A VHH single-domain platform enabling discovery and development of monospecific antibodies and modular neutralizing bispecifics against SARS-CoV-2 variants.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to evolve, escape coronavirus disease 2019 therapeutics and vaccines, and jeopardize public health. To combat SARS-CoV-2 antigenic escape, we developed a rapid, high-throughput pipeline to discover monospecific VHH antibodies and iteratively develop VHH-Fc-VHH bispecifics capable of neutralizing emerging SARS-CoV-2 variants. By panning VHH single-domain phage libraries against ancestral or beta spike proteins, we discovered high-affinity VHH antibodies with unique target epitopes. Combining two VHHs into a tetravalent bispecific construct conferred broad neutralization activity against multiple variants and was more resistant to antigenic escape than the monospecific antibody alone. Following the rise of the Omicron variant, a VHH in the original bispecific construct was replaced with another VHH discovered against the Omicron BA.1 receptor binding domain; the resulting bispecific exhibited neutralization against both BA.1 and BA.5 sublineage variants. A heavy chain-only tetravalent VHH-Fc-VHH bispecific platform derived from humanized synthetic libraries held a myriad of unique advantages: (i) synthetic preconstructed libraries minimized risk of liabilities and maximized discovery speed, (ii) VHH scaffolds allowed for a modular "plug-and-play" format that could be rapidly iterated upon as variants of concern arose, (iii) natural dimerization of single VHH-Fc-VHH polypeptides allowed for straightforward bispecific production and purification methods, and (iv) multivalent approaches enhanced avidity boosting effects and neutralization potency, and conferred more robust resistance to antigenic escape than monovalent approaches against specific variants. This iterative platform of rapid VHH discovery combined with modular bispecific design holds promise for long-term viral control efforts.

Discovery of a potent, selective, and tumor-suppressing antibody antagonist of adenosine A2A receptor.

New immune checkpoints are emerging in a bid to improve response rates to immunotherapeutic drugs. The adenosine A2A receptor (A2AR) has been proposed as a target for immunotherapeutic development due to its participation in immunosuppression of the tumor microenvironment. Blockade of A2AR could restore tumor immunity and, consequently, improve patient outcomes. Here, we describe the discovery of a potent, selective, and tumor-suppressing antibody antagonist of human A2AR (hA2AR) by phage display. We constructed and screened four single-chain variable fragment (scFv) libraries-two synthetic and two immunized-against hA2AR and antagonist-stabilized hA2AR. After biopanning and ELISA screening, scFv hits were reformatted to human IgG and triaged in a series of cellular binding and functional assays to identify a lead candidate. Lead candidate TB206-001 displayed nanomolar binding of hA2AR-overexpressing HEK293 cells; cross-reactivity with mouse and cynomolgus A2AR but not human A1, A2B, or A3 receptors; functional antagonism of hA2AR in hA2AR-overexpressing HEK293 cells and peripheral blood mononuclear cells (PBMCs); and tumor-suppressing activity in colon tumor-bearing HuCD34-NCG mice. Given its therapeutic properties, TB206-001 is a good candidate for incorporation into next-generation bispecific immunotherapeutics.

"They were quick, insipid, and stuck to the typical medical checkups": A Narrative Study on Women's Expectations and Experiences of Maternity Care in Bangladesh.

The quality of healthcare service delivery is generally determined by how patients' expectations were met successfully. This narrative study explores how women perceive and experience healthcare during childbirth in the context of Bangladesh. This study is inspired by Clandinin and Connelly's three-dimensional paradigm of narrativity that combines temporality, social interaction, and place. To unearth patient-driven narratives, the researcher purposely picked 12 women who gave birth in different private and public health facilities in Bangladesh. Four themes standout from the women's narratives. Excerpts of women's stories have been included in discussing the themes as well as author's conviction on this phenomenon. Most of the participants experienced a shared level of difficulty in choosing the health facilities (private vs public), motivated primarily by delivery costs and social background. Women with a higher level of education and financial means often opted to give birth in private facilities due to their negative perception and experience of the public facility. There was evident discontent when doctors decided for cesarean deliveries. Women were dissatisfied by providers' general lack of empathy and vicarious emotion. However, those women who gave birth in public hospitals expressed some degree of satisfaction which might be attributed to their low expectations and moderate social standing. Women's stories also delved into how societal norms, taboos, and elderly relatives put them in uncomfortable situations. To improve patient-provider interactions, healthcare practitioners should prioritize patient-centered care and collaborative decision-making. Reducing healthcare disparity and resolving superannuated pregnancy norms are also critical challenges.

