Partner and professional support are associated with father-infant bonding: A cross-sectional analysis of mothers, midwives, and child health nurses' influence on primiparous and multiparous fathers of infants in Sweden.

OBJECTIVE: To assess if received professional and social support are associated with father-infant bonding among primiparous (first-time) and multiparous (multi-time) fathers. BACKGROUND: Early father-infant bonding predicts several positive child outcomes. However, while received professional and social support positively impacts fathers' transition into parenthood, little research has tested if these factors are associated with a stronger father-infant bond. METHODS: In total, 499 fathers (296 primiparous and 203 multiparous) of infants (aged 0-12 months) completed a cross-sectional online survey between November 2018 and March 2020. The survey included items related to socio-demographics, having a planned pregnancy, postnatal midwifery support, child health nurse support, child health center attendance, and social support. The parent-infant bonding questionnaire (PBQ) was used to assess the father-infant bond. Multiple linear regression models were estimated for the total sample and based on paternal parity. Missing data were managed through multiple imputation procedures. FINDINGS: Fathers reported fewer bonding disturbances if they received support from their partners, postnatal midwives, child health nurses, and attended more child health visits. Primiparous fathers reported fewer bonding disturbances when receiving support from their partners, postnatal midwives, and the child health nurse. However, multiparous fathers had more bonding disturbances than primiparous fathers and received less professional and partner support. CONCLUSIONS: Receiving more partner and professional support is associated with less father-infant bonding disturbances. To encourage a better father-infant bond, clinicians should invite and support all fathers, regardless of parity, as they transition to parenthood.

Rapid Creation of Knowledge-Balanced Student Groups Using ChatGPT4.

We demonstrated use of ChatGPT4 for efficient group formation in undergraduate medical education. ChatGPT4 created balanced groups considering students' backgrounds in microbiology, physiology, genetics, and immunology considerably faster than manual efforts. Manual refinements included gender balance and discipline distribution. Improvements included ChatGTP's ability to further incorporate GPA and MCAT scores.

Measurement invariance of the Coparenting Relationship Scale (CRS) across 10 countries.

The purpose of this study was to assess the factor structure and the measurement invariance of the Coparenting Relationship Scale (CRS) across 10 countries based on the seven-factor coparenting model (i.e., Coparenting Agreement, Coparenting Closeness, Exposure to Conflict, Coparenting Support, Endorsement of Partner's Parenting; Division of Labor) proposed by Feinberg (2003). The results of research on coparenting from numerous countries have documented its foundational importance for parent mental health, family relationship quality, child development, and psychopathology. Yet, a cross-country perspective is still lacking. Such a perspective can provide insight into which dimensions of coparenting are universally recognized and which are especially prone to variation. A unique multinational data set, comprised of 15 individual studies collected across 10 countries (Belgium, Brazil, China, Israel, Italy, Japan, Portugal, Switzerland, Turkey, USA) in nine languages was established (N = 9,292; 51.1% mothers). Measurement invariance analyses were conducted. A six-factor structure (original seven factors minus Division of Labor) of the measure was consistent across the different contexts and measurement invariance was achieved at the configural level. There was no support for metric or scalar invariance. These findings provide a basis for the CRS to be used across countries and should inspire future quantitative and qualitative research in cross-country coparenting research to understand what aspects are universal and what aspects of coparenting are linked to specific material, relational, or ideational conditions that underlie high-quality coparenting. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Cluster Analysis of Fear of Childbirth, Anxiety, Depression, and Childbirth Self-Efficacy.

