A microbial supply chain for production of the anti-cancer drug vinblastine.

Monoterpene indole alkaloids (MIAs) are a diverse family of complex plant secondary metabolites with many medicinal properties, including the essential anti-cancer therapeutics vinblastine and vincristine1. As MIAs are difficult to chemically synthesize, the world's supply chain for vinblastine relies on low-yielding extraction and purification of the precursors vindoline and catharanthine from the plant Catharanthus roseus, which is then followed by simple in vitro chemical coupling and reduction to form vinblastine at an industrial scale2,3. Here, we demonstrate the de novo microbial biosynthesis of vindoline and catharanthine using a highly engineered yeast, and in vitro chemical coupling to vinblastine. The study showcases a very long biosynthetic pathway refactored into a microbial cell factory, including 30 enzymatic steps beyond the yeast native metabolites geranyl pyrophosphate and tryptophan to catharanthine and vindoline. In total, 56 genetic edits were performed, including expression of 34 heterologous genes from plants, as well as deletions, knock-downs and overexpression of ten yeast genes to improve precursor supplies towards de novo production of catharanthine and vindoline, from which semisynthesis to vinblastine occurs. As the vinblastine pathway is one of the longest MIA biosynthetic pathways, this study positions yeast as a scalable platform to produce more than 3,000 natural MIAs and a virtually infinite number of new-to-nature analogues.

Monocyte distribution width (MDW): study of reference values in blood donors.

OBJECTIVES: Monocyte distribution width (MDW) is a new biomarker used as an early indicator of sepsis (ESId). It is often aids in the identification of patients who may develop sepsis. This study aims to establish the MDW reference interval (RI) within the healthy population of blood donors using EDTA-K2 as anticoagulant. Many hospitals use this biomarker as a means of identifying patients who present to the hospital with sepsis. METHODS: A total of 274 samples obtained from healthy donors were analyzed. MDW measurements were taken within 2 h post-extraction. The RI was estimated using various statistical methodologies, including the recommended CLSI EP28-A3c guideline, non-parametric and robust methods, along with the Harrell-Davis bootstrap method applied to the entire sample. RESULTS: The RI estimated through non-parametric method was 14.77 CI90 % (14.36-14.97)-21.13 CI90 % (20.89-21.68); RI using the robust method was 15.64-19.05 and RI using the Harrell-Davis bootstrap method was 14.73 CI90 % (14.53-14.92)-21.14 CI90 % (20.88-21.40). CONCLUSIONS: Based on clinical applicability, we recommend utilizing the RI derived from the non-parametric method, aligning with the CLSI recommendations. Furthermore, we consider that our results can be taken as a reference in other laboratories that serve a population similar to our study cohort.

Radiological insights into rheumatoid meningitis - a rare central nervous system manifestation of rheumatoid arthritis: a retrospective review of six cases.

BACKGROUND: The objective is to analyze and review the clinical, laboratory, and neuroimaging characteristics of rheumatoid meningitis (RM) in six patients with known rheumatoid arthritis (RA). METHODS: We performed a retrospective review of patients diagnosed with RM from August 2012 to June 2023. To identify the cases, we used medical term search engines and the hospital´s radiology case database. Clinical information and laboratory findings were gathered from the medical records. A neuroradiologist with five years of experience reviewed and analyzed the RM to determine the characteristics findings of RM. RESULTS: Six patients with RM are included. Seizures along with headaches were among the clinical signs that were documented. All the patients had high levels of rheumatoid factor (RF) and anti-cyclic citrullinated peptides (ACPA) in the peripheral blood. Biopsy in two cases confirmed typical rheumatoid nodules. Leptomeningeal enhancement was found bilaterally in all cases and was predominantly found in the frontoparietal region. "Mismatch DWI/FLAIR" was found in five patients. Bilateral subdural collections could be found in two patients. Brain PET scan revealed increased metabolism in two cases. CONCLUSION: Rheumatoid meningitis is a rare complication of rheumatoid arthritis (RA) with challenging clinical diagnosis due to non-specific symptoms. This study highlights the importance of MR in detecting characteristic neuroimaging patterns, including "mismatch DWI/FLAIR", to aid in early diagnosis. Increased awareness of this condition may facilitate timely intervention and improve prognosis. These results still need to be verified by large studies.

