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BACKGROUND: Disparities in survival and clinical outcomes between African American and White patients with breast cancer (BC) are well documented, but African American patients have not been well represented in randomized clinical trials of CDK4/6 inhibitors. Real-world studies can provide evidence for effective treatment strategies for underreported patient populations. PATIENTS AND METHODS: This retrospective analysis of African American patients with HR+/HER2- metastatic breast cancer (mBC) from the Flatiron Health longitudinal database evaluated treatments for patients with BC in routine clinical practice in the US. Patients initiated first-line therapy with palbociclib plus an aromatase inhibitor (AI) or AI alone between February 2015 and March 2020. Outcomes assessed included overall survival (OS) and real-world progression-free survival (rwPFS) until September 2020. RESULTS: Of 270 eligible patients, 127 (median age 64 years) were treated with palbociclib + AI, and 143 (median age 68 years) were treated with an AI. Median follow-up was 24.0 months for palbociclib + AI and 18.2 months for AI-treated patients. Median OS was not reached (NR; 95% CI, 38.2-NR) in the palbociclib + AI group versus 28.2 months (95% CI, 19.2-52.8) in the AI group (adjusted HR, 0.56; 95% CI, 0.36-0.89; P = .013). Median rwPFS was 18.0 months (95% CI, 12.4-26.7) in the palbociclib + AI group and 10.5 months (95% CI, 7.0-13.4) in the AI group (adjusted HR, 0.74; 95% CI, 0.47-1.17; P = .199). CONCLUSION: This comparative analysis of palbociclib + AI versus AI alone indicates that palbociclib combined with endocrine therapy in the first line is associated with improved effectiveness for African American patients with HR+/HER2- mBC in real-world settings. TRIAL NUMBER: NCT05361655.

Purpose: To explore the journey taken by patients in a range of different countries to manage their dry eye symptoms. Method: Members of the general public who responded positively to the question “Do your eyes ever feel dry?” completed a questionnaire describing their demographics, the impact of their symptomology, the advice they have received and the management options they have tried. The Ocular Surface Disease Index (OSDI) questionnaire was also completed. Results: A total of 916 individuals (Canada = 235, Mexico = 127, New Zealand = 157, Taiwan = 246, UK = 151) of similar age distribution (median 38 years, IQR: 27–50) completed the survey. The reported duration of symptoms was longest in Canada (median 4 years, range 2–10) and least in Taiwan (2 years, range 1–3; p < 0.001), and similar trends were observed for symptom severity (p = 0.001). However, there was no statistically significant difference between countries with respect to the impact of symptoms on quality of life (median 3/10; p = 0.08). Less than half of the individuals in any country had consulted with a health professional. About half had tried a treatment for their dry eye symptoms, with artificial tears being the most common treatment, followed by warm compresses, and both therapies were rated as reasonably effective (median 5−7/10). Conclusion: Many people with dry eye symptoms are not consulting health care professionals who can confirm the diagnosis, exclude differential diagnoses, and offer a wide range of treatments targeted at the dry eye subtype.

Purpose: To explore the journey taken by patients in a range of different countries to manage their dry eye symptoms. Method: Members of the general public who responded positively to the question “Do your eyes ever feel dry?” completed a questionnaire describing their demographics, the impact of their symptomology, the advice they have received and the management options they have tried. The Ocular Surface Disease Index (OSDI) questionnaire was also completed. Results: A total of 916 individuals (Canada = 235, Mexico = 127, New Zealand = 157, Taiwan = 246, UK = 151) of similar age distribution (median 38 years, IQR: 27–50) completed the survey. The reported duration of symptoms was longest in Canada (median 4 years, range 2–10) and least in Taiwan (2 years, range 1–3; p < 0.001), and similar trends were observed for symptom severity (p = 0.001). However, there was no statistically significant difference between countries with respect to the impact of symptoms on quality of life (median 3/10; p = 0.08). Less than half of the individuals in any country had consulted with a health professional. About half had tried a treatment for their dry eye symptoms, with artificial tears being the most common treatment, followed by warm compresses, and both therapies were rated as reasonably effective (median 5−7/10). Conclusion: Many people with dry eye symptoms are not consulting health care professionals who can confirm the diagnosis, exclude differential diagnoses, and offer a wide range of treatments targeted at the dry eye subtype.

BackgroundDepression and anxiety are leading causes of disability worldwide. Though effective treatments exist, depression and anxiety remain undertreated. Blended care psychotherapy, combining the scalability of online interventions with the personalization and engagement of a live therapist, is a promising approach for increasing access to evidence-based care.ObjectivesTo evaluate the effectiveness and individual contribution of two components - i) digital tools and ii) video-based therapist-led sessions - in a blended care CBT-based intervention under real world conditions.MethodsA retrospective cohort design was used to analyze N = 1372 US-based individuals who enrolled in blended care psychotherapy. Of these, at baseline, 761 participants had depression symptoms in the clinical range (based on PHQ-9), and 1254 had anxiety symptoms in the clinical range (based on GAD-7). Participants had access to the program as a mental health benefit offered by their employer. The CBT-based blended care psychotherapy program consisted of regular video sessions with therapists, complemented by digital lessons and digital exercises assigned by the clinician and completed in between sessions. Depression and anxiety levels and clients' treatment engagement were tracked throughout treatment. A 3-level individual growth curve model incorporating time-varying covariates was utilized to examine symptom trajectories of PHQ-9 scores (for those with clinical range of depression at baseline) and GAD-7 scores (for those with clinical range of anxiety at baseline).ResultsOn average, individuals exhibited a significant decline in depression and anxiety symptoms during the initial weeks of treatment (P < .001), and a continued decline over subsequent weeks at a slower rate (P < .001). Engaging in a therapy session in a week was associated with lower GAD-7 (b = -0.81) and PHQ-9 (b = -1.01) scores in the same week, as well as lower GAD-7 (