OBJECTIVE: To identify clusters of women based on anxiety, depression, fear of birth, and childbirth self-efficacy and factors associated with the clusters. DESIGN: Cross-sectional survey. SETTING: Online in Sweden. PARTICIPANTS: Pregnant women (N = 1,419). METHODS: We collected data through online questionnaires. We included scales to measure anxiety, depression, worries about and fear of birth, and self-efficacy in a kappa-means cluster analysis. We calculated odds ratios with 95% confidence intervals between clusters and background variables. RESULTS: We identified 4 clusters based on severity: Resourceful-Robust, Resourceful-Fearful, Vulnerable-Fearful, and Fragile-Fearful. Participants in the Resourceful-Fearful and Vulnerable-Fearful clusters were more likely to report mental health problems than those in the Resourceful-Robust cluster. Participants in the Vulnerable-Fearful and Fragile-Fearful clusters were more likely to report mental health problems than those in the Resourceful-Robust cluster. Participants in the Fragile-Fearful cluster were more likely to be multiparous, report that their pregnancy was not normal, and prefer cesarean birth than those in the Resourceful-Robust cluster. CONCLUSIONS: Women with childbirth fear may be vulnerable to anxiety and depression during the perinatal period, although the severity might vary. Self-efficacy might be a mediator against mental health problems. Findings demonstrated levels of severity, and the one-size-fits-all approach in Swedish health care may benefit from a more targeted approach for women with fear of childbirth.

Preferences of support and barriers and facilitators to help-seeking in pregnant women with severe fear of childbirth in Sweden: a mixed-method study.

BACKGROUND: There are few support interventions for women with fear of childbirth tailored towards type of fears and parity. To inform the future development of an acceptable and relevant intervention for women with severe fear of childbirth, primary objectives were to examine: (1) pregnant women's experiences of and preferences for support and (2) barriers and facilitators to help-seeking. Secondary objectives were to examine if there are any differences based on pregnant women's parity. METHODS: Pregnant women with a severe fear of childbirth in Sweden completed an online cross-sectional survey between February and September 2022. Severe fear of childbirth was measured using the fear of childbirth scale. Quantitative data were analysed using descriptive and inferential statistics and free answers were analysed using manifest content analysis. A contiguous approach to integration was adopted with qualitative and quantitative findings reported separately. RESULTS: In total, 609 participants, 364 nulliparous and 245 parous women, had severe fear of childbirth. The main category "A twisting road to walk towards receiving support for fear of childbirth" was explored and described by the generic categories: Longing for support, Struggling to ask for support, and Facilitating aspects of seeking support. Over half (63.5%), of pregnant women without planned or ongoing treatment, wanted support for fear of childbirth. Most (60.2%) pregnant women with ongoing or completed fear of childbirth treatment regarded the treatment as less helpful or not at all helpful. If fear of childbirth treatment was not planned, 35.8% of women would have liked to have received treatment. Barriers to help seeking included stigma surrounding fear of childbirth, previous negative experiences with healthcare contacts, fear of not being believed, fear of not being listened to, and discomfort of having to face their fears. Facilitators to help seeking included receiving respectful professional support that was easily available, flexible, and close to home. CONCLUSIONS: Most pregnant women with severe fear of childbirth felt unsupported during pregnancy. Findings emphasise the need to develop individual and easily accessible psychological support for women with severe fear of childbirth, delivered by trained professionals with an empathetic and respectful attitude.

Putting co-creation into practice: lessons learned from developing a midwife-led quality improvement intervention.

BACKGROUND: Integrating evidence-based midwifery practices improves healthcare quality for women and newborns, but an evidence-to-practice gap exists. Co-created quality improvement initiatives led by midwives could bridge this gap, prevent resource waste and ensure intervention relevance. However, how to co-create a midwife-led quality improvement intervention has not been scientifically explored. OBJECTIVE: The objective of this study is to describe the co-creation process and explore the needs and determinants of a midwife-led quality improvement targeting evidence-based midwifery practices. METHODS: A qualitative deductive approach using the Consolidated Framework for Advancing Implementation Science was employed. An analysis matrix based on the framework was developed, and the data were coded according to categories. Data were gathered from interviews, focus group discussions, observations and workshops. New mothers and birth companions (n = 19) were included through convenience sampling. Midwives (n = 26), professional association representatives, educators, policymakers, managers, and doctors (n = 7) were purposely sampled. RESULTS: The co-creation process of the midwife-led Quality Improvement intervention took place in four stages. Firstly, core elements of the intervention were established, featuring a group of midwife champions leading a quality improvement initiative using a train-the-trainers approach. Secondly, the intervention needs, context and determinants were explored, which showed knowledge and skills gaps, a lack of shared goals among staff, and limited resources. However, there was clear relevance, compatibility, and mission alignment for a midwife-led quality improvement at all levels. Thirdly, during co-creation workshops with new mothers and companions, the consensus was to prioritise improved intrapartum support, while workshops with midwives identified enhancing the use of birth positions and perineal protection as key focus areas for the forthcoming Quality Improvement intervention. Lastly, the findings guided intervention strategies, including peer-assisted learning, using existing structures, developing educational material, and building stakeholder relationships. CONCLUSIONS: This study provides a practical example of a co-creation process for a midwife-led quality improvement intervention, which can be relevant in different maternity care settings.