Defining a competency framework for health and social professionals to promote healthy aging throughout the lifespan: an international Delphi study.

The promotion of healthy aging has become a priority in most parts of the world and should be promoted at all ages. However, the baseline training of health and social professionals is currently not adequately tailored to these challenges. This paper reports the results of a Delphi study conducted to reach expert agreement about health and social professionals' competencies to promote healthy aging throughout the lifespan within the SIENHA project. Materials and methods: This study was developed following the CREDES standards. The initial version of the competence framework was based on the results of a scoping review and following the CanMEDS model. The expert panel consisted of a purposive sample of twenty-two experts in healthy aging with diverse academic and clinical backgrounds, fields and years of expertise from seven European countries. Agreement was reached after three rounds. The final framework consisted of a set of 18 key competencies and 80 enabling competencies distributed across six domains. The SIENHA competence framework for healthy aging may help students and educators enrich their learning and the academic content of their subjects and/or programs and incentivize innovation.

The VR leadless pacemaker: Results of an expert panel using the RAND/UCLA method.

BACKGROUND: Leadless pacemakers were developed to reduce complications associated with transvenous pacemaker implantation and long-term follow-up. Existing international guidelines lack detailed instructions on patients suitable for leadless pacemaker implantation. Our aim was to develop a consensus document that provides medical guidance for all health professionals involved in the indication and implantation of Transcatheter Pacing System single-chamber device (VR leadless) pacemakers for patients with atrial fibrillation or in sinus rhythm. METHODS: A panel of experts, including interventional and non-interventional cardiologists, used the Research ANd Development/University of California at Los Angeles (RAND/UCLA) method to rate the appropriateness of leadless pacemaker implantation for 64 scenarios in patients with atrial fibrillation and 192 scenarios in sinus rhythm. The scenarios were rated individually and again during a moderated group session. Median ratings and level of agreement were calculated to classify each scenario as appropriate, inappropriate, or questionable. RESULTS: This consensus statement, based on available literature and the experts' opinions, summarizes recommendations for standardizing and optimizing leadless pacemaker implantation. The limitation for vascular access via the superior vena cava was the most influential variable when indicating leadless pacemaker implantation in both patients with atrial fibrillation and patients in sinus rhythm. CONCLUSIONS: Life expectancy, risk of infection, prosthetic valve, left ventricular ejection fraction (LVEF), limitation for vascular access via the superior vena cava, and mobility and exercise capacity determine who is advised to undergo VR leadless pacemaker implantation. More prospective studies are needed to optimize existing recommendations.

Global warming modifies the seasonal distribution of clutches on a Mediterranean great tit population.

Global warming has multiple effects on phenological events on a wide range of plants and animals. Specifically, many bird species have advanced the start of their breeding season, which could also imply an extension in its duration and also a change in the distribution of clutches throughout the breeding season. We have tested whether this occurred in a population of Great Tits (Parus major), in Sagunto (eastern Spain). The increase of March temperatures between 1986 and 2019 was related to an advancement of the breeding season. Although the breeding season was longer in years with higher June temperatures, the length did not show a temporal trend throughout the study period. The clutches were more concentrated at the beginning of the season (increase in the skewness), while the kurtosis ("tailedness" of the distribution) or the modality did not change significantly. Finally, the number of "equally good months" for breeding (a combined measure of length and distribution) has not changed throughout the years. Overall, all these phenological changes probably caused the observed increased proportion of pairs laying two clutches per year. It remains to be studied whether this increase in reproductive effort has positive or negative impact on fitness.

Aortoesophageal fistula as a fatal complication of esophageal perforation.

Aortoesophageal fistula is a very rare cause of upper gastrointestinal bleeding, however its interest lies in the high mortality rate associated with it. Due to this, early diagnosis and treatment of this entity is essential to increase survival. The typical symptoms known as the Chiari´s triad are only present in 45% of reported cases. We present the case of a patient with upper gastrointestinal bleeding due to an aortoesophageal fistula as well as the importance of endoscopic use for its differential diagnosis.

Prevalence of drug use before and during imprisonment in seven European countries (2014-2018).

