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Introduction Despite support for parenting being already recognised as a priority, there remains a paucity of evidence on how to facilitate its adoption in regular visits of maternal and child health primary care (PC). We describe the protocol for a study to assess the effect of an innovative universal Touchpoints-based intervention—‘Crescer em Grande!’ (CeG!) - when supporting the process of transition to parenthood and early infancy, at multiple PC units. Methods and analysis A cluster-randomised trial will be conducted in 12 PC units (clusters) from the Lisbon metropolitan area, Portugal. Participants will be a minimum of three family physicians and one nurse/unit, as well as 216 expecting parents and future babies until 18 months who are using the PC services. Sites will be randomised to either the CeG! or usual care. The CeG! will consist of: (1) the integration of the Touchpoints approach in PC maternal and well-child visits, with the support of 28 leaflets for parents to file in a folder; plus (2) training for PC providers on how to perform the CeG! into existing practice. Parents will be required to fill in questionnaires at point throughout their child’s 18-month, mostly online. The primary outcome will be the self-perception of parental competence (Parenting Sense of Competence Scale). Other outcomes include: family functioning, couple dynamics, mental health, well-being/quality of life, psychological experience of pregnancy, attachment, child development. Acceptability, satisfaction and feasibility of CeG! will also be obtained from providers’ and parents’ perspectives. Costs associated with delivering the CeG! will be calculated. Study analyses will be under the principle of intention-to-treat. Ethics and dissemination Approval was obtained from the Ethics Committee of the Regional Health Administration. The results will be shared with participants and disseminated via peer-reviewed published papers, presentations at scientific and professional conferences. Trial registration number NCT90692907

In the present study, we standardized processes of cloning and purification of recombinant bovine interleukin-8 (rbIL-8) from bacterial culture and assessed its biological activity in Holstein cattle. Plasmid containing a subclone of bovine IL-8 was expressed using Escherichia coli BL21 and cell lysate was purified by chromatography. The presence of rbIL-8 was assessed by Western blot analyses and function was confirmed in vitro using a chemotaxis chamber. Based on optical density values, chemoattractant properties of rbIL-8 were 10-fold greater compared with control wells. Two in vivo studies were conducted to assess the biological activity of rbIL8. For study 1, one-year-old Holstein heifers (n = 20) were randomly allocated to receive a single intravaginal administration containing 1,125 µg of rbIL-8 diluted in 20 mL of saline solution (rbIL-8, n = 10) or a single intravaginal administration of 20 mL of saline solution (control, n = 10). For study 2, nonpregnant lactating Holstein cows (n = 31) were randomly allocated to receive an intrauterine administration with 1,125 µg of rbIL-8 diluted in 20 mL of saline solution (rbIL-8, n = 11), a positive control consisting of resin-purified lysate of E. coli BL21 not transfected with the plasmid coding for rbIL-8 diluted in 20 mL of saline solution (E. coli, n = 10), and a negative control administered with 20 mL of saline solution (control, n = 10). An increase in vaginal neutrophils was observed in heifers treated with rbIL-8 within 3 h of treatment, but not in control heifers. Additionally, intrauterine administration of rbIL-8 increased the proportion of PMN cells in uterine cytological samples from 3.5% before treatment to 75.8% 24 h later-an increase that was not observed in the negative control group and cows treated with resin-purified lysate of E. coli. To further evaluate the effect of local and systemic rbIL-8 stimulation on the dynamics of circulating white blood cells, a third study was conducted. In study 3, nonpregnant 8-mo-old Holstein heifers (n = 30) were randomly allocated into 1 of 3 treatment groups: intravenous rbIL-8 (1,125 µg of rbIL-8 diluted in 5 mL of saline solution, n = 10); intravaginal rbIL-8 (1,125 µg of rbIL-8 diluted in 20 mL of saline solution; n = 10); or intravaginal saline (20 mL of saline solution, n = 10). Intravenous injection of rbIL-8 resulted in a transient increase in rectal temperature, which was greater at 2 h after treatment compared with cows treated intravaginally with rbIL-8 or heifers treated with saline solution. Heifers treated with rbIL-8 intravenously displayed a marked reduction in neutrophils, basophils, lymphocytes, and monocytes within the first 4 h posttreatment compared with heifers treated intravaginally. However, at 6 h after treatment, heifers treated with rbIL-8 intravenously displayed a rebound in white blood cell counts caused by an increase in neutrophil counts. These results show that the presented purification method is effective and results in biologically active rbIL-8 that can be used safely to m...

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