Menopausal experiences of women of Chinese ethnicity: A meta-ethnography.

OBJECTIVE: Menopause and the changes it brings to a woman's life necessitate a comprehensive approach to face and experience the transition. This paper aims at synthesizing results from qualitative studies of menopausal experiences among Chinese and other women of similar ethnicity and culture. DESIGN AND METHOD: A comprehensive search strategy of multiple databases along with bibliographic hand searches was employed to identify qualitative studies published in English peer-reviewed journals between 2008 and 2022 focused on the menopausal experiences (peri and post-menopause) of Chinese and other women of similar ethnic backgrounds. Twelve studies met the inclusion criteria. The final sample consisted of 238 women aged between 40 to 60 years who had experienced menopausal symptoms. This qualitative systematic literature review adopted Noblit and Hare's seven-stage theoretical meta-ethnographic approach to construct an inductive and interpretive form of synthesis and subsequent analysis. SYNTHESES OF FINDINGS: The synthesis of primary data identified four key concepts that entitle women's menopausal experiences: being menopausal, ramifications on well-being, family and social support around menopause, and healthcare throughout menopause. The subsequent second-order interpretation revealed that women accepted the inevitability of the natural aging process in the decline of sexual drive, reinvented relationships with partners, and expressed the significance of a supportive environment in order to successfully navigate the transition. Third-order interpretations sought to establish a link between physiological complications, loss of femininity, patriarchal-dominated societal norms, and a support system that spans the entire menopause experience. Healthcare's contribution has also been deemed to be insufficient due to a lack of information and empathy from health experts. Negligence or reluctance to seek healthcare and skepticism toward hormone replacement therapy (HRT) had also been a source of concern, as they have had the potential to exacerbate medical difficulties and emotional turmoil. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: A comprehensive approach that considers women's physiological and psychological well-being and major attempts to change cultural beliefs and norms about women's sexual health may be effective in aiding menopausal women during their transition. Additionally, appropriate guidelines and management should be in place to enable women to address menopause difficulties effectively with the assistance of healthcare experts and the support of their families and community.

Optimization of a Glucagon-Like Peptide 1 Receptor Antagonist Antibody for Treatment of Hyperinsulinism.

Congenital hyperinsulinism (HI) is a genetic disorder in which pancreatic ß-cell insulin secretion is excessive and results in hypoglycemia that, without treatment, can cause brain damage or death. Most patients with loss-of-function mutations in ABCC8 and KCNJ11, the genes encoding the ß-cell ATP-sensitive potassium channel (KATP), are unresponsive to diazoxide, the only U.S. Food and Drug Administration-approved medical therapy and require pancreatectomy. The glucagon-like peptide 1 receptor (GLP-1R) antagonist exendin-(9-39) is an effective therapeutic agent that inhibits insulin secretion in both HI and acquired hyperinsulinism. Previously, we identified a highly potent antagonist antibody, TB-001-003, which was derived from our synthetic antibody libraries that were designed to target G protein-coupled receptors. Here, we designed a combinatorial variant antibody library to optimize the activity of TB-001-003 against GLP-1R and performed phage display on cells overexpressing GLP-1R. One antagonist, TB-222-023, is more potent than exendin-(9-39), also known as avexitide. TB-222-023 effectively decreased insulin secretion in primary isolated pancreatic islets from a mouse model of hyperinsulinism, Sur1-/- mice, and in islets from an infant with HI, and increased plasma glucose levels and decreased the insulin to glucose ratio in Sur1-/- mice. These findings demonstrate that targeting GLP-1R with an antibody antagonist is an effective and innovative strategy for treatment of hyperinsulinism. ARTICLE HIGHLIGHTS: Patients with the most common and severe form of diazoxide-unresponsive congenital hyperinsulinism (HI) require a pancreatectomy. Other second-line therapies are limited in their use because of severe side effects and short half-lives. Therefore, there is a critical need for better therapies. Studies with the glucagon-like peptide 1 receptor (GLP-1R) antagonist, avexitide (exendin-(9-39)), have demonstrated that GLP-1R antagonism is effective at lowering insulin secretion and increasing plasma glucose levels. We have optimized a GLP-1R antagonist antibody with more potent blocking of GLP-1R than avexitide. This antibody therapy is a potential novel and effective treatment for HI.