Substance use is a global phenomenon that is particularly affecting the prison population. This study aimed to describe the prevalence of drug use among people in prison before and during incarceration in seven European countries and to compare it with the prevalence in the general population. Individual data collection was carried out between 2014 and 2018 with a model European Questionnaire on Drug Use among people in prison. A total of 12,918 people living in prison filled in the survey. People in prison report higher level of drug use when compared with the general population and the use of drug inside prison exist, although at lower levels when compared with predetention. Prisons can represent a point of access to engage individuals who use drugs in interventions that address drug use and risk factors related to both drug use and imprisonment.

Insulin-degrading enzyme ablation in mouse pancreatic alpha cells triggers cell proliferation, hyperplasia and glucagon secretion dysregulation.

AIMS/HYPOTHESIS: Type 2 diabetes is characterised by hyperglucagonaemia and perturbed function of pancreatic glucagon-secreting alpha cells but the molecular mechanisms contributing to these phenotypes are poorly understood. Insulin-degrading enzyme (IDE) is present within all islet cells, mostly in alpha cells, in both mice and humans. Furthermore, IDE can degrade glucagon as well as insulin, suggesting that IDE may play an important role in alpha cell function in vivo. METHODS: We have generated and characterised a novel mouse model with alpha cell-specific deletion of Ide, the A-IDE-KO mouse line. Glucose metabolism and glucagon secretion in vivo was characterised; isolated islets were tested for glucagon and insulin secretion; alpha cell mass, alpha cell proliferation and α-synuclein levels were determined in pancreas sections by immunostaining. RESULTS: Targeted deletion of Ide exclusively in alpha cells triggers hyperglucagonaemia and alpha cell hyperplasia, resulting in elevated constitutive glucagon secretion. The hyperglucagonaemia is attributable in part to dysregulation of glucagon secretion, specifically an impaired ability of IDE-deficient alpha cells to suppress glucagon release in the presence of high glucose or insulin. IDE deficiency also leads to α-synuclein aggregation in alpha cells, which may contribute to impaired glucagon secretion via cytoskeletal dysfunction. We showed further that IDE deficiency triggers impairments in cilia formation, inducing alpha cell hyperplasia and possibly also contributing to dysregulated glucagon secretion and hyperglucagonaemia. CONCLUSIONS/INTERPRETATION: We propose that loss of IDE function in alpha cells contributes to hyperglucagonaemia in type 2 diabetes.

Emotional experience of the diagnostic process of a rare disease and the perception of support systems: A scoping review.

AIMS AND OBJECTIVE: To explore the experience of adult patients and adult patients' families, and their perception of the support systems received during the diagnostic process of rare diseases. BACKGROUND: There are about 7,000 rare diseases that affect 7% of the world's population. Rare diseases are often underdiagnosed. This has been reported to have deleterious physical and psychological consequences in both the patients and their families, especially when institutional support during this process is low. DESIGN: A scoping review was carried out following the 6-phase model proposed by Arksey & O'Malley and Levac et al., including the consultation phase in which patients diagnosed with rare diseases were interviewed to seek their views on the bibliographic evidence reviewed and their experience during the diagnostic process. METHODS: The databases consulted were PubMed, CINAHL, Web of Science, SCOPUS, Cochrane Library, PsycINFO, OpenGrey, ProQuest Dissertations and Theses Global. They were explored from inception-July 2020, and qualitative, quantitative and mixed method studies were included. The Mixed Methods Appraisal Tool was used for the critical evaluation of the articles. The review was based on the guidance in the PRISMA-ScR statement. RESULTS: The initial search identified 2,350 articles, of which 20 fully met the inclusion criteria and were therefore reviewed. In this analysis appeared two dimensions: internal factors: emotional aspects, and external factors: resources and support systems. RELEVANCE TO CLINICAL: This review provides evidence on the emotional impact of the diagnostic process and during the communication phase of the definitive diagnosis. Health systems and professionals must be strengthened in order to improve the information, training and resources. Nurses can play a key role in coordinating communication and follow-up of those affected.

ImmunoCAP ISAC in food allergy diagnosis: a systematic review of diagnostic test accuracy.

