Exploitation of microbial activities at low pH to enhance planetary health.

Awareness is growing that human health cannot be considered in isolation but is inextricably woven with the health of the environment in which we live. It is, however, under-recognized that the sustainability of human activities strongly relies on preserving the equilibrium of the microbial communities living in/on/around us. Microbial metabolic activities are instrumental for production, functionalization, processing, and preservation of food. For circular economy, microbial metabolism would be exploited to produce building blocks for the chemical industry, to achieve effective crop protection, agri-food waste revalorization, or biofuel production, as well as in bioremediation and bioaugmentation of contaminated areas. Low pH is undoubtedly a key physical-chemical parameter that needs to be considered for exploiting the powerful microbial metabolic arsenal. Deviation from optimal pH conditions has profound effects on shaping the microbial communities responsible for carrying out essential processes. Furthermore, novel strategies to combat contaminations and infections by pathogens rely on microbial-derived acidic molecules that suppress/inhibit their growth. Herein, we present the state-of-the-art of the knowledge on the impact of acidic pH in many applied areas and how this knowledge can guide us to use the immense arsenal of microbial metabolic activities for their more impactful exploitation in a Planetary Health perspective.

First Report of Alternaria alternata Causing Decay in Exported Sweet Basil During Freight from Israel to Europe.

Sweet basil (Ocimum basilicum) is an important spice herb grown in Israel for local markets and export. The crop is used as a fresh culinary herb or spice, and the essential oils are used in cosmetics and food flavorings. Due to increased demand, the production area of basil has increased in Israel. Postharvest losses due to fungal disease are a major economic concern for growers. In the summer of 2019, a leaf spot was observed in postharvest shipments of sweet basil destined for Europe; in late winter of 2022, leaf spots were observed on greenhouse-grown sweet basil. Fungal isolates from infected leaves were characterized by morphology in culture as Alternaria spp. PCR amplification of the Alternaria major allergen Alt a1, ITS, and gdp gene regions of the recovered isolates confirmed the presence of A. alternata, a common pathogen of numerous herbs and spice plants. In vitro growth tests demonstrated that 25°C was the optimum temperature for growth of the isolates. The isolates were tested for pathogenicity and found to infect a commonly grown cultivar of basil, cultivar Eli (previously cultivar Perrie). Foliar symptoms in pathogenicity tests were identical to those observed in commercial shipments and in the field, which completed Koch's postulates. Control of the nascent disease by applying fungicides to the plants may be necessary to reduce postharvest losses.

An overview of food lipids toward food lipidomics.

Increasing evidence regarding lipids' beneficial effects on human health has changed the common perception of consumers and dietary officials about the role(s) of food lipids in a healthy diet. However, lipids are a wide group of molecules with specific nutritional and bioactive properties. To understand their true nutritional and functional value, robust methods are needed for accurate identification and quantification. Specific analytical strategies are crucial to target specific classes, especially the ones present in trace amounts. Finding a unique and comprehensive methodology to cover the full lipidome of each foodstuff is still a challenge. This review presents an overview of the lipids nutritionally relevant in foods and new trends in food lipid analysis for each type/class of lipids. Food lipid classes are described following the LipidMaps classification, fatty acids, endocannabinoids, waxes, C8 compounds, glycerophospholipids, glycerolipids (i.e., glycolipids, betaine lipids, and triglycerides), sphingolipids, sterols, sercosterols (vitamin D), isoprenoids (i.e., carotenoids and retinoids (vitamin A)), quinones (i.e., coenzyme Q, vitamin K, and vitamin E), terpenes, oxidized lipids, and oxylipin are highlighted. The uniqueness of each food group: oil-, protein-, and starch-rich, as well as marine foods, fruits, and vegetables (water-rich) regarding its lipid composition, is included. The effect of cooking, food processing, and storage, in addition to the importance of lipidomics in food quality and authenticity, are also discussed. A critical review of challenges and future trends of the analytical approaches and computational methods in global food lipidomics as the basis to increase consumer awareness of the significant role of lipids in food quality and food security worldwide is presented.

Glucose levels are not the same for everyone: a real-world big data study evaluating fasting serum glucose levels by sex and age among children.