In vitro assessment of the genotoxicity and immunotoxicity of treated and untreated municipal effluents and receiving waters in freshwater organisms.

Municipal wastewater effluent is one of the largest sources of pollution entering surface waters in the Laurentian Great Lakes. Exposure to wastewater effluent has been associated with impaired immune systems and induction of genotoxicity to aquatic animals. Due to habitat degradation and environmental pollution linked to industrial development and population growth, several regions of the Great Lakes have been designated Areas of Concern (AOCs). In this study, we assessed the effect of extracts of sewage influent, (treated) effluent and receiving surface waters from the Hamilton Harbour AOC and the Toronto and Region AOC (Ontario, Canada) on the phagocytic immune response of rainbow trout (Oncorhynchus mykiss) kidney leukocytes and the genotoxicity (DNA strand breaks) of these extracts on freshwater mussel (Eurynia dilatata) hemocytes. We identified and quantified numerous chemicals present in the various samples extracted for exposure. In freshwater mussels, extracts from Hamilton Harbour AOC induced DNA damage with the most frequency (12 out of 28 samples) regardless of sample type, reflecting past and present industrial activities. In contrast, extracts from Toronto and Region AOC induced DNA damage infrequently (2 out of 32 (summer) and 5 out of 32 (fall) samples, respectively) and from different WWTPs at different times. None of the extracts induced any significant effect on phagocytosis of rainbow trout kidney leukocytes. The present study indicates that despite overall improvements to effluent quality, treatment of influent by WWTPs may not result in a corresponding improvement of the genotoxicity of effluents. In vitro bioassays are useful and cost-effective rapid-screening tools for preliminary assessments of contamination of aquatic ecosystems.

"Do we even have a voice?" Health providers' perspective on the patient accommodation strategies in Bangladesh.

OBJECTIVE: In a resource-constrained setting like Bangladesh, effective patient-provider communication is critical to the delivery of maternal healthcare services. Using communication accommodation theory, this study tried to understand how providers perceive, engage, and accommodate patients' needs in maternity wards. METHODS: This qualitative exploratory study used a semi-structured interview guide to conduct in-depth face-to-face interviews of ten healthcare providers in two government-funded public health facilities in Bangladesh. The interview data were analyzed using MAXQDA 2020 software. RESULTS: The thematic analyses revealed that nurses and midwives faced conceivable neglect from patients and their attendees due to possible service and facility constraints, indicating their low status and control within the service operation. Despite efforts to address patients' emotional and psychological concerns, providers appear to avoid (divergence strategy) confronting patients and their irate visitors. Unimodal convergence emerged when providers accepted patients' arguments about the systematic inadequacy of service facilities. Providers have employed interpretability tactics to communicate medical opinions and applied nonverbal cues where necessary. CONCLUSION: A lack of open communication between healthcare providers and patients necessitated various forms of accommodation. Integrated strategies addressing service restrictions and initiatives fostering patient understanding and cooperation may improve patient-provider communication.

DNA polymerase θ promotes CAG•CTG repeat expansions in Huntington's disease via insertion sequences of its catalytic domain.

