Exploring the gap between needs and practice in facilitating breastfeeding within the neonatal intensive care setting : An Italian survey on organizational factors

Introduction: The system-level factors of the neonatal intensive care unit work environment contribute to breastfeeding promotion in the preterm population. The aim of this study was to investigate the operative policies related to breastfeeding support in a sample of Italian Neonatal Intensive Care Units. Materials and Methods: A multicenter cross-sectional survey was conducted, including a sample of 17 head nurses. The items of the questionnaire investigated the following areas: breastfeeding policies, staff education, family centered care, and breastfeeding promotion and support both in the neonatal intensive care units and after discharge. Results: Written breastfeeding policies were available for staff in all the neonatal intensive care units, most commonly addressing procedures related to skin-to-skin contact, human milk expression, and preterm infant breastfeeding. Most of the neonatal intensive care units correctly advised the mothers to initiate milk expression within 6 h from delivery and to pump milk at least 6 times/days. Breastfeeding training for the nursing staff was planned in the majority of the neonatal intensive care units although according to different schedules. With regard to the family centered care implementation, time restrictions were present in seven neonatal intensive care units, mostly occurring during the night shift, and the morning hours concomitantly with medical rounds. Moreover, in the majority of the investigated neonatal intensive care units, the parents were asked to leave the ward when their infant underwent a major invasive procedure or during the nurse/physician shift change report. With regard to breastfeeding promotion and support, eight neonatal care units had a multidisciplinary team with several health care professionals and 10 provided information about community-based support services. Most of the units assessed breastfeeding after discharge. Conclusion: Based on the present findings, enrolled Neonatal Intensive Care Units appear to provide breastfeeding-supportive environments with special regard to breastfeeding policies, milk expression practices, interprofessional collaboration, and continuity of care. Health care professionals should exert efforts to ensure continuous and updated breastfeeding staff education and promote parent-infant closeness and family centered care

Human exonuclease 1 role in response to UV irradiation

DNA damage checkpoints are surveillance mechanisms that monitor the integrity of the genome. Nucleotide excision repair (NER) is a DNA repair mechanism that cells use to remove UV-induced DNA lesions. Previous publication from our laboratory demonstrated that recognition and processing of UV-induced damage by NER is required for proper activation of checkpoint through interactions between NER proteins and checkpoint factors in yeast and human primary fibroblasts. From a two hybrid screening in yeast exonuclease 1 (Exo1) was identified as a 9-1-1 complex interactor. Exo1 is a 5\u2019-3\u2019 exonuclease and 5'-flap-endonuclease with many different roles in DNA metabolism such as meiotic and mitotic recombination, mismatch repair and telomere processing. Characterization of an exo1 yeast deleted strain has shown that this protein is involved in the early steps of UV-induced DNA damage checkpoint. In human cells EXO1 is present as two isoforms named hEXO1a and hEXO1b genetarated by alternative splicing. We are analyzing the role of EXO1 in checkpoint activation in response to UV-C damage in human cells: using siRNA against both a and b isoform of hEXO1 in G1 cells we were able to observe a defect in Chk1 and p53 phosphorylation induced by UV-C irradiation

Haspin regulates Ras localization to promote Cdc24-driven mitotic depolarization

Cell polarization is of paramount importance for proliferation, differentiation, development, and it is altered during carcinogenesis. Polarization is a reversible process controlled by positive and negative feedback loops. How polarized factors are redistributed is not fully understood and is the focus of this work. In Saccharomyces cerevisiae, mutants defective in haspin kinase exhibit stably polarized landmarks and are sensitive to mitotic delays. Here, we report a new critical role for haspin in polarisome dispersion; failure to redistribute polarity factors, in turn, leads to nuclear segregation defects and cell lethality. We identified a mitotic role for GTP-Ras in regulating the local activation of the Cdc42 GTPase, resulting in its dispersal from the bud tip to a homogeneous distribution over the plasma membrane. GTP-Ras2 physically interacts with Cdc24 regulateing its mitotic distribution. Haspin is shown to promote a mitotic shift from a bud tip-favored to a homogenous PM fusion of Ras-containing vesicles. In absence of haspin, active Ras is not redistributed from the bud tip; Cdc24 remains hyperpolarized promoting the activity of Cdc42 at the bud tip, and the polarisome fails to disperse leading to erroneously positioned mitotic spindle, defective nuclear segregation, and cell death after mitotic delays. These findings describe new functions for key factors that modulate cell polarization and mitotic events, critical processes involved in development and tumorigenesis