OBJECTIVES: Understanding the normal range of laboratory values as pertained to different age groups and males or females is paramount in health care delivery. We aimed to assess the distribution of morning fasting serum glucose levels by age and sex in the general population of children using a large-scale population-based cohort. METHODS: A retrospective study with real-world de-identified data from a large, state mandated health fund in Israel among children aged 2-18 years old between 2006 and 2019. Age, sex, and BMI differences in mean glucose levels were evaluated. RESULTS: Study included 130,170 venous blood samples from 117,411 children, 53.3 % were female. After adjusting for age boys had higher fasting serum glucose levels than girls, with a mean of 89.21 ± 8.66 mg/dL vs. 87.59 ± 8.35 (p<0.001) [4.95 ± 0.48 mmol/L vs. 4.86 ± 0.46]. Compared to the 15 to 18 year-olds (88.49 ± 7.63 mg/dL) [4.92 ± 0.42 mmol/L], 2 to 5 year-olds had lower glucose levels (84.19 ± 10.65, [4.68 ± 0.59] (p<0.001)), 11 to 14 year-olds had higher glucose (90.40 ± 7.42 [5.02 ± 0.41], (p<0.001)) and 6 to 10 year-olds showed no difference (88.45 ± 8.25) [4.91 ± 0.46]. 33.0 % (n=42,991) had a BMI percentile record the same year as their glucose test result. There was a weak yet significant positive association between blood glucose levels and BMI. CONCLUSIONS: Our large cohort indicates that boys have slightly higher fasting serum glucose levels than girls, as do adolescents compared to younger children. This finding is important for the delivery of adequate health care, screening for illness and avoiding unnecessary investigations and tests.

Lipid biomarkers for algal resistance to viral infection in the ocean.

Marine viruses play a key role in regulating phytoplankton populations, greatly affecting the biogeochemical cycling of major nutrients in the ocean. Resistance to viral infection has been reported for various phytoplankton species under laboratory conditions. Nevertheless, the occurrence of resistant cells in natural populations is underexplored due to the lack of sensitive tools to detect these rare phenotypes. Consequently, our current understanding of the ecological importance of resistance and its underlying mechanisms is limited. Here, we sought to identify lipid biomarkers for the resistance of the bloom-forming alga Emiliania huxleyi to its specific virus, E. huxleyi virus (EhV). By applying an untargeted lipidomics approach, we identified a group of glycosphingolipid (GSL) biomarkers that characterize resistant E. huxleyi strains and were thus termed resistance-specific GSLs (resGSLs). Further, we detected these lipid biomarkers in E. huxleyi isolates collected from induced E. huxleyi blooms and in samples collected during an open-ocean E. huxleyi bloom, indicating that resistant cells predominantly occur during the demise phase of the bloom. Last, we show that the GSL composition of E. huxleyi cultures that recover following infection and gain resistance to the virus resembles that of resistant strains. These findings highlight the metabolic plasticity and coevolution of the GSL biosynthetic pathway and underscore its central part in this host-virus arms race.

Leukocytosis and C-Reactive Protein Levels as Indicators of Infection in Children With Diabetic Ketoacidosis.

BACKGROUND: Diabetic ketoacidosis (DKA) is one of the serious complications of type 1 diabetes mellitus and may be aggravated by infection. Diagnosing an infection in a patient with DKA is often complicated because of the overlap of symptoms and the presence of leukocytosis in both conditions. Reliable indicators for the diagnosis of bacterial infection in DKA may reduce unnecessary use of antibiotics and enable closer monitoring of patients at risk. METHODS: This is a retrospective study. The study cohort included 180 children and adolescents with type 1 diabetes mellitus who were admitted to the Pediatric Emergency Department at Shaare Zedek Medical Center and had blood test results. We compared white blood cell count, C-reactive protein (CRP) levels, blood glucose levels, pH, the degree of acidosis, and the incidence of infection in patients with and without DKA. RESULTS: The incidence of probable bacterial infection in the entire cohort was 13.9%: 15.7% in the DKA group and 7.5% in the non-DKA group ( P = 0.65). The incidence of leukocytosis was significantly higher in patients with DKA ( P = 0.0003), although this was not related to bacterial infection. The CRP levels were higher in the DKA group with infection than without infection, and this was statistically significant ( P = 0.008). CONCLUSIONS: Our findings suggest that leukocytosis in DKA is not a reliable indicator of concomitant bacterial infection. In contrast, CRP levels were not related to the DKA or degree of acidosis and were significantly higher in patients with infection within the DKA group, and are therefore a more reliable indicator of concomitant infection in these patients.