Highlighting Molecular Biology as a Focal Point for Mosquito Research.

ARTICLES DISCUSSED: Chiu, M. Z., Lannon, S., Luchetti, M., Wells, M. B., Andrew, D. J. Dissection and immunostaining of larval salivary glands from Anopheles gambiae mosquitoes. Journal of Visualized Experiments. (175), e62989 (2021). Kojin, B. B., Tsujimoto, H., Jakes, E., O'Leary, S., Adelman, Z. N. Indel detection following CRISPR/Cas9 mutagenesis using high-resolution melt analysis in the mosquito Aedes aegypti. Journal of Visualized Experiments. (175), e63008 (2021). Kumpitak, C., Nguitragool, W., Cui, L., Sattabongkot, J., Bantuchai, S. Detection of Plasmodium sporozoites in Anopheles mosquitoes using an enzyme-linked immunosorbent assay. Journal of Visualized Experiments. (175), e63158 (2021). Taracena, M. et al. Effective oral RNA interference (RNAi) administration to adult Anopheles gambiae mosquitoes. Journal of Visualized Experiments. (181), e63266 (2022). Erlank, E., Venter, N., Koekemoer, L. L. Standard membrane feeding assay for the detection of Plasmodium falciparum infection in Anopheles mosquito vectors. Journal of Visualized Experiments. (183), e63546 (2022).

Bidirectional associations between paternal postpartum depression symptoms and coparenting: A cross-lagged panel model of fathers of infants and toddlers.

BACKGROUND: Finding modifiable predictors of paternal depression symptoms is helpful for developing interventions. The aim is to assess the unidirectional and/or bidirectional associations between paternal postpartum depression symptoms and coparenting among fathers of infants and toddlers. METHODS: Longitudinal data were collected prospectively from 429 fathers of infants aged 0-24 months (median = 8 months) in Sweden, with 6- and 18-month follow-ups. All fathers participated in at least two of three waves of data collection, and multiple imputation was used for missing values. The Edinburgh Postnatal Depression Scale was used to detect depression symptoms (≥10 points), while the Brief Coparenting Relationship Scale measured the coparenting relationship. A cross-lagged panel model was used to estimate the associations between paternal depression symptoms and coparenting relationship quality over time, controlling for several known covariates and COVID-19 exposure. RESULTS: Fathers with higher coparenting scores at Time 1 and 2 had less depression symptoms at Time 3, and fathers with more depression symptoms at Time 2 had lower coparenting scores at Time 3. Plotted probabilities of having at least mild depression symptoms revealed a multifold increase in the probability of depression symptoms at Time 3 for fathers with minimal coparenting scores at Times 1 and 2, respectively, compared to fathers with mean coparenting scores at Times 1 and 2, respectively. LIMITATIONS: Causal links cannot be determined using the current non-experimental study design. Using the EPDS alone may have missed some fathers with depression symptoms. CONCLUSIONS: Clinicians seeking to reduce paternal depression symptoms should help strengthen the coparenting relationship.

Paternal postpartum depression, coparenting, and father-infant bonding: Testing two mediated models using structural equation modeling.

