Fluorescently Tagged Verticillium dahliae to Understand the Infection Process on Cotton (Gossypium hirsutum) and Weed Plant Species.

Verticillium wilt is a soil-borne disease caused by distinct vegetative compatibility groups (VCG) of the fungus Verticillium dahliae. Defoliating (VCG 1A) and non-defoliating (VCG 2A) pathotypes of V. dahliae have contributed to yield losses of cotton production in Australia. To study the virulence and the infection process of V. dahliae on cotton, two isolates, one representing each VCG, have been transformed with fluorescent protein genes. The transformants maintained their ability to infect the host, and both strains were observed to move through the plant vasculature to induce wilt symptoms. Furthermore, virulence testing suggests that the cotton V. dahliae strains can endophytically colonise common weed plant species found in the Australian landscape, and that is contrasted by their ability to infect and colonise native tobacco plants. The fluorescently labelled strains of V. dahliae not only allowed us to gain a thorough understanding of the infection process but also provided a method to rapidly identify recovered isolates from host colonisation studies.

Surveillance, Diversity and Vegetative Compatibility Groups of Fusarium oxysporum f. sp. vasinfectum Collected in Cotton Fields in Australia (2017 to 2022)

Cotton (Gossypium hirsutum) is a billion-dollar crop in regional New South Wales (NSW) and Queensland, Australia. Fusarium wilt (FW) caused by Fusarium oxysporum f. sp. vasinfectum (Fov) is an economically important disease. Initial disease losses of up to 90% when the disease was first detected resulted in fields being taken out of cotton production. The disease is now well-managed due to the adoption of highly resistant varieties. However, annual disease surveys recently revealed that the disease dynamic has changed in the past few seasons. With relatively mild and wet weather conditions during the 2021/22 growing season, FW was detected in eight surveyed valleys in NSW and Queensland, with the disease incidence as high as 44.5% and 98.5% in individual fields in early and late seasons, respectively. Fov is genetically distinct and evolved from local Fusarium oxysporum strains. Additionally, the pathogen was reported to evolve rapidly under continuous cotton cropping pressure. However, our knowledge of the genetic composition of the prevailing population is limited. Sequences of the translation elongation factor alpha 1 (TEF1) revealed that 94% of Fusarium isolates recovered from FW-infected cotton were clustered together with known Australian Fov and relatively distant related to overseas Fov races. All these isolates, except for nine, were further confirmed positive with a specific marker based on the Secreted in Xylem 6 (SIX6) effector gene. Vegetative compatibility group (VCG) analyses of 166 arbitrarily selected isolates revealed a predominance of VCG01111. There was only one detection of VCG01112 in the Border Rivers valley where it was first described. In this study, the exotic Californian Fov race 4 strain was not detected using a specific marker based on the unique Tfo1 insertion in the phosphate (PHO) gene. This study indicated that the prevalence and abundance of Fov across NSW and Queensland in the past five seasons was probably independent of its genetic diversity.

The IBLCE exam: candidate experience, motivation, study strategies used and predictors of success.

Background: Optimising breastfeeding rates is a public health priority. Studies have shown that all forms of extra breastfeeding support increase breastfeeding rates, including support provided by trained health professionals. International Board Certified Lactation Consultants (IBCLCs) are trained healthcare professionals in the clinical management of breastfeeding and human lactation. The IBCLC certification is a sought-after credential and can only be obtained after passing the exam administered by the International Board of Lactation Consultant Examiners (IBLCE). In Slovenia and Croatia, the IBLCE exam has been offered since 2006 and 2009, respectively. In this study, our aim was to 1) determine which candidate characteristics are associated with a passing grade on the IBLCE exam; and 2) analyse differences between candidates from Slovenia and Croatia, given Slovenians' higher achievements in the past. Methods: In February, 2017, a 4-page, 36-question survey was sent via Survey Monkey to the available email addresses of all past IBLCE exam candidates in Croatia and Slovenia. Questions covered sociodemographic data, breastfeeding education, exam preparation, motivation and experience taking the IBLCE exam. Results: Ninety-two participants completed the online survey: 36 from Croatia and 55 from Slovenia, giving a response of 47 and 52%, respectively. No significant difference was found in pass rates between the two countries, despite Slovenians being younger and spending more time observing normal breastfeeding dyads. Variables found to be significantly more common among respondents who passed the IBLCE exam included: attending breastfeeding conferences/symposiums, using a breastfeeding atlas and studying with others. Statistical predictors of IBLCE exam success were: number of hours of bedside teaching, perceived clarity of photographs and breastfeeding conference/symposium attendance. Respondents who reported that they had attended a breastfeeding conference/symposium, had less hours of bedside teaching and perceived exam photographs as completely clear, were 7.49 (95% CI 2.26, 24.84), 0.48 (95% CI 0.28, 0.82), and 3.49 (95% CI 1.17, 10.41) times more likely to pass the exam, respectively. Conclusion: Breastfeeding conference attendance, less bedside teaching and perceived clarity of exam photographs may be predictors of IBLCE exam success. Further studies on larger samples of exam candidates are required to confirm our findings and determine other factors associated with passing the IBLCE exam.