OBJECTIVE: To assess the diagnostic test accuracy of the component-resolved diagnosis device ImmunoCAP ISAC, compared with oral food challenge. DESIGN: Systematic review reported according to the PRISMA-DTA recommendations. DATA SOURCES: Medline, Embase and Cochrane Library databases were searched from inception to May 2019 and updated in March 2021. ELIGIBILITY CRITERIA FOR SELECTING STUDIES: We included diagnostic test accuracy studies comparing ISAC component results as the index test with oral food challenge as the reference test, in people of any age suspected of IgE mediated food allergy to milk, egg, peanut, shrimp, hake, apple, peach, kiwi, melon, walnut, hazelnut, wheat or pineapple. Risk of bias was evaluated using the QUADAS-2 tool. RESULTS: We screened 799 titles and included 11 studies - seven prospective and two retrospective cohort studies, two case-control studies. Included studies evaluated IgE to Gald1 (three studies, 300 participants, 140 with egg allergy), Bosd5 (three studies, 242 participants, 146 with milk allergy) and Arah1 or 2 (seven studies, 546 participants, 346 with peanut allergy). No studies were identified for other ISAC components. Risk of bias was high or unclear mainly due to inadequate blinding. Applicability was of high or unclear concern due to unclear thresholds, inappropriate exclusions and variable populations. Gald1 sensitivity ranged from 58 to 84%, specificity 87%-97%. Bosd5 sensitivity 24%-40%, specificity 94%-95%. Arah1 sensitivity 45%-91%, specificity 41%-93%. Arah2 sensitivity 70%-94%, specificity 75%-95%. CONCLUSIONS: Diagnostic test accuracy information for ISAC components was only available for milk, egg and peanut. Specificity is generally higher than sensitivity, which contrasts with the performance of skin prick and standard specific IgE tests for diagnosing food allergy. Higher quality information is needed to determine the clinical utility of ISAC for food allergy diagnosis. SYSTEMATIC REVIEW REGISTRATION: Not registered.

A comparison of European countries FBDG in the light of their contribution to tackle diet-related health inequalities.

BACKGROUND: The purpose of this article is to report on a comparative analysis of the official food-based dietary guidelines (FBDG) that were applicable in 2015 in 25 EU Member States. We assess FBDG in relation to the main guidelines established by the FAO/WHO, the EURODIET project and the EFSA, with a particular focus on identifying strengths and limitations of current FBDG in Europe towards addressing diet-related health inequalities. METHODS: This is a review research, in which a mixed-methods sequenced procedure was utilized. In each EU country key informants, including sociologists, economists, dietitians and nutritionists were asked to provide data regarding: (i) current dietary guidelines and national health priorities, (ii) model of health promotion currently available, (iii) results of the latest food consumption survey. All documents were reviewed by the coordinating team. Full data were analysed by two nutritionists, using a tabulated sheet to organize and compare the results. RESULTS: While all countries have national FBDG, the level of detail and quality varies substantially with regard to: time of last update; availability of recommendations for specific target groups; specification of frequency and portion size; the graphical representation; recommended amounts and limits of foods consumed; and recommendations regarding physical activity. CONCLUSIONS: European countries have great opportunities to improve FBDG to better serve Public Health policy through a more consistent foundation of how these guidelines are developed, the inclusion of different population subgroups as a target for recommendations and the implementation of monitoring systems.

Comparison of the efficacy of 2 sedative protocols in pediatric dogs undergoing brainstem auditory-evoked response testing.

This study compared the quality of sedation with dexmedetomidine or alfaxalone during brainstem auditory-evoked response (BAER) tests in 6- to 17-week-old dogs. This was a prospective, randomized clinical study involving 19 client-owned pediatric dogs of breeds with reported congenital deafness. Group A (GA) received alfaxalone, 2 mg/kg body weight (BW) (n = 9) and group D (GD) dexmedetomidine, 0.005 mg/kg BW, and postprocedure antagonism with atipamezole (n = 10) intramuscularly. Time from injection to sedation, duration of sedation, sedation scores, need for re-dosing, rectal temperature, pulse and respiratory rate were recorded at baseline, before and after the BAER test, and once recovered from sedation. Pulse rate was significantly lower in GD (P = 0.004) and the number of re-dosing was significantly higher in GA (P = 0.011). Both sedation protocols allowed good quality BAER test recordings in pediatric dogs. Sedation with dexmedetomidine required less re-dosing, whereas alfaxalone maintained more physiological pulse rates.