Genetic and biotechnological tools to identify breeding targets for improving postharvest quality and extending shelf life of peppers.

Improved postharvest storage is a major target for pepper-crop production. The three main components of postharvest improvement of pepper fruit are reducing water-loss rate, reducing chilling susceptibility, and increasing resistance to pathogens. To date, a small number of Quantitative Trait Locus (QTL) studies have been reported for reduced water loss and enhanced tolerance to chilling and anthracnose. More effort is needed to screen germplasm collections for accessions with improved postharvest traits. Molecular studies have enabled the identification of candidate genes conferring reduced susceptibility to chilling injury and pathogen infection in pepper fruit, and in related crops such as tomato - which may be implemented in pepper. Manipulation of the activity of these genes by genome editing can improve postharvest pepper quality.

A COST Action on microbial responses to low pH: Developing links and sharing resources across the academic-industrial divide.

We present work of our COST Action on "Understanding and exploiting the impacts of low pH on micro-organisms". First, we summarise a workshop held at the European Federation of Biotechnology meeting on Microbial Stress Responses (online in 2020) on "Industrial applications of low pH stress on microbial bio-based production", as an example of an initiative fostering links between pure and applied research. We report the outcomes of a small survey on the challenging topic of developing links between researchers working in academia and industry that show that, while people in different sectors strongly support such links, barriers remain that obstruct this process. We present the thoughts of an expert panel held as part of the workshop above, where people with experience of collaboration between academia and industry shared ideas on how to develop and maintain links. Access to relevant information is essential for research in all sectors, and because of this we have developed, as part of our COST Action goals, two resources for the free use of all researchers with interests in any aspects of microbial responses to low pH. These are (1) a comprehensive database of references in the literature on different aspects of acid stress responses in different bacterial and fungal species, and (2) a database of research expertise across our network. We invite the community of researchers working in this field to take advantage of these resources to identify relevant literature and opportunities for establishing collaborations.

Self-Cleaning Biomimetic Surfaces-The Effect of Microstructure and Hydrophobicity on Conidia Repellence.

Modification of surface structure for the promotion of food safety and health protection is a technology of interest among many industries. With this study, we aimed specifically to develop a tenable solution for the fabrication of self-cleaning biomimetic surface structures for agricultural applications such as post-harvest packing materials and greenhouse cover screens. Phytopathogenic fungi such as Botrytiscinerea are a major concern for agricultural systems. These molds are spread by airborne conidia that contaminate surfaces and infect plants and fresh produce, causing significant losses. The research examined the adhesive role of microstructures of natural and synthetic surfaces and assessed the feasibility of structured biomimetic surfaces to easily wash off fungal conidia. Soft lithography was used to create polydimethylsiloxane (PDMS) replications of Solanum lycopersicum (tomato) and Colocasia esculenta (elephant ear) leaves. Conidia of B. cinerea were applied to natural surfaces for a washing procedure and the ratios between applied and remaining conidia were compared using microscopy imaging. The obtained results confirmed the hypothesis that the dust-repellent C. esculenta leaves have a higher conidia-repellency compared to tomato leaves which are known for their high sensitivities to phytopathogenic molds. This study found that microstructure replication does not mimic conidia repellency found in nature and that conidia repellency is affected by a mix of parameters, including microstructure and hydrophobicity. To examine the effect of hydrophobicity, the study included measurements and analyses of apparent contact angles of natural and synthetic surfaces including activated (hydrophilic) surfaces. No correlation was found between the surface apparent contact angle and conidia repellency ability, demonstrating variation in washing capability correlated to microstructure and hydrophobicity. It was also found that a microscale sub-surface (tomato trichromes) had a high conidia-repelling capability, demonstrating an important role of non-superhydrophobic microstructures.