BackgroundBoth the gut microbiota and chronic viral infections have profound effects on host immunity, but interactions between these influences have been only superficially explored. Cytomegalovirus (CMV), for example, infects approximately 80% of people globally and drives significant changes in immune cells. Similarly, certain gut-resident bacteria affect T-cell development in mice and nonhuman primates. It is unknown if changes imposed by CMV on the intestinal microbiome contribute to immunologic effects of the infection.ResultsWe show that rhesus cytomegalovirus (RhCMV) infection is associated with specific differences in gut microbiota composition, including decreased abundance of Firmicutes, and that the extent of microbial change was associated with immunologic changes including the proliferation, differentiation, and cytokine production of CD8+ T cells. Furthermore, RhCMV infection disrupted the relationship between short-chain fatty acid producers and Treg/Th17 balance observed in seronegative animals, showing that some immunologic effects of CMV are due to disruption of previously existing host-microbe relationships.ConclusionsGut microbes have an important influence on health and disease. Diet is known to shape the microbiota, but the influence of concomitant chronic viral infections is unclear. We found that CMV influences gut microbiota composition to an extent that is correlated with immunologic changes in the host. Additionally, pre-existing correlations between immunophenotypes and gut microbes can be subverted by CMV infection. Immunologic effects of CMV infection on the host may therefore be mediated by two different mechanisms involving gut microbiota. Video Abstract.

Data on real-world effectiveness of cyclin-dependent kinase 4/6 inhibitor combination therapy versus endocrine therapy alone are limited. The Flatiron Health Analytic Database was used to assess overall survival (OS) in patients with hormone receptor-positive/human epidermal growth factor receptor 2-negative (HR+/HER2-) metastatic breast cancer (MBC) treated with first-line palbociclib plus an aromatase inhibitor (AI) versus an AI alone in routine US clinical practice. In total, 2888 patients initiated treatment during February 3, 2015-March 31, 2020, with a potential ≥6-month follow-up (cutoff date, September 30, 2020). After stabilized inverse probability treatment weighting, median OS (95% CI) is significantly longer among palbociclib versus AI recipients (49.1 [45.2-57.7] versus 43.2 [37.6-48.0] months; hazard ratio, 0.76 [95% CI, 0.65-0.87]; P < 0.0001). Progression-free survival (95% CI) is 19.3 (17.5-20.7) versus 13.9 (12.5-15.2) months, respectively (hazard ratio, 0.70 [95% CI, 0.62-0.78]; P < 0.0001). These data support first-line palbociclib plus an AI treatment for HR+/HER2- MBC.(Trial number NCT05361655).

Purpose: To explore the journey taken by patients in a range of different countries to manage their dry eye symptoms. Method: Members of the general public who responded positively to the question “Do your eyes ever feel dry?” completed a questionnaire describing their demographics, the impact of their symptomology, the advice they have received and the management options they have tried. The Ocular Surface Disease Index (OSDI) questionnaire was also completed. Results: A total of 916 individuals (Canada = 235, Mexico = 127, New Zealand = 157, Taiwan = 246, UK = 151) of similar age distribution (median 38 years, IQR: 27–50) completed the survey. The reported duration of symptoms was longest in Canada (median 4 years, range 2–10) and least in Taiwan (2 years, range 1–3; p < 0.001), and similar trends were observed for symptom severity (p = 0.001). However, there was no statistically significant difference between countries with respect to the impact of symptoms on quality of life (median 3/10; p = 0.08). Less than half of the individuals in any country had consulted with a health professional. About half had tried a treatment for their dry eye symptoms, with artificial tears being the most common treatment, followed by warm compresses, and both therapies were rated as reasonably effective (median 5−7/10). Conclusion: Many people with dry eye symptoms are not consulting health care professionals who can confirm the diagnosis, exclude differential diagnoses, and offer a wide range of treatments targeted at the dry eye subtype.

Purpose: To explore the journey taken by patients in a range of different countries to manage their dry eye symptoms. Method: Members of the general public who responded positively to the question “Do your eyes ever feel dry?” completed a questionnaire describing their demographics, the impact of their symptomology, the advice they have received and the management options they have tried. The Ocular Surface Disease Index (OSDI) questionnaire was also completed. Results: A total of 916 individuals (Canada = 235, Mexico = 127, New Zealand = 157, Taiwan = 246, UK = 151) of similar age distribution (median 38 years, IQR: 27–50) completed the survey. The reported duration of symptoms was longest in Canada (median 4 years, range 2–10) and least in Taiwan (2 years, range 1–3; p < 0.001), and similar trends were observed for symptom severity (p = 0.001). However, there was no statistically significant difference between countries with respect to the impact of symptoms on quality of life (median 3/10; p = 0.08). Less than half of the individuals in any country had consulted with a health professional. About half had tried a treatment for their dry eye symptoms, with artificial tears being the most common treatment, followed by warm compresses, and both therapies were rated as reasonably effective (median 5−7/10). Conclusion: Many people with dry eye symptoms are not consulting health care professionals who can confirm the diagnosis, exclude differential diagnoses, and offer a wide range of treatments targeted at the dry eye subtype.