Huntington's disease (HD), a neurodegenerative disease characterized by progressive dementia, psychiatric problems, and chorea, is known to be caused by CAG repeat expansions in the HD gene HTT. However, the mechanism of this pathology is not fully understood. The translesion DNA polymerase θ (Polθ) carries a large insertion sequence in its catalytic domain, which has been shown to allow DNA loop-outs in the primer strand. As a result of high levels of oxidative DNA damage in neural cells and Polθ's subsequent involvement in base excision repair of oxidative DNA damage, we hypothesized that Polθ contributes to CAG repeat expansion while repairing oxidative damage within HTT. Here, we performed Polθ-catalyzed in vitro DNA synthesis using various CAG•CTG repeat DNA substrates that are similar to base excision repair intermediates. We show that Polθ efficiently extends (CAG)n•(CTG)n hairpin primers, resulting in hairpin retention and repeat expansion. Polθ also triggers repeat expansions to pass the threshold for HD when the DNA template contains 35 repeats upward. Strikingly, Polθ depleted of the catalytic insertion fails to induce repeat expansions regardless of primers and templates used, indicating that the insertion sequence is responsible for Polθ's error-causing activity. In addition, the level of chromatin-bound Polθ in HD cells is significantly higher than in non-HD cells and exactly correlates with the degree of CAG repeat expansion, implying Polθ's involvement in triplet repeat instability. Therefore, we have identified Polθ as a potent factor that promotes CAG•CTG repeat expansions in HD and other neurodegenerative disorders.

MIF is a 3' flap nuclease that facilitates DNA replication and promotes tumor growth.

How cancer cells cope with high levels of replication stress during rapid proliferation is currently unclear. Here, we show that macrophage migration inhibitory factor (MIF) is a 3' flap nuclease that translocates to the nucleus in S phase. Poly(ADP-ribose) polymerase 1 co-localizes with MIF to the DNA replication fork, where MIF nuclease activity is required to resolve replication stress and facilitates tumor growth. MIF loss in cancer cells leads to mutation frequency increases, cell cycle delays and DNA synthesis and cell growth inhibition, which can be rescued by restoring MIF, but not nuclease-deficient MIF mutant. MIF is significantly upregulated in breast tumors and correlates with poor overall survival in patients. We propose that MIF is a unique 3' nuclease, excises flaps at the immediate 3' end during DNA synthesis and favors cancer cells evading replication stress-induced threat for their growth.

A Case of Dubin-Johnson Syndrome Presenting as Neonatal Cholestasis With Paucity of Interlobular Bile Ducts.

Dubin-Johnson syndrome (DJS) is a rare autosomal recessive disorder that typically manifests in young adulthood as jaundice with conjugated hyperbilirubinemia. We report a case presenting as neonatal cholestasis with the unexpected histologic finding of paucity of interlobular bile ducts, a feature that is not typically seen in DJS. The diagnosis was confirmed by absent canalicular multidrug-resistance-associated protein 2 (MRP2) immunohistochemical staining on liver biopsy tissue and molecular genetic testing that demonstrated heterozygous mutations in the ATP-Binding Cassette Subfamily C Member 2 (ABCC2) gene, including a novel missense mutation. This report describes a case of DJS with atypical clinicopathologic findings and suggests that DJS should be considered in patients with neonatal cholestasis and bile duct paucity.

Experience of Stress and Burnout among Pastors in China.

A quantitative survey was conducted among a convenience sample of 164 church ministers in China to test a model relating experience of stress, clergy burnout, and clergy health. Results indicated that respondents experience relatively low levels of stress and burnout. The burnout score was 2.73 on a seven-point scale. Stress from family was a significant predictor of overall burnout. The experience of emotional exhaustion was a significant predictor of the occurrence of adverse health symptoms.

Adolescents from low-income families in Hong Kong and unhealthy eating behaviours: Implications for health and social care practitioners.

The development of dietary preferences of adolescents involves a complex interplay of individual behaviours and environmental factors. Interpersonal factors-such as peer influences and unpleasant school experiences-and institutional factors-such as school rules and policies-are closely associated with unhealthy eating of adolescents. Family support and guidance are also crucial in influencing adolescents' eating habits. However, the low social status, low educational levels, and low household incomes of disadvantaged parents can markedly prevent their children from establishing healthy eating habits. Therefore, adolescents from low-income families are more likely to engage in unhealthy dietary behaviours and hence to be more susceptible to diet-related health problems. However, few studies have addressed the difficulties associated with inculcating healthy eating habits among adolescents from low-income families. Therefore, to investigate the barriers to adopting healthy eating habits, this study adopted a qualitative research approach and conducted five focus-group semistructured interviews with 30 junior- and senior-form students of a secondary school in Hong Kong, all of whom were from low-income families. The results revealed skipping meals because of poverty, following irregular meal patterns on school holidays, receiving poor guidance from family and peers, perceiving healthy eating as expensive and unappealing, and geographical inaccessibility to healthy food all prevented these students from healthy eating. These mutually reinforcing factors were interlocking with the economic strain that was experienced by the participants and their families. In particular, the stereotype of "healthful food is expensive" was strong. Therefore, we suggest students from low-income families should be enabled to understand that healthy eating is not necessarily expensive. The participants' stereotypes about healthy food was handed down by their parents. Such stereotypes, together with the low health literacy, influence the food preparation habits of the parents. Therefore, parents should be made to aware that healthful food can also be affordable.