Speaking out on safe sleep: evidence-based infant sleep recommendations.

The American Academy of Pediatrics (AAP) issued recommendations in 2005 and 2011 to reduce sleep-related infant death, which advise against all bedsharing for sleep. These recommendations overemphasize the risks of bedsharing, and this overemphasis has serious unintended consequences. It may result in increased deaths on sofas as tired parents try to avoid feeding their infants in bed. Current evidence shows that other risks are far more potent, such as smoking, shared sleep on sofas, sleeping next to impaired caregivers, and formula feeding. The emphasis on separate sleep is diverting resources away from addressing these critical risk factors. Recommendations to avoid bedsharing may also interfere with breastfeeding. We examine both the evidence behind the AAP recommendations and the evidence omitted from those recommendations. We conclude that the only evidence-based universal advice to date is that sofas are hazardous places for adults to sleep with infants; that exposure to smoke, both prenatal and postnatal, increases the risk of death; and that sleeping next to an impaired caregiver increases the risk of death. No sleep environment is completely safe. Public health efforts must address the reality that tired parents must feed their infants at night somewhere and that sofas are highly risky places for parents to fall asleep with their infants, especially if parents are smokers or under the influence of alcohol or drugs. All messaging must be crafted and reevaluated to avoid unintended negative consequences, including impact on breastfeeding rates, or falling asleep in more dangerous situations than parental beds. We must realign our resources to focus on the greater risk factors, and that may include greater investment in smoking cessation and doing away with aggressive formula marketing. This includes eliminating conflicts of interest between formula marketing companies and organizations dedicated to the health of children.

Colostrum avoidance, prelacteal feeding and late breast-feeding initiation in rural Northern Ethiopia.

OBJECTIVE: To identify specific cultural and behavioural factors that might be influenced to increase colostrum feeding in a rural village in Northern Ethiopia to improve infant health. DESIGN: Background interviews were conducted with six community health workers and two traditional birth attendants. A semi-structured tape-recorded interview was conducted with twenty mothers, most with children under the age of 5 years. Variables were: parental age and education; mother's ethnicity; number of live births and children's age; breast-feeding from birth through to weaning; availability and use of formula; and descriptions of colostrum v. other stages of breast milk. Participant interviews were conducted in Amharic and translated into English. SETTING: Kossoye, a rural Amhara village with high prevalence rates of stunting: inappropriate neonatal feeding is thought to be a factor. SUBJECTS: Women (20-60 years of age) reporting at least one live birth (range: 1-8, mean: ∼4). RESULTS: Colostrum (inger) and breast milk (yetut wotet) were seen as different substances. Colostrum was said to cause abdominal problems, but discarding a portion was sufficient to mitigate this effect. Almost all (nineteen of twenty) women breast-fed and twelve (63 %) reported ritual prelacteal feeding. A majority (fifteen of nineteen, 79 %) reported discarding colostrum and breast-feeding within 24 h of birth. Prelacteal feeding emerged as an additional factor to be targeted through educational intervention. CONCLUSIONS: To maximize neonatal health and growth, we recommend culturally tailored education delivered by community health advocates and traditional health practitioners that promotes immediate colostrum feeding and discourages prelacteal feeding.

How ethical is ethical research? Recruiting marginalized, vulnerable groups into health services research.

AIM: This paper is a discussion of how tensions in research ethics guidelines can have an impact on the recruitment of marginalized, vulnerable groups of people into health services research. Background. Research ethics guidelines in the United Kingdom (UK) predominantly reflect requirements for clinical trials and medical research but apply to all types of research conducted in the National Health Service. Guidelines emphasize the process of informed consent to protect individuals' autonomy and rights, and avoid harm to vulnerable people. This focus, however, can result in the exclusion of and discrimination against marginalized groups at a time when user involvement in research and health service developments is increasingly emphasized in policy. METHOD: This paper is based on a critical reflection on tensions observed in current research ethics guidelines in the UK. These tensions became apparent following recruitment of nine postnatal drug-using women to a qualitative phase of an evaluation in 2005 of a specialist health visitor service. FINDINGS: The main ethical tensions identified concerned the sampling and recruitment of marginalized groups, which can affect the validity and reliability of research findings. Consideration needs to be given to sampling and accessing such groups and assessing the capacity of individuals to give informed consent. This is to avoid coercion and exploitation of vulnerable individuals and groups in research, increase validity and reliability, and avoid pre-emptive exclusion of such groups in the research design. CONCLUSION: Ethical judgements and decisions about research can be contradictory and be influenced by competing paradigms. Use of the concept of 'responsible advocacy', with due consideration to the principles of autonomy, beneficence, non-maleficence and justice, is recommended.