Connie Chen,1 Mary Ayers,1 Judy H Squires,2 James E Squires1 1Division of Gastroenterology, Hepatology and Nutrition, UPMC Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA; 2Department of Radiology, UPMC Children’s Hospital of Pittsburgh, Pittsburgh, PA, USACorrespondence: James E Squires, Division of Gastroenterology, Hepatology and Nutrition, Children’s Hospital of Pittsburgh, Pittsburgh, PA, 15224, Tel +1 412-692-5180, Fax +1 412-692-7355, Email James.Squires2@chp.eduAbstract: Sarcopenia, a pathologic deficiency of muscle mass and function, has emerged as an important secondary feature of many chronic disease states. For adults with end stage liver disease, there are multiple mechanisms which contribute to sarcopenia and its presence has proven to be an important predictor of morbidity and mortality. In children, there are only a limited number of reports which investigate the role of sarcopenia in liver disease. These studies, which are discussed and summarized in this review, report small, single-center analyses with dissimilar study cohorts and varying clinical definitions. Still, children meeting the study entry criteria have sarcopenia with a reported prevalence of 24– 70%. When assessed, sarcopenia appears to be associated with more severe disease but is independent of the Pediatric End-Stage Liver Disease (PELD) score and does not correlate with age, gender, or traditional anthropometric measures such as weight, height, weight-for-height, or body mass index (BMI). While individual studies may identify sarcopenia as a statistically significant risk factor for certain post-transplant outcomes such as longer ICU stay, longer duration of intubation, repeat operation, development of serious infection, longer hospital stay, death, or long-term growth failure, such associations are not consistently replicated across studies. Finally, although various methods of muscle mass quantification are utilized, the most reported is the total psoas muscle surf

Purpose: A survey in 2015 identified a high level of eye care practitioner concern about myopia with a reported moderately high level of activity, but the vast majority still prescribed single vision interventions to young myopes. This research aimed to update these findings 4 years later. Methods: A self-administrated, internet-based questionnaire was distributed in eight languages, through professional bodies to eye care practitioners globally. The questions examined: awareness of increasing myopia prevalence, perceived efficacy of available strategies and adoption levels of such strategies, and reasons for not adopting specific strategies. Results: Of the 1336 respondents, concern was highest (9.0 ± 1.6; p < 0.001) in Asia and lowest (7.6 ± 2.2; p < 0.001) in Australasia. Practitioners from Asia also considered their clinical practice of myopia control to be the most active (7.7 ± 2.3; p < 0.001), the North American practitioners being the least active (6.3 ± 2.9; p < 0.001). Orthokeratology was perceived to be the most effective method of myopia control, followed by pharmaceutical approaches and approved myopia control soft contact lenses (p < 0.001). Although significant intra-regional differences existed, overall, most practitioners did not consider single-vision distance under-correction to be an effective strategy for attenuating myopia progression (79.6 %), but prescribed single vision spectacles or contact lenses as the primary mode of correction for myopic patients (63.6 ± 21.8 %). The main justifications for their reluctance to prescribe alternatives to single vision refractive corrections were increased cost (20.6 %) and inadequate information (17.6 %). Conclusions: While practitioner concern about myopia and the reported level of activity have increased over the last 4 years, the vast majority of eye care clinicians still prescribe single vision interventions to young myopes. With recent global consensus evidence-based guidelines having been published

Purpose: A survey in 2015 identified a high level of eye care practitioner concern about myopia with a reported moderately high level of activity, but the vast majority still prescribed single vision interventions to young myopes. This research aimed to update these findings 4 years later. Methods: A self-administrated, internet-based questionnaire was distributed in eight languages, through professional bodies to eye care practitioners globally. The questions examined: awareness of increasing myopia prevalence, perceived efficacy of available strategies and adoption levels of such strategies, and reasons for not adopting specific strategies. Results: Of the 1336 respondents, concern was highest (9.0 ± 1.6; p < 0.001) in Asia and lowest (7.6 ± 2.2; p < 0.001) in Australasia. Practitioners from Asia also considered their clinical practice of myopia control to be the most active (7.7 ± 2.3; p < 0.001), the North American practitioners being the least active (6.3 ± 2.9; p < 0.001). Orthokeratology was perceived to be the most effective method of myopia control, followed by pharmaceutical approaches and approved myopia control soft contact lenses (p < 0.001). Although significant intra-regional differences existed, overall, most practitioners did not consider single-vision distance under-correction to be an effective strategy for attenuating myopia progression (79.6 %), but prescribed single vision spectacles or contact lenses as the primary mode of correction for myopic patients (63.6 ± 21.8 %). The main justifications for their reluctance to prescribe alternatives to single vision refractive corrections were increased cost (20.6 %) and inadequate information (17.6 %). Conclusions: While practitioner concern about myopia and the reported level of activity have increased over the last 4 years, the vast majority of eye care clinicians still prescribe single vision interventions to young myopes. With recent global consensus evidence-based guidelines having been published

