. Using these resistant mutants, we conducted a targeted whole-cell phenotypic screen of 163 novel M. tuberculosis growth inhibitors for differential growth inhibition of wild-type M. tuberculosis compared to the growth of a pool of 24 unique mmpL3 mutants. The screen successfully identified six additional putative MmpL3 inhibitors. The compounds were bactericidal both in vitro and against intracellular M. tuberculosisM. tuberculosis cells treated with these compounds were shown to accumulate trehalose monomycolates, have reduced levels of trehalose dimycolate, and displace an MmpL3-specific probe, supporting MmpL3 as the target. The inhibitors were mycobacterium specific, with several also showing activity against the nontuberculous mycobacterial species M. abscessus Cluster analysis of cross-resistance profiles generated by dose-response experiments for each combination of 13 MmpL3 inhibitors against each of the 24 mmpL3 mutants defined two clades of inhibitors and two clades of mmpL3 mutants. Pairwise combination studies of the inhibitors revealed interactions that were specific to the clades identified in the cross-resistance profiling. Additionally, modeling of resistance-conferring substitutions to the MmpL3 crystal structure revealed clade-specific localization of the residues to specific domains of MmpL3, with the clades showing differential resistance. Several compounds exhibited high solubility and stability in microsomes and low cytotoxicity in macrophages, supporting their further development. The combined study of multiple mutants and novel compounds provides new insights into structure-function interactions of MmpL3 and small-molecule inhibitors.


Vaccine innovations for emerging infectious diseases-a symposium report.
Vaccines have been incredibly successful at stemming the morbidity and mortality of infectious diseases worldwide. However, there are still no effective vaccines for many serious and potentially preventable infectious diseases. Advances in vaccine technology, including new delivery methods and adjuvants, as well as progress in systems biology and an increased understanding of the human immune system, hold the potential to address these issues. In addition, maternal immunization has opened an avenue to address infectious diseases in neonates and very young infants. This report summarizes the presentations from a 1-day symposium at the New York Academy of Sciences entitled "Innovative Vaccines against Resistant Infectious Diseases and Emerging Threats," held on May 20, 2019.


Cancer gene prioritization by integrative analysis of mRNA expression and DNA copy number data: a comparative review.
A variety of genome-wide profiling techniques are available to investigate complementary aspects of genome structure and function. Integrative analysis of heterogeneous data sources can reveal higher level interactions that cannot be detected based on individual observations. A standard integration task in cancer studies is to identify altered genomic regions that induce changes in the expression of the associated genes based on joint analysis of genome-wide gene expression and copy number profiling measurements. In this review, we highlight common approaches to genomic data integration and provide a transparent benchmarking procedure to quantitatively compare method performances in cancer gene prioritization. Algorithms, data sets and benchmarking results are available at http://intcomp.r-forge.r-project.org.


The bivalent-cation dependence of phosphatidylinositol synthesis in a cell-free system from lymphocytes.
The bivalent-cation requirements of two enzymes involved in phosphatidylinositol synthesis were defined for pig lymphocyte membranes using a citric acid buffer. CTP:phosphatidic acid cytidylyltransferase (EC 22.214.171.124) is activated by free Mn2+ concentrations above 20nM and by free Mg2+ concentrations above 10 microM. When activated by Mg2+, the enzyme is weakly inhibited by Ca2+ (Ki greater than 250 microM), but Ca2+ has no effect when Mn2+ is used to stimulate CDP-diacylglycerol synthesis. The synthesis of phosphatidylinositol from phosphatidic acid is also stimulated by Mn2+ and Mg2+ concentrations similar to those above and is inhibited by free Ca2+ concentrations above 500nM, probably by its action on CDP-diacylglycerol:inositol 3-phosphatidyltransferase (EC 188.8.131.52). Taken together, these studies suggest that under physiological conditions phosphatidylinositol synthesis is activated by Mg2+ and it is possible that it is further regulated by the free concentrations of Ca2+ and/or Mn2+.


Abrupt induction of a membrane digestive enzyme by its intraintestinal substrate.
The regulation of amino-oligopeptidase (AOP), an intestinal brush border hydrolase essential for the surface digestion of peptide nutrients, was examined in rats in vivo. Short-term (30-minute) intraintestinal perfusion of a tetrapeptide substrate, Gly-Leu-Gly-Gly, or a synthetic substrate, leucyl-beta-naphthylamide, induced a doubling in the incorporation of [3H]leucine into the AOP in association with intracellular membranes. The subsequent conversion of AOP from nascent to mature enzyme and its membrane-associated transport to the brush border occurred at normal rates.