BACKGROUND: Impaired father-infant bonding is a mental health issue that has been understudied. The current study aimed to examine the extent to which fathers' depression symptoms and coparenting relationship, respectively, are associated with infant bonding, as well as how these two variables mediate in the association with infant bonding. METHODS: Cross-sectional data from 612 fathers of infants (0-24 months) were used in this study. Path analyses on two competing mediation models were used to examine the direct and indirect associations between fathers' depression symptoms, coparenting relationship, and infant bonding, after controlling for several known covariates. RESULTS: Whether fathers reported depression symptoms (Model 1) or reported lower levels of coparenting relationships (Model 2), they also reported higher levels of impaired infant bonding, rejection and anger, and anxiety about care after controlling for the covariates. The analysis further found significant indirect associations between father depression symptoms and father-infant bonding via the coparenting relationship (Model 1) as well as significant indirect associations between the coparenting relationship and father-infant bonding via father depression symptoms (Model 2). LIMITATIONS: The cross-sectional data cannot show causal links. Specific efforts were made to recruit fathers with depression symptoms, and therefore prevalence rates may appear skewed. CONCLUSIONS: Decreasing depression symptoms and improving coparenting quality can potentially improve father-infant bonding. Fathers' depression symptoms and coparenting quality can be bi-directionally related in the association with father-infant bonding. Early detection and preventive treatments for paternal depression symptoms and coparenting are warranted.

Midwives and child health nurses' support is associated with positive coparenting for fathers of infants: A cross-sectional analysis.

AIMS AND OBJECTIVES: To examine if the professional support that fathers received from midwives and child health nurses was associated with improvements in fathers' coparenting. A secondary aim was to investigate if there were any support differences between fathers based on parity. BACKGROUND: Stronger coparenting is associated with improved maternal, paternal and child health. It is unclear if routine prenatal and postnatal professional support is associated with improved coparenting in fathers of infants. DESIGN: Cross-sectional online survey. METHODS: In total, 612 fathers of infants (aged 0-24 months) completed baseline data between November 2018 and March 2020. Socio-demographics, pregnancy control variables, social support, professional support, being invited to attend and attending three specific visits for fathers, respectively, and the fathers' coparenting relationship, using the Brief Coparenting Relationship Scale, were assessed. The STROBE checklist was used as the reporting guideline for this study. RESULTS: Fathers' attendance at child health visits, support from the prenatal and postnatal midwife, respectively, and total support from the child health nurse, are associated with more positive coparenting. Primiparous fathers reported more received social and professional support, as well as a more positive coparenting relationship than multiparous fathers. CONCLUSIONS: Receiving clinical support from both midwives and child health nurses is associated with fathers' positive coparenting. All fathers should be invited and encouraged to attend prenatal, postnatal and child health visits to further support their coparenting relationship. Relative to primiparous fathers, multiparous fathers may require targeted and additional clinical support regarding their coparenting relationship. RELEVANCE TO CLINICAL PRACTICE: With fathers becoming more involved in childrearing, having stronger coparenting skills can help them better adapt to their parental roles. Our findings help understand how routine professional support from midwives and child health nurses are experienced among new fathers and that multiparous fathers are in further need of coparenting support.

The Swedish father/non-birthing parent visit: evaluating implementation fidelity among child health nurses one year after voluntary implementation.

Swedish child health centres (CHCs) have created a series of visits for fathers/non-birthing parents. The primary aim was to assess child health nurses' implementation fidelity of the father/non-birthing parent visits, with a secondary aim of exploring predictor variables for fidelity. In 2017, nurses voluntarily implemented a series of father/non-birthing parent visits in Region Stockholm. Nurses (n = 122) completed baseline and 8-12 month follow-up surveys. Multiple imputation was used for missing data. Register data on the number of fathers attending the three-to-five month visit was used. Frequencies of nurses reporting good overall adherence to the home visit, three-to-five week visit, and three-to-five month visit were 86%, 76%, and 68%, respectively. A total of 3,609 fathers attended the three-to-five month visit in 2018, where over half of the visits were at 14 of the 134 CHCs. Multiple linear regression showed that working for a private CHC, seeing more fathers, and nurses' perceptions of receiving enough support predicted higher three-to-five month visit adherence. After nurses saw eight fathers, they were more likely to adhere to the guidelines.

Parenthood and lower risk of suicide in women and men: the total Swedish population followed across adulthood.