Experience of Stress and Coping Strategies among Pastors' Wives in China.

A qualitative study was conducted to examine the experience of stress and coping strategies among 15 pastors' wives from a city in mainland China. Results indicated that nearly all interviewees experienced financial stress and loneliness, a phenomenon consistent with that seen in literature in the West. However, stress arising from role expectations among the spouses' congregations was low. Most interviewees coped with stress through family and social support, as well as through praying.

MutSß abundance and Msh3 ATP hydrolysis activity are important drivers of CTG•CAG repeat expansions.

CTG•CAG repeat expansions cause at least twelve inherited neurological diseases. Expansions require the presence, not the absence, of the mismatch repair protein MutSß (Msh2-Msh3 heterodimer). To evaluate properties of MutSß that drive expansions, previous studies have tested under-expression, ATPase function or polymorphic variants of Msh2 and Msh3, but in disparate experimental systems. Additionally, some variants destabilize MutSß, potentially masking the effects of biochemical alterations of the variations. Here, human Msh3 was mutated to selectively inactivate MutSß. Msh3-/- cells are severely defective for CTG•CAG repeat expansions but show full activity on contractions. Msh3-/- cells provide a single, isogenic system to add back Msh3 and test key biochemical features of MutSß on expansions. Msh3 overexpression led to high expansion activity and elevated levels of MutSß complex, indicating that MutSß abundance drives expansions. An ATPase-defective Msh3 expressed at normal levels was as defective in expansions as Msh3-/- cells, indicating that Msh3 ATPase function is critical for expansions. Expression of two Msh3 polymorphic variants at normal levels showed no detectable change in expansions, suggesting these polymorphisms primarily affect Msh3 protein stability, not activity. In summary, CTG•CAG expansions are limited by the abundance of MutSß and rely heavily on Msh3 ATPase function.

Perception of acupuncture among users and nonusers: A qualitative study.

This study uses a qualitative methodology to examine the perception of acupuncture among users and nonusers. Altogether 37 participants, age 35 or older, were interviewed. Participants' perception of advantages and disadvantages of adopting acupuncture, and their criteria in selecting acupuncturists, were collected. Results found that among the user group, acupuncture was perceived as being effective, having little side effects, and generating lasting impact. Among nonusers, acupuncture was perceived as lacking a clinical base, high risk, and nonstandardized. Nonusers had less confidence in acupuncture than biomedicine. Participants relied on social communication and the practitioner's professional qualifications in choosing acupuncturists. Marketing implications are discussed.

The social and community opportunities profile social inclusion measure: Structural equivalence and differential item functioning in community mental health residents in Hong Kong and the United Kingdom.

INTRODUCTION: China's future major health problem will be the management of chronic diseases - of which mental health is a major one. An instrument is needed to measure mental health inclusion outcomes for mental health services in Hong Kong and mainland China as they strive to promote a more inclusive society for their citizens and particular disadvantaged groups. AIM: To report on the analysis of structural equivalence and item differentiation in two mentally unhealthy and one healthy sample in the United Kingdom and Hong Kong. METHOD: The mental health sample in Hong Kong was made up of non-governmental organisation (NGO) referrals meeting the selection/exclusion criteria (being well enough to be interviewed, having a formal psychiatric diagnosis and living in the community). A similar sample in the United Kingdom meeting the same selection criteria was obtained from a community mental health organisation, equivalent to the NGOs in Hong Kong. Exploratory factor analysis and logistic regression were conducted. RESULTS: The single-variable, self-rated 'overall social inclusion' differs significantly between all of the samples, in the way we would expect from previous research, with the healthy population feeling more included than the serious mental illness (SMI) groups. In the exploratory factor analysis, the first two factors explain between a third and half of the variance, and the single variable which enters into all the analyses in the first factor is having friends to visit the home. All the regression models were significant; however, in Hong Kong sample, only one-fifth of the total variance is explained. CONCLUSION: The structural findings imply that the social and community opportunities profile-Chinese version (SCOPE-C) gives similar results when applied to another culture. As only one-fifth of the variance of 'overall inclusion' was explained in the Hong Kong sample, it may be that the instrument needs to be refined using different or additional items within the structural domains of inclusion.