Impact of birthing practices on the breastfeeding dyad.

For breastfeeding to start and continue, the newborn must be able to suck, swallow, and breathe; the mother must be able and willing to let her infant breastfeed; and surroundings must support the biological unit: the mother-baby dyad. This article reviews how birth practices, including epidural anesthesia, cesarean surgery, forceps, and vacuum extraction, can affect the newborn's ability to feed, the mother's motivation and lactation capacity, and the mother-baby relationship.

The controversy about what constitutes safe and nurturant infant sleep environments.

In 1999, the U.S. Consumer Product Safety Commission stated that cribs provide the safest sleep environment for infants. Scientific data fails to support that statement and controversy continues in the scientific, medical, and parenting communities. Recent data demonstrate that cribs may represent the most unsafe sleep. This article seeks to inform health care professionals of the issues involved in the controversy and to offer guidelines for educating parents about safe and unsafe sleep practices.

Identification and characterization of non-pathogenic Fusarium oxysporum capable of increasing and decreasing Fusarium wilt severity.

Fusarium wilt of banana is a potentially devastating disease throughout the world. Options for control of the causal organism, Fusarium oxysporum f.sp. cubense (Foc) are limited. Suppressive soil sites have previously been identified where, despite the presence of Foc, Fusarium wilt does not develop. In order to understand some aspects of this disease suppression, endophytic Fusarium oxysporum isolates were obtained from banana roots. These isolates were genetically characterized and compared with an isolate of Fusarium oxysporum previously identified as being capable of suppressing Fusarium wilt of banana in glasshouse trials. Three additional isolates were selected for glasshouse trials to assess suppression of Fusarium wilt in two different cultivars of banana, Cavendish and Lady Finger. One isolate (BRIP 29089) was identified as a potential biocontrol organism, reducing the disease severity of Fusarium wilt in Lady Finger and Cavendish cultivars. Interestingly, one isolate (BRIP 45952) increased Fusarium wilt disease severity on Cavendish. The implications of an isolate of Fusarium oxysporum, non-pathogenic on banana, increasing disease severity and the potential role of non-pathogenic isolates of Fusarium oxysporum in disease complexes are discussed.

Hereditary dysautonomias: current knowledge and collaborations for the future.

The hereditary dysautonomias (H-Dys) are a large group of disorders that affect the autonomic nervous system. Research in the field of H-Dys is very challenging, because the disorders involve interdisciplinary, integrative, and "mind-body" connections. Recently, medical scientists, NIH/NINDS representatives, and several patient support groups gathered for the first time in order to discuss recent findings and future directions in the H-Dys field. The H-Dys workshop was instrumental in promoting interactions between basic science and clinical investigators. It also allowed attendees to have an opportunity to meet each other, understand the similarities between the various forms of dysautonomia, and experience the unique perspective offered by patients and their families. Future advances in H-Dys research will depend on a novel multi-system approach by investigators from different medical disciplines, and it is hoped that towards a common goal, novel "bench-to-bedside" therapeutics will be developed to improve the lives of, or even cure, patients suffering from dysautonomic syndromes.

Purification and structural characterization of the central hydrophobic domain of oleosin.

The oil bodies of rapeseeds contain a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with abundant structural alkaline proteins termed oleosins and some other minor proteins. Oleosins are unusual proteins because they contain a 70-80-residue uninterrupted nonpolar domain flanked by relatively polar C- and N-terminal domains. Although the hydrophilic N-terminal domain had been studied, the structural feature of the central hydrophobic domain remains unclear due to its high hydrophobicity. In the present study, we reported the generation, purification, and characterization of a 9-kDa central hydrophobic domain from rapeseed oleosin (19 kDa). The 9-kDa central hydrophobic domain was produced by selectively degrading the N and C termini with enzymes and then purifying the digest by SDS-PAGE and electroelution. We have also reconstituted the central domain into liposomes and synthetic oil bodies to determine the secondary structure of the domain using CD and Fourier transform infrared (FTIR) spectroscopy. The spectra obtained from CD and FTIR were analyzed with reference to structural information of the N-terminal domain and the full-length rapeseed oleosin. Both CD and FTIR analysis revealed that 50-63% of the domain was composed of beta-sheet structure. Detailed analysis of the FTIR spectra indicated that 80% of the beta-sheet structure, present in the central domain, was arranged in parallel to the intermolecular beta-sheet structure. Therefore, interactions between adjacent oleosin proteins would give rise to a stable beta-sheet structure that would extend around the surface of the seed oil bodies stabilizing them in emulsion systems. The strategies used in our present study are significant in that it could be generally used to study difficult proteins with different independent structural domains, especially with long hydrophobic domains.