BACKGROUND: Previous studies suggest a protective effect of parenthood on suicide, but little is known about how the association may change across the lifespan, or in relation to sex, marital status or occurrence of psychiatric disorders. METHODS: We followed a cohort of over 5 million Swedish women and men, from 1991 to 2011, up to max. age 75, for death by suicide using national registers. Information on childbirths/adoptions, potential confounders and modifying factors were obtained from national registers. We assessed the associations between parenthood and suicide across adulthood using within time-stratified Cox regression models, with parenthood as a time-dependent exposure. RESULTS: Parents had a lower risk of suicide than non-parents across the lifespan, after adjusting for sociodemographic factors. The association was most pronounced in young adults, especially young women, but attenuated with increasing age and converged between sexes in older age groups. The lower risk of suicide over the life course was similar whether parents were married, unmarried or divorced, apart from married men; among them, parents only had a lower risk above age 55. The lower risk in parents was also evident in people with a history of psychiatric hospitalizations, but disappeared from age 55 in this population. CONCLUSION: The lower risk of suicide was present in both parents, was most pronounced in young adulthood and weakened with increasing age. Our results are consistent with a plausible mechanism where feelings of responsibility and connectedness are protective against suicide in parents.

Transgenic Anopheles mosquitoes expressing human PAI-1 impair malaria transmission.

In mammals, the serine protease plasmin degrades extracellular proteins during blood clot removal, tissue remodeling, and cell migration. The zymogen plasminogen is activated into plasmin by two serine proteases: tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA), a process regulated by plasminogen activator inhibitor 1 (PAI-1), a serine protease inhibitor that specifically inhibits tPA and uPA. Plasmodium gametes and sporozoites use tPA and uPA to activate plasminogen and parasite-bound plasmin degrades extracellular matrices, facilitating parasite motility in the mosquito and the mammalian host. Furthermore, inhibition of plasminogen activation by PAI-1 strongly blocks infection in both hosts. To block parasite utilization of plasmin, we engineered Anopheles stephensi transgenic mosquitoes constitutively secreting human PAI-1 (huPAI-1) in the midgut lumen, in the saliva, or both. Mosquitoes expressing huPAI-1 strongly reduced rodent and human Plasmodium parasite transmission to mosquitoes, showing that co-opting plasmin for mosquito infection is a conserved mechanism among Plasmodium species. huPAI-1 expression in saliva induced salivary gland deformation which affects sporozoite invasion and P. berghei transmission to mice, resulting in significant levels of protection from malaria. Targeting the interaction of malaria parasites with the fibrinolytic system using genetically engineered mosquitoes could be developed as an intervention to control malaria transmission.

A qualitative study of Swedish fathers' experiences of becoming a father during the COVID-19 pandemic.

INTRODUCTION: Expectant fathers want to participate in perinatal care. COVID-19 policies restrict their access to care, but it is unknown how these policies have affected them. The aim of this study is to explore the perinatal care given to and wanted by expectant and new fathers during the COVID-19 pandemic in Sweden. METHODS: The current study used an inductive qualitative design where 14 expectant or new fathers participated in a video- or telephone-based semi-structured interview. Interviews lasted 20 minutes, on average. The collected data were analyzed using content analysis. RESULTS: Two main themes were reported: 1) 'Being left out, but trying to remain positive', and 2) immediate consequences related to restrictions. Expectant fathers were not able to attend as many perinatal visits as they wanted to, due to the COVID-19 restrictions on non-birthing parents. Expectant fathers regretted and felt discouraged that they could not support their partner during visits and not follow their baby's growth and development. Furthermore, they faced uncertainties and stress regarding whether or not they could attend the birth of their child. Fathers reported how their exclusion negatively impacted the entire family. CONCLUSIONS: Expectant and new fathers felt that their level of participation in prenatal care was negatively impacted by the Swedish policies imposed on them during the COVID-19 pandemic. Fathers were physically and emotionally excluded, resulting in receiving little direct care support, and lacked companionship with other parents. Fathers provided suggestions and alternatives on how to increase their participation with individual midwives and from an organizational level.

Ribbon boosts ribosomal protein gene expression to coordinate organ form and function.

