Sagittal craniosynostosis, the most common form of craniosynostosis, affects 1 per 1000 live births. The main surgical treatments include endoscopic suturectomy and open cranial vault remodeling. ...This video describes an open reconstruction method, including strip resection of the sagittal suture, biparietal craniotomies with spiral cut cranioplasty, and barrel staves of the posterior occiput. Ideally used between 4 and 15 months of age, this approach takes advantage of the flexibility of the cranial bones to expand, allowing for immediate and long-term increases of the parietal width and correction of cosmetic deformity, without necessitating the use of cranial molding devices postoperatively. The video can be found here: https://vimeo.com/516699203
The B7 family plays a critical role in both positive and negative regulation of immune responses by engaging a variety of receptors on lymphocytes. Importantly, blocking coinhibitory molecules using ...antibodies specific for CTLA-4 and PD-1 enhances tumor immunity in a subset of patients. Therefore, it is critical to understand the role of different B7 family members since they may be suitable therapeutic targets. B7-H4 is another member that inhibits T-cell function, and it is also upregulated on a variety of tumors and has been proposed to promote tumor growth. Here, we investigate the role of B7-H4 in tumor development and show that B7-H4 expression inhibits tumor growth in two mouse models. Furthermore, we show that B7-H4 expression is required for antitumor immune responses in a mouse model of mammary tumorigenesis. We found that the expression levels of B7-H4 correlate with MHC class I expression in both mouse and human samples. We show that IFNγ upregulates B7-H4 expression on mouse embryo fibroblasts and that the upregulation of B7-H4 on tumors is dependent on T cells. Notably, patients with breast cancer with increased B7-H4 expression show a prolonged time to recurrence. These studies demonstrate a positive role for B7-H4 in promoting antitumor immunity.
A method for the determination of a prostaglandin D(2) receptor antagonist (I, a compound being evaluated for the prevention of niacin induced flushing) and its acyl glucuronide metabolite (II) in ...human plasma is presented. The method utilized high performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection using an atmospheric pressure chemical ionization (APCI) interface operated in the positive ionization mode. The product ion was a radical cation generated via a homolytic bond cleavage. A chemical analog of the drug was used as internal standard (III). The acyl glucuronide metabolite (II) was detected using the same precursor-to-product ion transition used for the parent compound after chromatographic separation of I and II. Drug and metabolite were extracted using semi-automated, 96-well format solid phase extraction (SPE), and chromatography was performed using a reverse phase analytical column with an isocratic mobile phase. The chromatographic retention factor (k') of II was found to be highly sensitive to mobile phase formic acid concentration. An adjustment in mobile phase formic acid concentration improved the chromatographic separation between II and a mono-hydroxylated metabolite after an unexpected lack of MS/MS selectivity between the two molecules was observed. The dependence of retention factor on formic acid concentration (k' increased as formic acid concentration decreased) was thought to indicate polar interactions between II and the stationary phase. The stability of II in spiked human plasma was determined. The rate of hydrolysis back to parent compound was relatively low (approximately 0.1 and 0.5% per hour at room temperature and 4 degrees C, respectively) indicating that significant changes in analyte concentrations did not occur during sample processing. The concentration range of the assay was 10-2500 ng/mL for both drug and glucuronide metabolite.