First Report of Fruit Rot of Sweet Pepper Caused by Cladosporium cladosporioides in Israel.

Sweet pepper (Capsicum annuum L.) is an important vegetable crop grown in Israel for local and export markets. Bell pepper 'Dinamo' and sweet bite pepper exhibiting fruit spotting were observed in postharvest storage of commercial farms in Giv'at Ko'ah (32°02'21″ N; 34°56'41″ E) and Arava (30°46'41″ N; 35°14'28″ E) in July, 2021. Fruit with lesions were sporadic at each facility, however, limited numbers of packaged boxes were discarded due to a high percentage of symptomatic fruit. The spots appeared as slightly sunken dark brown or black lesions, coated by a fine velvety layer of olive-grey conidia (Suppl. Fig. 1). A fungus was routinely isolated from the fruit onto potato dextrose agar amended with 50 ppm chloramphenicol (PDAC) and was tentatively identified as a Cladosporium spp. based on colony morphology and conidial appearance (Suppl. Fig. 1). Isolates (n = 6) of the fungus were cultured on malt extract agar (MEA) and PDA for morphological observation. On MEA, the pathogen sporulated profusely and conidia measured 4.5 (3 - 7) × 3.5 (2 - 4) µm and were smooth and globose or sub-globose. Ramoconidia had 0 or 1 septations and measured 13.4 (10 - 18) × 3.9 (3 - 6) µm. Conidiophores were solitary, straight or flexous, usually unbranched, and measured 60 - 350 µm. Both macro- and micronematous conidiophores were present. Hyphae were flat and immersed, olivaceous-grey, septate and rope-like. The fungus was slow growing, reaching 41.0 mm after 10 d incubation at 20°C. To confirm species identity, the isolates were grown in potato dextrose broth and DNA was extracted from the mycelium using MasterPure DNA Purification Kit (EpiCentre, Madison, WI, USA) following the manufacturer's instructions. The internal transcribed spacer (ITS) region was amplified using primers ITS 1 and ITS 4, and elongation factor-1 and actin gene regions with EF1-728F/986R and ACT-512F/783R, respectively (Carbone and Kohn, 1999). The PCR products were sequenced and a BLASTn search showed 99-100% similarity to Cladosporium cladosporioides (GenBank accession nos. MW255614.1, MK416093.1, MK306457.1), a common fungal species found worldwide as a plant pathogen and saphrophyte (Bensch et al., 2012). The ITS gene sequences for a representative isolate from each location were deposited in GenBank (accession nos. OK104139.1 and OL672241.1). To confirm pathogenicity, red bell pepper fruits (n = 8) were wounded with a sterile needle and a 12 µl drop of conidial suspension (4 x 105/ml) of each isolate was pipetted onto the wound. Control fruit received sterile water. The fruit were incubated at 100% RH in sealed containers at 20-22°C. After 7-10 d incubation small dark sunken spots developed, and by 14 d lesions formed with velvety conidial growth similar to the ones originally observed. The fungus was successfully isolated again from the lesions onto PDA and confirmed as C. cladosporioides based on colony and conidial morphology. No symptoms developed on fruit inoculated with sterile water. The pathogenicity test was repeated two times with similar results. C. cladosporioides is a cosmopolitan pathogen and has been cited causing disease on numerous crops worldwide (Bensch et al., 2012). This species was reported causing tomato leaf spot in Mexico (Robles-Yerena et al., 2019) and black mould of tomato fruit in Australia (Ma et al., 2020). Cladosporium herbarum was found causing fruit rot of bell pepper in the U.S. (Ramsey and Heiberg, 1952). Sweet peppers are widely grown in Israel, and have high economic value as an export crop. Control measures to limit losses from C. cladosporioides postharvest may be necessary if conditions are favorable for disease during prolonged storage and refrigerated transport.

Daytime or Edge-of-Daytime Intra-Canopy Illumination Improves the Fruit Set of Bell Pepper at Passive Conditions in the Winter.