Comparaison de l'efficacité de 2 protocoles de sédation chez des chiens pédiatriques soumis à un test potentiel évoqué auditif. L'étude vise à comparer deux protocoles de sédation à base de la dexmédétomidine et de l'alfaxalone pour la réalisation de test de potentiels évoqués auditifs (PEA) chez les chiens âgés de 6 à 17 semaines. Il s'agit d'une étude clinique prospective, randomisée, incluant 19 chiens pédiatriques de propriétaire, appartenant à des races prédisposées à la surdité congénitale. Les groupe A (GA) a reçu de l'alfaxalone (2 mg/kg) (n = 9), ceux du groupe D (GD) de la dexmédétomidine (0,005 mg/kg) (n = 10), en intramusculaire. Ont été relevés : temps d'action, durée de sédation, scores de sédation, nombre de doses, température, pouls et fréquence respiratoire; au repos, avant et après le test PEA. Des différences statistiquement significatives ont été trouvées dans la fréquence du pouls, étant plus bas pour GD (P = 0,004) alors que le nombre de doses utilisées, étant supérieurs parmi GA (P = 0,011). Trois chiens avaient une surdité unilatérale. Les deux protocoles de sédation ont permis des enregistrements de bonne qualité. La sédation avec la dexmédétomidine a nécessité moins de redosage; cependant, l'alfaxalone induit un pouls cardiaque plus proche des valeurs physiologiques chez les jeunes chiens testés.(Traduit par les auteurs).

Are Pediatric Critical Personnel Satisfied With Their Lives? Prediction of Satisfaction With Life From Burnout, Posttraumatic Stress, and Posttraumatic Growth, and Comparison With Noncritical Pediatric Staff.

OBJECTIVES: Staff in PICUs shows high burnout, posttraumatic stress disorder symptoms, and posttraumatic growth levels. However, their levels of satisfaction with life and how positive and negative posttrauma outcomes relate to each other and contribute to predict satisfaction with life remain unknown. Thus, we attempted to explore these aspects and to compare the findings with data from pediatric professionals working in noncritical units. DESIGN: This is an observational multicentric, cross-sectional study. SETTING: The PICU of nine hospitals in Spain, and other pediatric units in the same hospitals. SUBJECTS: Two hundred ninety-eight PICU workers (57 physicians, 177 nurses, and 64 nursing assistants) and 189 professionals working in noncritical pediatric units (53 physicians, 104 nurses, and 32 nursing assistants). INTERVENTION: Participants completed the Maslach Burnout Inventory, the Trauma Screening Questionnaire, the Posttraumatic Growth Inventory, and the Satisfaction With Life Scale. MEASUREMENTS AND MAIN RESULTS: Of PICU staff, 16.4% were very satisfied with their lives, 34.2% were satisfied, 34.6% showed average satisfaction with life, and 14.8% were below average. No differences were found between PICU and non-PICU workers. Women reported lower satisfaction with life than men, and physicians reported higher satisfaction with life than other professional groups. The correlation between posttraumatic stress disorder and posttraumatic growth was low, but significant and positive. According to the path analysis with latent variables, 20% of the variance satisfaction with life could be predicted from burnout, posttraumatic stress disorder symptoms, and posttraumatic growth. Higher distress was inversely associated to satisfaction with life, whereas posttraumatic growth contributed to higher satisfaction with life. CONCLUSIONS: Posttraumatic growth can moderate the negative effect of traumatic work-related experiences in satisfaction with life. PICU and non-PICU workers were equally satisfied with their lives. Positive and negative impact of work-related potentially traumatic events can coexist in the same person. Interventions aimed at reducing distress and fostering posttraumatic growth could impact in an improvement in pediatric health professionals' satisfaction with life.

Burnout and posttraumatic stress in paediatric critical care personnel: Prediction from resilience and coping styles.