TLS Polymerases are involved in processing of EXO1-dependent lesions after UV-induced damage

UV light mainly damages DNA by generating CPDs and 6-4PP photoproducts, which are responsible for the pathological effects of sunlight. In a healthy organism, such DNA helix distorting lesions are removed by Nucleotide Excision Repair (NER), a multistep process. Mutations in NER genes cause the onset of severe pathologies. The principal symptom common to all diseases is the strong sensitivity to UV. A high predisposition to tumors development arises in xeroderma pigmentosum (XP) patients, while neurological dysfunctions have been observed in both XP and Cockayne syndrome patients. Upon DNA damage sensing, checkpoints are activated allowing a block or delay of cell cycle progression to ensure repair of the DNA lesions. Intriguingly, while in normal cells UV irradiation activates DNA damage checkpoints in all phases of the cell cycle NER yeast mutant strains and human fibroblasts derived from XP patients fail activate the checkpoint in G1 and G2. Recently, we demonstrated that the checkpoint response to UV light in cells that are not actively replicating their genome requires prior processing of the UV lesions. This involves NER factors but also the Exo1 nuclease. In particular, acting on NER intermediates, Exo1 generates structures containing long tracts of ssDNA in response to UV irradiation. This role of Exo1 is only observed at a subset of problematic lesions that cannot properly repaired by canonic NER. It is these Exo1-induced structures that provide the signal for checkpoint activation both in yeast and human non-replicating cells. The essential role of Exo1 in UV-induced checkpoint activation in vivo has been recently supported by in vitro reconstitution of the activation pathway. What are the problematic lesions that require EXO1 activity is still unknown. We hypothesized that Closely Opposing UV Lesions (COLs) on the two DNA strands could exist and may be a likely candidate. This scenario would require TLS polymerases bypass during repair synthesis step. Therefore, we are investigating Y-family polymerase recruitment at EXO1-positive local UV damage sites (LUDs). We found that Pol h is recruited at both EXO1-positive and EXO1-negative LUDs, while Pol \u3b9 and\uf020Pol \u3ba always co-localize with the nuclease\uf02e Using the CRISPR-Cas9 system, we generated EXO1 knock out cell lines that demonstrated a requirement for EXO1 in Pol \u3b9 and\uf020Pol \u3ba recruitment, consistently with our working model\uf02e Finally, when we silenced TLS polymerases we observed a hyper-activation of UV-induced DNA damage checkpoint, suggesting that EXO1 continues to process UV damaged DNA enlarging the gap and eventually producing DSBs. TLS polymerases, thus are crucial to prevent dangerous situations in non-replicating UV irradiated cells

The B subunit of the DNA polymerase alpha-primase complex in Saccharomyces cerevisiae executes an essential function at the initial stage of DNA replication

The four-subunit DNA polymerase \uce\ub1-primase complex is unique in its ability to synthesize DNA chains de novo, and some in vitro data suggest its involvement in initiation and elongation of chromosomal DNA replication, although direct in vivo evidence for a role in the initiation reaction is still lacking. The function of the B subunit of the complex is unknown, but the Saccharomyces cerevisiae POL12 gene, which encodes this protein, is essential for cell viability. We have produced different pol12 alleles by in vitro mutagenesis of the cloned gene. The in vivo analysis of our 18 pol12 alleles indicates that the conserved carboxy-terminal two-thirds of the protein contains regions that are essential for cell viability, while the more divergent NH2-terminal portion is partially dispensable. The characterization of the temperature-sensitive pol12-T9 mutant allele demonstrates that the B subunit is required for in vivo DNA synthesis and correct progression through S phase. Moreover, reciprocal shift experiments indicate that the POL12 gene product plays an essential role at the early stage of chromosomal DNA replication, before the hydroxyurea-sensitive step. A model for the role of the B subunit in initiation of DNA replication at an origin is presented