Peter C Taylor, 1 Neil Betteridge, 2 T Michelle Brown, 3 John Woolcott, 4 Alan J Kivitz, 5 Cristiano Zerbini, 6 Diane Whalley, 7 Oyebimpe Olayinka-Amao, 3 Connie Chen, 8 Palle Dahl, 9 Dario Ponce de Leon, 10 David Gruben, 11 Lara Fallon 12 1Botnar Research Centre, University of Oxford, Oxford, UK; 2Neil Betteridge Associates, London, UK; 3Patient-Centered Outcomes Assessment Group, RTI Health Solutions, Research Triangle Park, NC, USA; 4Patient and Health Impact, Health Economics and Outcomes Research, Pfizer Inc, Collegeville, PA, USA; 5Altoona Center for Clinical Research, Duncansville, PA, USA; 6Department of Rheumatology, Centro Paulista De Investigação Clinica, São Paulo, Brazil; 7Patient-Centered Outcomes Assessment Group, RTI Health Solutions, Manchester, UK; 8Xeljanz, Rheumatology, Inflammation & Immunology Medical Affairs, Pfizer Inc, New York, NY, USA; 9Medical Affairs, International Developed Markets, Inflammation & Immunology, Pfizer Inc, Ballerup, Denmark; 10Medical Affairs Latin-America, Pfizer Inc, New York, NY, USA; 11Statistical Research and Data Science Center, Pfizer Inc, Groton, CT, USA; 12Global Medical Affairs, Pfizer Inc, Montreal, QC, CanadaCorrespondence: Peter C TaylorBotnar Research Centre, University of Oxford, Windmill Road, Headington, Oxford OX3 7LD, UKTel +441865 227323Email peter.taylor@kennedy.ox.ac.ukPurpose: Current knowledge of the reasons for patients’ preference for rheumatoid arthritis (RA) treatment modes is limited. This study was designed to identify preferences for four treatment modes, and to obtain in-depth information on the reasons for these preferences.Patients and Methods: In this multi-national, cross-sectional, qualitative study, in-depth interviews were conducted with adult patients with RA in the United States, France, Germany, Italy, Spain, Switzerland, the United Kingdom, and Brazil. Patients’ strength of preference was evaluated using a 100-p

Purpose: A survey in 2015 identified a high level of eye care practitioner concern about myopia with a reported moderately high level of activity, but the vast majority still prescribed single vision interventions to young myopes. This research aimed to update these findings 4 years later. Methods: A self-administrated, internet-based questionnaire was distributed in eight languages, through professional bodies to eye care practitioners globally. The questions examined: awareness of increasing myopia prevalence, perceived efficacy of available strategies and adoption levels of such strategies, and reasons for not adopting specific strategies. Results: Of the 1336 respondents, concern was highest (9.0 ± 1.6; p < 0.001) in Asia and lowest (7.6 ± 2.2; p < 0.001) in Australasia. Practitioners from Asia also considered their clinical practice of myopia control to be the most active (7.7 ± 2.3; p < 0.001), the North American practitioners being the least active (6.3 ± 2.9; p < 0.001). Orthokeratology was perceived to be the most effective method of myopia control, followed by pharmaceutical approaches and approved myopia control soft contact lenses (p < 0.001). Although significant intra-regional differences existed, overall, most practitioners did not consider single-vision distance under-correction to be an effective strategy for attenuating myopia progression (79.6 %), but prescribed single vision spectacles or contact lenses as the primary mode of correction for myopic patients (63.6 ± 21.8 %). The main justifications for their reluctance to prescribe alternatives to single vision refractive corrections were increased cost (20.6 %) and inadequate information (17.6 %). Conclusions: While practitioner concern about myopia and the reported level of activity have increased over the last 4 years, the vast majority of eye care clinicians still prescribe single vision interventions to young myopes. With recent global consensus evidence-based guidelines having been published

Purpose: A survey in 2015 identified a high level of eye care practitioner concern about myopia with a reported moderately high level of activity, but the vast majority still prescribed single vision interventions to young myopes. This research aimed to update these findings 4 years later. Methods: A self-administrated, internet-based questionnaire was distributed in eight languages, through professional bodies to eye care practitioners globally. The questions examined: awareness of increasing myopia prevalence, perceived efficacy of available strategies and adoption levels of such strategies, and reasons for not adopting specific strategies. Results: Of the 1336 respondents, concern was highest (9.0 ± 1.6; p < 0.001) in Asia and lowest (7.6 ± 2.2; p < 0.001) in Australasia. Practitioners from Asia also considered their clinical practice of myopia control to be the most active (7.7 ± 2.3; p < 0.001), the North American practitioners being the least active (6.3 ± 2.9; p < 0.001). Orthokeratology was perceived to be the most effective method of myopia control, followed by pharmaceutical approaches and approved myopia control soft contact lenses (p < 0.001). Although significant intra-regional differences existed, overall, most practitioners did not consider single-vision distance under-correction to be an effective strategy for attenuating myopia progression (79.6 %), but prescribed single vision spectacles or contact lenses as the primary mode of correction for myopic patients (63.6 ± 21.8 %). The main justifications for their reluctance to prescribe alternatives to single vision refractive corrections were increased cost (20.6 %) and inadequate information (17.6 %). Conclusions: While practitioner concern about myopia and the reported level of activity have increased over the last 4 years, the vast majority of eye care clinicians still prescribe single vision interventions to young myopes. With recent global consensus evidence-based guidelines having been published

The HIV reservoir forming at the earliest stages of infection is likely composed of CCR5+ cells, because these cells are the targets of transmissible virus. Restriction of the CCR5+ reservoir, particularly in the gut, may be needed for subsequent cure attempts. Strategies for killing or depleting CCR5+ cells have been described, but none have been tested in vivo in nonhuman primates, and the extent of achievable depletion from tissues is not known. In this study we investigate the efficacy of two novel cytotoxic treatments for targeting and eliminating CCR5+ cells in young rhesus macaques. The first, an immunotoxin consisting of the endogenous CCR5 ligand RANTES fused with Pseudomonas exotoxin (RANTES-PE38), killed CCR5+ lamina propria lymphocytes (LPLs) ex vivo, but had no detectable effect on CCR5+ LPLs in vivo. The second, a primatized bispecific antibody for CCR5 and CD3, depleted all CCR5+ cells from blood and the vast majority of such cells from the colonic mucosa (up to 96% of CD4+CCR5+). Absence of CCR5-expressing cells from blood endured for at least 1 week, while CCR5+ cells in colon were substantially replenished over the same time span. These data open an avenue to investigation of combined early ART treatment and CCR5+ reservoir depletion for cure of HIV-infected infants.