Cell growth is well defined for late (postembryonic) stages of development, but evidence for early (embryonic) cell growth during postmitotic morphogenesis is limited. Here, we report early cell growth as a key characteristic of tubulogenesis in the Drosophila embryonic salivary gland (SG) and trachea. A BTB/POZ domain nuclear factor, Ribbon (Rib), mediates this early cell growth. Rib binds the transcription start site of nearly every SG-expressed ribosomal protein gene (RPG) and is required for full expression of all RPGs tested. Rib binding to RPG promoters in vitro is weak and not sequence specific, suggesting that specificity is achieved through cofactor interactions. Accordingly, we demonstrate Rib's ability to physically interact with each of the three known regulators of RPG transcription. Surprisingly, Rib-dependent early cell growth in another tubular organ, the embryonic trachea, is not mediated by direct RPG transcription. These findings support a model of early cell growth customized by transcriptional regulatory networks to coordinate organ form and function.

Family life starts at home: Fathers' experiences of a newly implemented Swedish home-based postnatal care model - an interview study.

OBJECTIVE: To explore and describe fathers' experiences of a newly implemented Swedish home-based postnatal care model. DESIGN: A descriptive cross-sectional qualitative study was conducted as a part of a larger study. SETTING: Families who qualified to be discharged early were offered to participate in a postnatal home-based model of midwifery care by a hospital in Stockholm, Sweden. PARTICIPANTS AND MEASUREMENTS: In total, 16 fathers participated in a semi-structured telephone interview, averaging 43 min. Data were analyzed using systematic text condensation. FINDINGS: Three major themes emerged: To decide on home- or hospital-based postnatal care - a matter of safety, To be offered professional midwifery postnatal support at home, and To be at home helped fathers to navigate parenthood. Fathers appreciated the home-based postnatal care and felt safe because of the received professional support from midwives. KEY CONCLUSIONS: Home-based postnatal care was valued by fathers whose partner had a non-complicated vaginal birth because they felt safe in their home environment and supported by midwives. The home environment aided fathers in supporting their partners and developing a father-infant bond. IMPLICATIONS FOR PRACTICE: Home-based postnatal care was valued by fathers and should be considered an option for new families. To offer home-based postnatal care may result in less overcrowded postnatal wards. Midwives need to enable fathers' participation and support their parental role regardless of where the care takes place.

Facilitators and barriers when implementing a new clinical visit for fathers at Stockholm County child health centres: A qualitative study of nurse-managers at top-performing centres.

BACKGROUND: In 2017, a new clinical visit for non-birthing parents (e.g. fathers) of three-to-five month old infants was implemented in Stockholm County. In 2018, 19 out of 134 child health centres (CHCs) saw 62% of all fathers at the father visit and these CHCs had the highest fidelity scores. The aim of the current study was to assess nurse-managers' perceptions of facilitators and barriers to implementating the father visit. METHODOLOGY: Semi-structured interviews were conducted with nurse-managers from seven CHCs. Interviews were audio-recorded, transcribed and then analysed using systematic text condensation. Hasson's modified version of the Conceptual Framework for Implementation Fidelity was used to interpret the findings. RESULTS: Six themes emerged during the analysis, which were sorted into perceived facilitators and barriers regarding the implementation of the father visit. Facilitators included: 1) nurses are facilitators of change, 2) fathers are worthy of change, 3) a child has two parents and 4) effective cooperation. The barriers included: 5) money talks and 6) nurses under pressure. CONCLUSION: These findings provide insight into factors that moderated the implementation fidelity of the father visits at CHCs with some of the highest levels of father attendance. This provides a model of good practice that can be applied to other settings to maximise fidelity, as well as increase attendance at father visits. Findings also provide insight into the relationship between potential moderators and implementation fidelity.

A dreaded delight: A longitudinal qualitative interview study of paternal childbirth fear during the transition to fatherhood.

INTRODUCTION: Childbirth is a life-changing event where fathers desire being involved. As fathers take a more active role, fear of childbirth can arise. The aim of this study was to explore fathers' experiences of childbirth fear during pregnancy and after the birth of their baby. METHODS: This was a qualitative longitudinal prospective study that included 14 interviews with seven fathers, one during the prenatal period, and one follow-up interview after childbirth. Data were analyzed using thematic analysis according to Braun and Clark. RESULTS: The main theme 'Being vulnerable during the transition to fatherhood' was based on the perception of childbirth as risky with threats toward the woman's and baby's health, not being able to give and receive enough support, unable to handle the unknown process of birth, and not being a good father. Helpful strategies for coping with fear were to talk about fear, to learn more about childbirth and techniques on how to handle fear, and to avoid dealing with fear. Fathers' fear of childbirth changed after the birth of their baby. Their thoughts of another childbirth did not evoke the same strong feelings of fear. Issues important for the reduction of childbirth fear were: receiving professional support, actively taking part in the childbirth process, and the partner having an uncomplicated birth. CONCLUSIONS: Fathers with childbirth fear regarded childbirth as risky, but they expressed helpful coping strategies. After the birth of their baby, they became less fearful. Quality of fear-reducing support to expectant fathers may influence how they cope with their transition into fatherhood.