INTRODUCTION: Our aims were (1) to explore the prevalence of burnout syndrome (BOS) and posttraumatic stress disorder (PTSD) in a sample of Spanish staff working in the paediatric intensive care unit (PICU) and compare these rates with a sample of general paediatric staff and (2) to explore how resilience, coping strategies, and professional and demographic variables influence BOS and PTSD. MATERIALS AND METHODS: This is a multicentre, cross-sectional study. Data were collected in the PICU and in other paediatric wards of nine hospitals. Participants consisted of 298 PICU staff members (57 physicians, 177 nurses, and 64 nursing assistants) and 189 professionals working in non-critical paediatric units (53 physicians, 104 nurses, and 32 nursing assistants). They completed the Brief Resilience Scale, the Coping Strategies Questionnaire for healthcare providers, the Maslach Burnout Inventory, and the Trauma Screening Questionnaire. RESULTS: Fifty-six percent of PICU working staff reported burnout in at least one dimension (36.20% scored over the cut-off for emotional exhaustion, 27.20% for depersonalisation, and 20.10% for low personal accomplishment), and 20.1% reported PTSD. There were no differences in burnout and PTSD scores between PICU and non-PICU staff members, either among physicians, nurses, or nursing assistants. Higher burnout and PTSD rates emerged after the death of a child and/or conflicts with patients/families or colleagues. Around 30% of the variance in BOS and PTSD is predicted by a frequent usage of the emotion-focused coping style and an infrequent usage of the problem-focused coping style. DISCUSSION AND CONCLUSIONS: Interventions to prevent and treat distress among paediatric staff members are needed and should be focused on: (i) promoting active emotional processing of traumatic events and encouraging positive thinking; (ii) developing a sense of detached concern; (iii) improving the ability to solve interpersonal conflicts, and (iv) providing adequate training in end-of-life care.

Inequalities in Health Services Usage in a National Health System Scheme: The Case of a Southern Social European Region.

BACKGROUND: Health services can reduce inequalities caused by other determinants of health or increase them due to the effect of the inverse care law-the principle that the availability of good quality care tends to vary inversely with the need for it in the population served. OBJECTIVE: The purpose of the research was to describe inequalities in the use of nursing services, medical services in primary care, specialist care, and services not fully covered by the Basque public health system in Spain. METHODS: A cross-sectional study of adults aged at least 25 years who completed the 2013 Basque Health Survey (N = 10,454) was conducted. Age-standardized prevalence and prevalence ratios for use of services that are covered and noncovered in the health system were computed. The association of health services usage with socioeconomic variables was estimated using a Poisson regression model with robust variance. The relative index of inequality (RII) was used to measure the magnitude of socioeconomic status inequalities in health service use. All analyses were carried out separately for men and women. RESULTS: Individuals with lower socioeconomic status were more likely to use primary care (RII = 0.87, 95% CI [0.79, 0.97]) and less likely to use specialist services (RII = 0.82, 95% CI [0.75, 0.89]). Across noncovered health services, inequalities between the highest and lowest social groups were significant in all cases and especially marked in men's use of physiotherapists (RII = 0.46, 95% CI [0.35, 0.61]) and podiatrists (RII = 0.24, 95%CI [0.15, 0.38]). DISCUSSION: There are significant inequalities in primary and specialist health service use based on individual socioeconomic status, particularly for services that are not provided free of charge within the existing health system. This suggests that health service systems that are not explicitly designed to provide universal access may actually amplify preexisting social and health inequalities within their target populations.

Social influences on adolescents' dietary behavior in Catalonia, Spain: A qualitative multiple-cases study from the perspective of social capital.

Adolescence has been referred to as the last best chance to prevent adult non-communicable diseases. Gaining further evidence on the psychosocial determinants of health behaviors, particularly the impact of peers, social networks and media on diet, is necessary to develop appropriate preventive strategies. Based on a multiple-cases study, our aim was to discuss the social influences on adolescents' dietary behavior from a social capital perspective. Participants were reached through four high-schools in different Catalan rural-urban and socioeconomic contexts. Our results confirm the different layout of social capital in the community, school, peers and family. In our sample, family and peers are the most influent sources of social capital in relation to dietary behaviors, inducing both protective and damaging effects.

Development and characterization of twelve microsatellite markers for Porphyra linearis Greville.

The genus Porphyra (and its sister genus Pyropia) contains important red algal species that are cultivated and/or harvested for human consumption, sustaining a billion-dollar aquaculture industry. A vast amount of research has been focused on species of this genus, including studies on genetics and genomics among other areas. Twelve novel microsatellite markers were developed here for Porphyra linearis. Markers were characterized using 32 individuals collected from four natural populations of P. linearis with total heterozygosity varying from 0.098 to 0.916. The number of alleles per locus ranged from 2 to 18. All markers showed cross amplification with Porphyra umbilicalis and/or Porphyra dioica. These polymorphic microsatellite markers are useful for investigating population genetic diversity and differentiation in P. linearis and may become useful for other genetic research on the reproductive biology of this important species.