SOX2 is a pan-neural transcription factor expressed in neural precursor cells (NPCs) independently of their regional identity. It plays a crucial role in both neural development and in adult neurogenesis; however the molecular mechanisms underlying its function are poorly understood. Our previous data suggested that SOX2 controls LIN28, an inhibitor of let-7 miRNA biogenesis. We hypothesized that some of the pan-neural functions of SOX2 could be mediated by repression of let-7 miRNA activity. To identify miRNAs with pan-neural SOX2 dependency, I used NPCs with two different regional identities, dorsal and ventral. I modified a previously established human embryonic stem cell (hESC)-derivation protocol (which generated dorsal NPCs) by patterning NPCs with the Sonic hedgehog agonist purmorphamine, yieding ventral NPCs. Analysis of SOX2 targets in dorsal and ventral NPCs in addition to bioinformatics suggested that SOX2 represses the levels of let-7b and let-7i miRNA. Overexpression studies of let-7b and let-7i revealed that let-7b selectively suppresses NPC proliferation with no effect on neuronal differentiation, while let-7i abolishes neuronal differentiation without inhibiting NPC proliferation. These data suggest that the combined effect of let-7b and let-7i overexpression in NPCs photocopies the loss of SOX2 in these cells. Taken together, our results suggest that in our in vitro cultures, SOX2 suppresses the activity of let-7 miRNAs in NPCs. We propose that the function of let- 7 miRNA family downstream of SOX2 may be a general mechanism for controlling both NPC proliferation and neurogenesis in developmental and adult contexts

SOX2 is a pan-neural transcription factor expressed in neural precursor cells (NPCs) independently of their regional identity. It plays a crucial role in both neural development and in adult neurogenesis; however the molecular mechanisms underlying its function are poorly understood. Our previous data suggested that SOX2 controls LIN28, an inhibitor of let-7 miRNA biogenesis. We hypothesized that some of the pan-neural functions of SOX2 could be mediated by repression of let-7 miRNA activity. To identify miRNAs with pan-neural SOX2 dependency, I used NPCs with two different regional identities, dorsal and ventral. I modified a previously established human embryonic stem cell (hESC)-derivation protocol (which generated dorsal NPCs) by patterning NPCs with the Sonic hedgehog agonist purmorphamine, yieding ventral NPCs. Analysis of SOX2 targets in dorsal and ventral NPCs in addition to bioinformatics suggested that SOX2 represses the levels of let-7b and let-7i miRNA. Overexpression studies of let-7b and let-7i revealed that let-7b selectively suppresses NPC proliferation with no effect on neuronal differentiation, while let-7i abolishes neuronal differentiation without inhibiting NPC proliferation. These data suggest that the combined effect of let-7b and let-7i overexpression in NPCs photocopies the loss of SOX2 in these cells. Taken together, our results suggest that in our in vitro cultures, SOX2 suppresses the activity of let-7 miRNAs in NPCs. We propose that the function of let- 7 miRNA family downstream of SOX2 may be a general mechanism for controlling both NPC proliferation and neurogenesis in developmental and adult contexts

To gain insight into the cellular and molecular cues that promote neurovascular co-patterning at the earliest stages of human embryogenesis, we developed a human embryonic stem cell model to mimic the developing epiblast. Contact of ectoderm-derived neural cells with mesoderm-derived vasculature is initiated via the neural crest (NC), not the neural tube (NT). Neurovascular co-patterning then ensues with specification of NC toward an autonomic fate requiring vascular endothelial cell (EC)-secreted nitric oxide (NO) and direct contact with vascular smooth muscle cells (VSMCs) via T-cadherin-mediated homotypic interactions. Once a neurovascular template has been established, NT-derived central neurons then align themselves with the vasculature. Our findings reveal that, in early human development, the autonomic nervous system forms in response to distinct molecular cues from VSMCs and ECs, providing a model for how other developing lineages might coordinate their co-patterning.

To gain insight into the cellular and molecular cues that promote neurovascular co-patterning at the earliest stages of human embryogenesis, we developed a human embryonic stem cell model to mimic the developing epiblast. Contact of ectoderm-derived neural cells with mesoderm-derived vasculature is initiated via the neural crest (NC), not the neural tube (NT). Neurovascular co-patterning then ensues with specification of NC toward an autonomic fate requiring vascular endothelial cell (EC)-secreted nitric oxide (NO) and direct contact with vascular smooth muscle cells (VSMCs) via T-cadherin-mediated homotypic interactions. Once a neurovascular template has been established, NT-derived central neurons then align themselves with the vasculature. Our findings reveal that, in early human development, the autonomic nervous system forms in response to distinct molecular cues from VSMCs and ECs, providing a model for how other developing lineages might coordinate their co-patterning.