A method for the determination of a prostaglandin D
2 receptor antagonist (
I, a compound being evaluated for the prevention of niacin induced flushing) and its acyl glucuronide metabolite (
II) in ...human plasma is presented. The method utilized high performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection using an atmospheric pressure chemical ionization (APCI) interface operated in the positive ionization mode. The product ion was a radical cation generated via a homolytic bond cleavage. A chemical analog of the drug was used as internal standard (
III). The acyl glucuronide metabolite (
II) was detected using the same precursor-to-product ion transition used for the parent compound after chromatographic separation of
I and
II. Drug and metabolite were extracted using semi-automated, 96-well format solid phase extraction (SPE), and chromatography was performed using a reverse phase analytical column with an isocratic mobile phase. The chromatographic retention factor (
k′) of
II was found to be highly sensitive to mobile phase formic acid concentration. An adjustment in mobile phase formic acid concentration improved the chromatographic separation between
II and a mono-hydroxylated metabolite after an unexpected lack of MS/MS selectivity between the two molecules was observed. The dependence of retention factor on formic acid concentration (
k′ increased as formic acid concentration decreased) was thought to indicate polar interactions between
II and the stationary phase. The stability of
II in spiked human plasma was determined. The rate of hydrolysis back to parent compound was relatively low (approximately 0.1 and 0.5% per hour at room temperature and 4
°C, respectively) indicating that significant changes in analyte concentrations did not occur during sample processing. The concentration range of the assay was 10–2500
ng/mL for both drug and glucuronide metabolite.
We modified and adapted a novel noncontact speckle contrast diffuse correlation tomography (scDCT) system, developed in our laboratory, for noninvasive 3D imaging of blood flow distributions in small ...animals and large human tissue volumes. The new system has the flexibility of selecting small and large regions of interest (ROIs) with different imaging depths for the use in both the small mouse head and large human mastectomy skin flaps. The capability of the upgraded scDCT system was demonstrated by continuous imaging of cerebral blood flow (CBF) changes in a small mouse head during sequential common carotid artery ligations and blood flow distributions in human mastectomy skin flaps during surgery. As expected, arterial occlusions resulted in substantial CBF decreases in deep brain tissues of the mouse. Spatial heterogeneity existed, and lower blood flow values were observed around the incision area of mastectomy skin flaps. The noncontact scDCT system with flexible ROIs and imaging depths has the potential to be used in many preclinical and clinical settings for noninvasive assessment of tissue hemodynamics and viability.
The emerging SARS-CoV-2 variants of concern (VOCs) threaten the effectiveness of current COVID-19 vaccines administered intramuscularly and designed to only target the spike protein. There is a ...pressing need to develop next-generation vaccine strategies for broader and long-lasting protection. Using adenoviral vectors (Ad) of human and chimpanzee origin, we evaluated Ad-vectored trivalent COVID-19 vaccines expressing spike-1, nucleocapsid, and RdRp antigens in murine models. We show that single-dose intranasal immunization, particularly with chimpanzee Ad-vectored vaccine, is superior to intramuscular immunization in induction of the tripartite protective immunity consisting of local and systemic antibody responses, mucosal tissue-resident memory T cells and mucosal trained innate immunity. We further show that intranasal immunization provides protection against both the ancestral SARS-CoV-2 and two VOC, B.1.1.7 and B.1.351. Our findings indicate that respiratory mucosal delivery of Ad-vectored multivalent vaccine represents an effective next-generation COVID-19 vaccine strategy to induce all-around mucosal immunity against current and future VOC.
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•Two trivalent adenoviral-vectored COVID-19 vaccines were developed and evaluated•Intranasal, but not intramuscular, immunization induces tripartite mucosal immunity•Intranasal immunization protects against ancestral and variant strains of SARS-CoV-2•Optimal protection requires B and T cell immunity and trained innate immunity
Respiratory mucosal immunization with a next-generation adenoviral-vectored trivalent COVID-19 vaccine expressing spike, nucleocapsid, and RdRp antigens, induces all-around protective mucosal immunity against SARS-CoV-2 via induction of systemic and local antibodies, lung-tissue-resident memory T cells, and trained alveolar macrophages.