Application of SCOPE-C to Measure Social Inclusion Among Mental Health Services Users in Hong Kong.

This study describes the construction of the Chinese version of the Social and Communities Opportunities Profile (SCOPE), henceforth, the SCOPE-C, to measure social inclusion among mental health services users in Hong Kong. The SCOPE-C was developed based on concept-mapping and benchmarking of census questions. The questionnaire consisted of 56 items, went through a standardized linguistic validation process and was pilot tested with qualitative feedback from five users of mental health services. Altogether 168 Chinese service users were recruited through various NGO mental health services to have three times face-to-face interview between October 2013 and July 2014. Results indicated that items related to satisfaction with opportunities and perceived opportunities in various social domains had high consistency. Nearly all the Kappa statistics and Pearson correlation coefficients between the baseline and two rounds of re-test were significant. The SCOPE-C was considered a valid instrument for Hong Kong mental health user population.

Benefits and stressors - Perceived effects of ICT use on employee health and work stress: An exploratory study from Austria and Hong Kong.

Stress has become a mass phenomenon in the modern workplace. The use of information and communication technologies is beginning to receive greater attention in the context of occupational stress. An exploratory qualitative study was conducted to examine both stressors and benefits resulting from technologies among practitioners in the advertising, public relations, and journalism industry in Hong Kong and Austria. Results suggest that technologies allow instant availability, which facilitates communication processes as well as information exchange. Notably, modern technologies enable employees to organize their work with greater temporal and spatial flexibility, thus creating an opportunity for better balancing work and private life. However, evolving technologies have come with a cost; the pressure to be constantly available via technologies constitutes a major source of stress, increasing the risk of experiencing prolonged work stress and its adverse consequences on employee health and well-being, such as a burnout. Furthermore, findings suggest that availability pressure may be attributed to an inner obligation rather than to an organizational expectation. Hence, making employees aware of their connectivity behaviour may help to diminish the experience of technology-induced work stress and improve and maintain employees' health and well-being in the long term. Practical implications, limitations, and future research directions are provided.

Should different marketing communication strategies be used to promote healthy eating among male and female adolescents?

A study was conducted to examine how interpersonal norms, media norms, attitudes, perceived behavioral control, perceived barriers, and self-efficacy had an influence on healthy eating intention among adolescents. A probability sample of 544 adolescents aged 12 to 18 was conducted. Results indicated that girls had a more favorable attitude and intention toward healthy eating than boys. Healthy eating intention among boys was predicted by attitude, perceived behavioral control, perceived barriers, and self-efficacy, and among girls was predicted by perceived behavioral control and self-efficacy. Different marketing strategies to promote healthy eating among adolescent boys and girls should be adopted.

Gains and losses in creative personality as perceived by adults across the life span.

In this study, we used a life span model to study the subjective perception of creative personality (CP) in emerging, young, middle-aged, and older Hong Kong Chinese adults. We also asked participants to estimate the approximate age by which people develop and lose CP across adulthood. We expected an interesting interplay between internalized age stereotypes and age-related differentiation in beliefs about personality development. Older adults perceived increases in both gains and losses in CP in old age. But they still maintained a similar level of self-perceived CP traits when compared with young participants. Emerging, young, and middle-aged adults were less optimistic about their creativity development into old age. Young adults, in contrast to older adults, believed that gains in CP began and ceased at an earlier age. Positive perceptions of CP in one's aging process may have implications for aging successfully.