Shukui Qin,1 Feng Bi,2 Jie Jin,3 Ying Cheng,4 Jun Guo,5 Xiubao Ren,6 Yiran Huang,7 Jamal Tarazi,8 Jie Tang,9 Connie Chen,9 Sinil Kim,8 Dingwei Ye10 1Department of Medical Oncology, PLA Cancer Center, Nanjing Bayi Hospital, Nanjing, Jiangsu, People’s Republic of China; 2Department of Medical Oncology, West China Hospital of Sichuan University, Chengdu, Sichuan Province, People’s Republic of China; 3Department of Urology, Peking University First Hospital, Beijing, People’s Republic of China; 4Department of Oncology, Jilin Provincial Cancer Hospital, Changchun, Jilin Province, People’s Republic of China; 5Department of Renal Cancer and Melanoma, Peking University Cancer Hospital/Institute, Beijing, People’s Republic of China; 6Department of Biology Treatment, Tianjin Oncology Hospital, Tianjin, People’s Republic of China; 7Department of Urology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 8Clinical Development, Pfizer Oncology, San Diego, CA, USA; 9Global Outcomes Research, Pfizer Inc., New York, NY, USA; 10Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, People’s Republic of China Background: This registrational trial evaluated the efficacy, safety, and patient-reported outcomes of axitinib versus sorafenib as a second-line treatment in Asian patients with clear-cell metastatic renal cell carcinoma (mRCC). Methods: In this open-label, multicenter study, previously treated Asian patients with clear-cell mRCC were stratified by Eastern Cooperative Oncology Group performance status and prior therapy and randomized in a 2:1 ratio to receive axitinib (5 mg twice daily) or sorafenib (400 mg twice daily). The primary end point was progression-free survival (PFS) assessed by a masked independent review committee.Results: A total of 204 Asian patients received axitinib (n=135) or sorafenib (n=69). Median PFS (95% c

Background Adenosine triphosphate (ATP) is used as an intracellular energy source by all living organisms. It plays a central role in the respiration and metabolism, and is the most important energy supplier in many enzymatic reactions. Its critical role as the energy storage molecule makes it extremely valuable to all cells. Results We report here the detection of extracellular ATP in the cultures of a variety of bacterial species. The levels of the extracellular ATP in bacterial cultures peaked around the end of the log phase and decreased in the stationary phase of growth. Extracellular ATP levels were dependent on the cellular respiration as bacterial mutants lacking cytochrome bo oxidase displayed lower extracellular ATP levels. We have also shown that Escherichia coli (E. coli) and Salmonella actively depleted extracellular ATP and an ATP supplement in culture media enhanced the stationary survival of E. coli and Salmonella. In addition to E. coli and Salmonella the presence of the extracellular ATP was observed in a variety of bacterial species that contain human pathogens such as Acinetobacter, Pseudomonas, Klebsiella and Staphylococcus. Conclusion Our results indicate that extracellular ATP is produced by many bacterial species during growth and extracellular ATP may serve a role in the bacterial physiology.

Background Adenosine triphosphate (ATP) is used as an intracellular energy source by all living organisms. It plays a central role in the respiration and metabolism, and is the most important energy supplier in many enzymatic reactions. Its critical role as the energy storage molecule makes it extremely valuable to all cells. Results We report here the detection of extracellular ATP in the cultures of a variety of bacterial species. The levels of the extracellular ATP in bacterial cultures peaked around the end of the log phase and decreased in the stationary phase of growth. Extracellular ATP levels were dependent on the cellular respiration as bacterial mutants lacking cytochrome bo oxidase displayed lower extracellular ATP levels. We have also shown that Escherichia coli (E. coli) and Salmonella actively depleted extracellular ATP and an ATP supplement in culture media enhanced the stationary survival of E. coli and Salmonella. In addition to E. coli and Salmonella the presence of the extracellular ATP was observed in a variety of bacterial species that contain human pathogens such as Acinetobacter, Pseudomonas, Klebsiella and Staphylococcus. Conclusion Our results indicate that extracellular ATP is produced by many bacterial species during growth and extracellular ATP may serve a role in the bacterial physiology.

Henry J Henk,1 Connie Chen,2 Agnes Benedict,3 Jane Sullivan,1 April Teitelbaum1 1Optum, Eden Prairie, MN, USA; 2Pfizer Inc, New York, NY, USA; 3United BioSource Corporation, London, UK Introduction: Survival and best supportive care (BSC) costs for patients with metastatic renal cell carcinoma (mRCC), after stopping therapy, are poorly characterized yet an important aspect of patient care. This study examined survival and costs associated with BSC after one or two lines of therapy (LOTs) for mRCC. Methods: A retrospective cohort analysis used claims data from commercially insured or Medicare Advantage Prescription Drug (MAPD) plan enrollees of a large United States health plan with an index RCC diagnosis (ICD-9-CM 189.0) between January 1, 2007 and June 30, 2010; initiating any of the following therapies 30 days pre-index date through disenrollment from plan: sunitinib, temsirolimus, sorafenib, bevacizumab, everolimus, pazopanib, cytokines. LOT was identified using prescription fill and administration dates. Health care costs represent health plan- plus patient-paid amounts. Results: The cohort (n = 274) was 73% male, with a mean age of 63.3 years (SD 11.1), with 80% commercially insured (20% MAPD), and 68% starting BSC following one LOT. Mean BSC duration was longer following one than two LOTs (223 [SD 260], 176 [SD 163] days). Median survival from the start of BSC was similar following one and two LOTs (126 and 118 days). Total BSC costs following one and two LOTs averaged US$50,188 (SD $96,984) and $37,295 (SD $51,102). Monthly costs for BSC following one and two LOTs ($10,151 and $10,566) were not substantially lower than costs while on treatment ($14,621 and $16,957). Inpatient hospital costs represented 47% and 49% following one and two LOTs, with ambulatory costs of approximately 36% following each LOT. Conclusion: Our study found similar survival and monthly costs for BSC following either one or two LOTs, with almost half of the cost reflecting inpatient car