Dissection and Immunostaining of Larval Salivary Glands from Anopheles gambiae Mosquitoes.

Mosquito salivary glands (SGs) are a requisite gateway organ for the transmission of insect-borne pathogens. Disease-causing agents, including viruses and the Plasmodium parasites that cause malaria, accumulate in the secretory cavities of SG cells. Here, they are poised for transmission to their vertebrate hosts during a subsequent blood meal. As adult glands form as an elaboration of larval SG duct bud remnants that persist beyond early pupal SG histolysis, the larval SG is an ideal target for interventions that limit disease transmission. Understanding larval SG development can help develop a better understanding of its morphology and functional adaptations and aid in the assessment of new interventions that target this organ. This video protocol demonstrates an efficient technique for isolating, fixing, and staining larval SGs from Anopheles gambiae mosquitoes. Glands dissected from larvae in a 25% ethanol solution are fixed in a methanol-glacial acetic acid mixture, followed by a cold acetone wash. After a few rinses in phosphate-buffered saline (PBS), SGs can be stained with a broad array of marker dyes and/or antisera against SG-expressed proteins. This method for larval SG isolation could also be used to collect tissue for in situ hybridization analysis, other transcriptomic applications, and proteomic studies.

Secrets of secretion-How studies of the Drosophila salivary gland have informed our understanding of the cellular networks underlying secretory organ form and function.

Secretory organs are critical for organismal survival. Yet, the transcriptional regulatory mechanisms governing their development and maintenance remain unclear for most model secretory organs. The Drosophila embryonic salivary gland (SG) remedies this deficiency as one of the few organs wherein direct connections from the expression of the early patterning genes to cell specification to organ architecture and functional specialization can be made. Few other models of secretion can be accorded this distinction. Studies from the past three decades have made enormous strides in parsing out the roles of distinct transcription factors (TFs) that direct major steps in furnishing this secretory organ. In the first step of specifying the salivary gland, the activity of the Hox factors Sex combs reduced, Extradenticle, and Homothorax activate expression of fork head (fkh), sage, and CrebA, which code for the major suite of TFs that carry forward the task of organ building and maintenance. Then, in the second key step of building the SG, the program for cell fate maintenance and morphogenesis is deployed. Fkh maintains the secretory cell fate by regulating its own expression and that of sage and CrebA. Fkh and Sage maintain secretory cell viability by actively blocking apoptotic cell death. Fkh, along with two other TFs, Hkb and Rib, also coordinates organ morphogenesis, transforming two plates of precursor cells on the embryo surface into elongated internalized epithelial tubes. Acquisition of functional specialization, the third key step, is mediated by CrebA and Fkh working in concert with Sage and yet another TF, Sens. CrebA directly upregulates expression of all of the components of the secretory machinery as well as other genes (e.g., Xbp1) necessary for managing the physiological stress that inexorably accompanies high secretory load. Secretory cargo specificity is controlled by Sage and Sens in collaboration with Fkh. Investigations have also uncovered roles for various signaling pathways, e.g., Dpp signaling, EGF signaling, GPCR signaling, and cytoskeletal signaling, and their interactions within the gene regulatory networks that specify, build, and specialize the SG. Collectively, studies of the SG have expanded our knowledge of secretory dynamics, cell polarity, and cytoskeletal mechanics in the context of organ development and function. Notably, the embryonic SG has made the singular contribution as a model system that revealed the core function of CrebA in scaling up secretory capacity, thus, serving as the pioneer system in which the conserved roles of the mammalian Creb3/3L-family orthologues were first discovered.