ObjectiveThe degree awarding gap indicates that racially minoritised higher education students receive lower degree classifications relative to their white peers. While the reasons for this are ...complex, research suggests that educator and practitioner attitudes and behaviour towards racially minoritised students are a significant contributing factor. This preregistered study evaluates the effectiveness of unconscious racial bias training (URBT) to enhance National Health Service senior practitioner’s recognition of how racial inequalities negatively impact racially minoritised students.DesignA mixed-methods study with a pretest and post-test design was conducted in the higher education and healthcare practice environment.MethodsForty-nine NHS senior practitioners completed a 4-hour URBT workshop with activities focusing on activating stereotypes, exploring differences between unconscious and implicit bias, discussing the development of bias, and reflecting on student experiences of prejudice, harassment and discrimination. They completed pre- and post- quantitative measures that assessed the effectiveness of URBT and changes in racial competency, awareness and perceptions of unconscious racial bias. Qualitative measures explored the usefulness and perceived applications of URBT, and a 1-month follow-up assessed further how it had been applied within practice.ResultsParticipants reported positive evaluations of URBT, higher perceived racial competency, awareness and perceptions of racial bias (ps<0.001, dz>0.35). After 1 month, key themes from qualitative responses suggested that participants had increased self-awareness and were exploring how to set up mentoring and working groups, change recruitment and progression processes, and diversify the taught curriculum.ConclusionsURBT may be one effective strategy to enhance awareness and encourage reflections of racial bias. We discuss how reducing racial inequalities requires a multifaceted approach that affords upfront conversations about systemic racism, implements effective initiatives, policies and procedures, and engages in continuous evaluation.
This study explored the lived experiences of racial bias for Black, Asian and Minority Ethnic students undertaking an undergraduate or post-graduate degree in nursing, midwifery and allied health ...courses in the United Kingdom.
Previous research indicates that students from Black, Asian and Minority Ethnic groups have fewer opportunities to succeed at university and this has brought about a race awarding gap in their degree attainment. The reasons for this awarding gap are complex and multi-factorial and it is crucial that the lived experiences of racial bias are explored from the student perspective.
A hermeneutic phenomenological approach was adopted to elicit individual and collective experiences in the practice environment, a mandatory component of the student’s degree.
A focus group and individual semi-structured interviews were conducted to collect data from sixteen participants and analysed using thematic analysis
Three encompassing themes were identified which included a sense of not belonging, trauma impact on mental health and understanding covert and overt racism. Participants reported incidences of racism and appeared to be traumatised by their experiences within practice and the university. They also reported poor mental health and well-being as shared experiences and a lack of confidence in the university and practice to mitigate racial issues.
Meaningful action must be taken by universities and practice partners to advance racial inequality initiatives by having robust anti-racism action plans and processes. These should be co-created with students and staff to reduce the race awarding gap.
Vaccination route dictates the quality and localization of immune responses within tissues. Intranasal vaccination seeds tissue-resident adaptive immunity, alongside trained innate responses within ...the lung/airways, critical for superior protection against SARS-CoV-2. This protocol encompasses intranasal vaccination in mice, step-by-step bronchoalveolar lavage for both cellular and acellular airway components, lung mononuclear cell isolation, and detailed flow cytometric characterization of lung tissue-resident memory T cell responses, and airway macrophage-trained innate immunity.
For complete details on the use and execution of this protocol, please refer to Afkhami et al. (2022).
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•Step-by-step procedure for intranasal vaccination in mice•Protocol for bronchoalveolar lavage and isolating airway cellular/acellular components•Detailed protocol for processing and isolation of lung mononuclear cells•Flow cytometry protocol for lung tissue-resident innate/adaptive memory responses
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Vaccination route dictates the quality and localization of immune responses within tissues. Intranasal vaccination seeds tissue-resident adaptive immunity, alongside trained innate responses within the lung/airways, critical for superior protection against SARS-CoV-2. This protocol encompasses intranasal vaccination in mice, step-by-step bronchoalveolar lavage for both cellular and acellular airway components, lung mononuclear cell isolation, and detailed flow cytometric characterization of lung tissue-resident memory T cell responses, and airway macrophage-trained innate immunity.
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