Beth Sherrill,1 James A Kaye,2 Rickard Sandin,3 Joseph C Cappelleri,4 Connie Chen51RTI Health Solutions, Biometrics, Research Triangle Park, NC, USA; 2RTI Health Solutions, Epidemiology, Research Triangle Park, NC, USA; 3Pfizer, Global Outcomes Research Sollentuna, Sweden; 4Pfizer, Biostatistics, Groton, CT, USA; 5Pfizer, Global Outcomes Research New York, NY, USAAbstract: Overall survival (OS) is the gold standard in measuring the treatment effect of new drug therapies for cancer. However, practical factors may preclude the collection of unconfounded OS data, and surrogate endpoints are often used instead. Meta-analyses have been widely used for the validation of surrogate endpoints, specifically in oncology. This research reviewed published meta-analyses on the types of surrogate measures used in oncology studies and examined the extent of correlation between surrogate endpoints and OS for different cancer types. A search was conducted in October 2010 to compile available published evidence in the English language for the validation of disease progression-related endpoints as surrogates of OS, based on meta-analyses. We summarize published meta-analyses that quantified the correlation between progression-based endpoints and OS for multiple advanced solid-tumor types. We also discuss issues that affect the interpretation of these findings. Progression-free survival is the most commonly used surrogate measure in studies of advanced solid tumors, and correlation with OS is reported for a limited number of cancer types. Given the increased use of crossover in trials and the availability of second-/third-line treatment options available to patients after progression, it will become increasingly more difficult to establish correlation between effects on progression-free survival and OS in additional tumor types.Keywords: progression endpoints, correlation, cancer

Sufu (Suppressor of Fused), a two-domain protein, plays a critical role in regulating Hedgehog signaling and is conserved from flies to humans. A few bacterial Sufu-like proteins have previously been identified based on sequence similarity to the N-terminal domain of eukaryotic Sufu proteins, but none have been structurally or biochemically characterized and their function in bacteria is unknown. We have determined the crystal structure of a more distantly related Sufu-like homolog, NGO1391 from Neisseria gonorrhoeae, at 1.4 Å resolution, which provides the first biophysical characterization of a bacterial Sufu-like protein. The structure revealed a striking similarity to the N-terminal domain of human Sufu (r.m.s.d. of 2.6 Å over 93% of the NGO1391 protein), despite an extremely low sequence identity of ∼15%. Subsequent sequence analysis revealed that NGO1391 defines a new subset of smaller, Sufu-like proteins that are present in ∼200 bacterial species and has resulted in expansion of the SUFU (PF05076) family in Pfam.

The crystal structures of BB2672 and SPO0826 were determined to resolutions of 1.7 and 2.1 Å by single-wavelength anomalous dispersion and multiple-wavelength anomalous dispersion, respectively, using the semi-automated high-throughput pipeline of the Joint Center for Structural Genomics (JCSG) as part of the NIGMS Protein Structure Initiative (PSI). These proteins are the first structural representatives of the PF06684 (DUF1185) Pfam family. Structural analysis revealed that both structures adopt a variant of the Bacillus chorismate mutase fold (BCM). The biological unit of both proteins is a hexamer and analysis of homologs indicates that the oligomer interface residues are highly conserved. The conformation of the critical regions for oligomerization appears to be dependent on pH or salt concentration, suggesting that this protein might be subject to environmental regulation. Structural similarities to BCM and genome-context analysis suggest a function in amino-acid synthesis.

Membrane-attack complex/perforin (MACPF) proteins are transmembrane pore-forming proteins that are important in both human immunity and the virulence of pathogens. Bacterial MACPFs are found in diverse bacterial species, including most human gut-associated Bacteroides species. The crystal structure of a bacterial MACPF-domain-containing protein BT_3439 (Bth-MACPF) from B. thetaiotaomicron, a predominant member of the mammalian intestinal microbiota, has been determined. Bth-MACPF contains a membrane-attack complex/perforin (MACPF) domain and two novel C-terminal domains that resemble ribonuclease H and interleukin 8, respectively. The entire protein adopts a flat crescent shape, characteristic of other MACPF proteins, that may be important for oligomerization. This Bth-MACPF structure provides new features and insights not observed in two previous MACPF structures. Genomic context analysis infers that Bth-MACPF may be involved in a novel protein-transport or nutrient-uptake system, suggesting an important role for these MACPF proteins, which were likely to have been inherited from eukaryotes via horizontal gene transfer, in the adaptation of commensal bacteria to the host environment.

Proteins with the DUF2063 domain constitute a new Pfam family, PF09836. The crystal structure of a member of this family, NGO1945 from Neisseria gonorrhoeae, has been determined and reveals that the N-terminal DUF2063 domain is likely to be a DNA-binding domain. In conjunction with the rest of the protein, NGO1945 is likely to be involved in transcriptional regulation, which is consistent with genomic neighborhood analysis. Of the 216 currently known proteins that contain a DUF2063 domain, the most significant sequence homologs of NGO1945 (∼40-99% sequence identity) are from various Neisseria and Haemophilus species. As these are important human pathogens, NGO1945 represents an interesting candidate for further exploration via biochemical studies and possible therapeutic intervention.