Breastfeeding difficulties and risk for early breastfeeding cessation

Although breast milk is the normative feeding for infants, breastfeeding rates are lower than recommended. We investigated breastfeeding difficulties experienced by mothers in the first months after delivery and their association with early breastfeeding discontinuation. We conducted a prospective observational study. Mothers breastfeeding singleton healthy term newborns at hospital discharge were enrolled and, at three months post-delivery, were administered a questionnaire on their breastfeeding experience. Association among neonatal/maternal characteristics, breastfeeding difficulties and support after hospital discharge, and type of feeding at three months was assessed using multivariate binary logistic regression analysis. We enrolled 792 mothers, 552 completed the study. Around 70.3% of mothers experienced breastfeeding difficulties, reporting cracked nipples, perception of insufficient amount of milk, pain, and fatigue. Difficulties occurred mostly within the first month. Half of mothers with breastfeeding issues felt wellsupported by health professionals. Maternal perception of not having a sufficient amount of milk, infant\u2019s failure to thrive, mastitis, and the return to work were associated with a higher risk of nonexclusive breastfeeding at three months whereas vaginal delivery and breastfeeding support after hospital discharge were associated with a decreased risk. These results underline the importance of continued, tailored professional breastfeeding support

Maternal views on facilitators of and barriers to breastfeeding preterm infants

Background: The supply of breast milk to preterm infants tends to occur at a lower rate than that recorded among term infants. We aimed to investigate the facilitators of and barriers to breastfeeding during hospital stay according to the experiences of mothers that gave birth to premature infants requiring admission to neonatal intensive care unit. Methods: A cross-sectional questionnaire survey was conducted. Mothers who had delivered a newborn with a gestational age 6433 weeks requiring intensive care, entered the study. Basic subjects' characteristics and infant feeding practices were also recorded. Results: A total of 64 mothers were enrolled, leading to a total of 81 infants. At discharge, any breastfeeding was recorded in 66% of infants, with 27% of those infants being exclusively breastfed. Any infant was exclusively fed directly at the breast. Most mothers experienced adequate support during their infant's hospitalization and reported satisfaction with breastfeeding. Almost all mothers felt that feeding their infant human milk was beneficial for the infant's health. Thirty percent of the mothers reported that they had experienced some obstacles to breastfeeding. Specifically, infants born to mothers who experienced difficulties in pumping breast milk (OR = 4.6; CI 1.5-13.9) or in providing an adequate amount of milk to the infant (OR = 3.57; CI 1.1-11.5) were at higher risk of being fed with formula at discharge. Conclusions: On the basis of the present results, health care professionals should target their efforts to optimize breastfeeding support for mothers of premature infants admitted to level III care, especially by improving breast milk production and endorsing direct breastfeeding

Exploring the Emotional Breastfeeding Experience of First-Time Mothers : Implications for Healthcare Support

Background: Among breastfeeding determinants, the unique emotional breastfeeding experience has been poorly explored. The present study aimed to investigate the emotional breastfeeding experience in a cohort of first-time mothers. Materials and methods: We conducted a prospective observational study that enrolled primiparas having delivered singleton healthy term newborns, and exclusively breastfeeding at hospital discharge. At 3 months post-delivery mothers accessed an online questionnaire investigating their emotional breastfeeding experience. The chi-squared test was used to assess the association between the feelings experienced during breastfeeding and feeding outcomes at 3 months. Results: Out of the 421 enrolled mothers, 273 (65%) completed the questionnaire. At 3 months post-delivery exclusive breastfeeding was reported by a 66% of mothers, a 19% reported complementary feeding, and a 15% of mothers reported exclusive formula feeding. Breastfeeding experience was described as positive by 62% of mothers although breastfeeding difficulties were reported by 80% of the mothers. The mothers that had experienced fear, sadness, anger or concern during breastfeeding showed a significant higher exclusive formula feeding rate at 3 months post-delivery than those who did not (25.5 vs. 12.8%, p = 0.021; 28.6 vs. 13.4%, p = 0.02; 40 vs. 13.4%, p = 0.005; 20.5 vs. 11.8%, p = 0.049, respectively). An 85% of mothers stated that their breastfeeding experience was different from what they would have expected, blaming for this discrepancy the occurrence of difficulties during breastfeeding and the complexity of breastfeeding itself (50%), pain experience (8%), being dependent from the baby (6%), and breastfeeding failure (11%). A total of 25% of mothers, however, reported they found breastfeeding to be a much more positive experience than what they had expected. Conclusion: Breastfeeding care should include a tailored emotional support of first time-mothers in addition to the implementation of their breastfeeding knowledge and skills