BT1062 from Bacteroides thetaiotaomicron is a homolog of Mfa2 (PGN0288 or PG0179), which is a component of the minor fimbriae in Porphyromonas gingivalis. The crystal structure of BT1062 revealed a conserved fold that is widely adopted by fimbrial components.

The crystal structure of the Bacteroides thetaiotaomicron protein BT_3984 was determined to a resolution of 1.7 Å and was the first structure to be determined from the extensive SusD family of polysaccharide-binding proteins. SusD is an essential component of the sus operon that defines the paradigm for glycan utilization in dominant members of the human gut microbiota. Structural analysis of BT_3984 revealed an N-terminal region containing several tetratricopeptide repeats (TPRs), while the signature C-terminal region is less structured and contains extensive loop regions. Sequence and structure analysis of BT_3984 suggests the presence of binding interfaces for other proteins from the polysaccharide-utilization complex.

The crystal structures of SPO0140 and Sbal_2486 were determined using the semiautomated high-throughput pipeline of the Joint Center for Structural Genomics (JCSG) as part of the NIGMS Protein Structure Initiative (PSI). The structures revealed a conserved core with domain duplication and a superficial similarity of the C-terminal domain to pleckstrin homology-like folds. The conservation of the domain interface indicates a potential binding site that is likely to involve a nucleotide-based ligand, with genome-context and gene-fusion analyses additionally supporting a role for this family in signal transduction, possibly during oxidative stress.

KPN03535 (gi|152972051) is a putative lipoprotein of unknown function that is secreted by Klebsiella pneumoniae MGH 78578. The crystal structure reveals that despite a lack of any detectable sequence similarity to known structures, it is a novel variant of the OB-fold and structurally similar to the bacterial Cpx-pathway protein NlpE, single-stranded DNA-binding (SSB) proteins and toxins. K. pneumoniae MGH 78578 forms part of the normal human skin, mouth and gut flora and is an opportunistic pathogen that is linked to about 8% of all hospital-acquired infections in the USA. This structure provides the foundation for further investigations into this divergent member of the OB-fold family.

Membrane-attack complex/perforin (MACPF) proteins are transmembrane pore-forming proteins that are important in both human immunity and the virulence of pathogens. Bacterial MACPFs are found in diverse bacterial species, including most human gut-associated Bacteroides species. The crystal structure of a bacterial MACPF-domain-containing protein BT_3439 (Bth-MACPF) from B. thetaiotaomicron, a predominant member of the mammalian intestinal microbiota, has been determined. Bth-MACPF contains a membrane-attack complex/perforin (MACPF) domain and two novel C-terminal domains that resemble ribonuclease H and interleukin 8, respectively. The entire protein adopts a flat crescent shape, characteristic of other MACPF proteins, that may be important for oligomerization. This Bth-MACPF structure provides new features and insights not observed in two previous MACPF structures. Genomic context analysis infers that Bth-MACPF may be involved in a novel protein-transport or nutrient-uptake system, suggesting an important role for these MACPF proteins, which were likely to have been inherited from eukaryotes via horizontal gene transfer, in the adaptation of commensal bacteria to the host environment.

Sufu (Suppressor of Fused), a two-domain protein, plays a critical role in regulating Hedgehog signaling and is conserved from flies to humans. A few bacterial Sufu-like proteins have previously been identified based on sequence similarity to the N-terminal domain of eukaryotic Sufu proteins, but none have been structurally or biochemically characterized and their function in bacteria is unknown. We have determined the crystal structure of a more distantly related Sufu-like homolog, NGO1391 from Neisseria gonorrhoeae, at 1.4 Å resolution, which provides the first biophysical characterization of a bacterial Sufu-like protein. The structure revealed a striking similarity to the N-terminal domain of human Sufu (r.m.s.d. of 2.6 Å over 93% of the NGO1391 protein), despite an extremely low sequence identity of ∼15%. Subsequent sequence analysis revealed that NGO1391 defines a new subset of smaller, Sufu-like proteins that are present in ∼200 bacterial species and has resulted in expansion of the SUFU (PF05076) family in Pfam.

The crystal structure of the Bacteroides thetaiotaomicron protein BT_3984 was determined to a resolution of 1.7 Å and was the first structure to be determined from the extensive SusD family of polysaccharide-binding proteins. SusD is an essential component of the sus operon that defines the paradigm for glycan utilization in dominant members of the human gut microbiota. Structural analysis of BT_3984 revealed an N-terminal region containing several tetratricopeptide repeats (TPRs), while the signature C-terminal region is less structured and contains extensive loop regions. Sequence and structure analysis of BT_3984 suggests the presence of binding interfaces for other proteins from the polysaccharide-utilization complex.

KPN03535 (gi|152972051) is a putative lipoprotein of unknown function that is secreted by Klebsiella pneumoniae MGH 78578. The crystal structure reveals that despite a lack of any detectable sequence similarity to known structures, it is a novel variant of the OB-fold and structurally similar to the bacterial Cpx-pathway protein NlpE, single-stranded DNA-binding (SSB) proteins and toxins. K. pneumoniae MGH 78578 forms part of the normal human skin, mouth and gut flora and is an opportunistic pathogen that is linked to about 8% of all hospital-acquired infections in the USA. This structure provides the foundation for further investigations into this divergent member of the OB-fold family.

BT1062 from Bacteroides thetaiotaomicron is a homolog of Mfa2 (PGN0288 or PG0179), which is a component of the minor fimbriae in Porphyromonas gingivalis. The crystal structure of BT1062 revealed a conserved fold that is widely adopted by fimbrial components.