Optimal light conditions ensure the availability of sufficient photosynthetic assimilates for supporting the survival and growth of fruit organs in crops. One of the growing uses of light-emitting diodes (LEDs) in horticulture is intra-canopy illumination or LED-interlighting, providing supplemental light for intensively cultivated crops directly within their canopies. Originally developed and applied in environmentally controlled greenhouses in northern latitude countries, this technique is nowadays also being tested and studied in other regions of the world such as the Mediterranean region. In the present work, we applied intra-canopy illumination for bell pepper grown in passive high tunnels in the Jordan Valley using a commercial LED product providing cool-white light. The study included testing of daytime ('LED-D') and edge-of-daytime ('LED-N') illumination, as well as a detailed characterization of fruit set and fruit survival throughout the growth season. We found that both light regimes significantly improved the fruit set and survival during winter, with some benefit of LED-N illumination. Notably, we found that western-facing plants of illuminated sections had a higher contribution toward the increased winter fruit set and spring yield than that of illuminated eastern-facing plants. Greater plant height and fresh weight of western-facing plants of the illuminated sections support the yield results. The differences likely reflect higher photosynthetic assimilation of western-facing plants as compared to eastern-facing ones, due to the higher daily light integral and higher canopy temperature of the former. This study provides important implications for the use of intra-canopy lighting for crops grown at passive winter conditions and exemplifies the significance of geographical positioning, opening additional avenues of investigation for optimization of its use for improving fruit yield under variable conditions.

Sucrose promotes stem branching through cytokinin.

Shoot branching is an important aspect of plant architecture because it substantially affects plant biology and agricultural performance. Sugars play an important role in the induction of shoot branching in several species, including potato (Solanum tuberosum L.). However, the mechanism by which sugars affect shoot branching remains mostly unknown. In the present study, we addressed this question using sugar-mediated induction of bud outgrowth in potato stems under etiolated conditions. Our results indicate that sucrose feeding to detached stems promotes the accumulation of cytokinin (CK), as well as the expression of vacuolar invertase (VInv), an enzyme that contributes to sugar sink strength. These effects of sucrose were suppressed by CK synthesis and perception inhibitors, while CK supplied to detached stems induced bud outgrowth and VInv activity in the absence of sucrose. CK-induced bud outgrowth was suppressed in vinv mutants, which we generated by genome editing. Altogether, our results identify a branching-promoting module, and suggest that sugar-induced lateral bud outgrowth is in part promoted by the induction of CK-mediated VInv activity.

Stress Hyperglycemia in a Pediatric Emergency Department Setting and its Relationship to the Underlying Disease and Outcome.

BACKGROUND: Stress hyperglycemia (SH) is a common finding in patients in pediatric emergency departments (PED) and has been related to increased morbidity and mortality. OBJECTIVES: To assess the incidence of SH among children visiting the PED. To identify which diseases predispose patients to SH and whether they indicate a worse outcome. METHODS: Data were collected retrospectively from the medical records of all children aged 0-18 years who visited the PED during the years 2010-2014 and who had a glucose level of ≥ 150 mg/dl. Data collected included age, gender, weight, blood glucose level, presence or absence of a pre-existing or a new diagnosis of diabetes mellitus, and previous treatment with medications affecting blood glucose levels or with intravenous fluids containing dextrose. Data were collected regarding hospitalization, duration of hospitalization, discharge diagnosis, and survival status. RESULTS: The study population included 1245 children with SH, which comprised 2.6% of all patients whose blood glucose level was measured in the PED during the study period. The mean age of children with SH was 49 months; 709 (56.9%) were male. The mean blood glucose level was 184 mg/dl. The rate of hospitalization was 57.8%. The mean duration of hospital stay was 5.6 days and mortality rate was 0.96%. The majority were diagnosed with a respiratory illness. CONCLUSIONS: SH is a common phenomenon among children evaluated in the PED and is associated with a high incidence of hospitalization. It may serve as an additional clinical indicator of disease severity.

The inter - Test variability of growth hormone stimulation tests and factors affecting this variability.

OBJECTIVE: To evaluate the variability of growth hormone stimulation tests results and factors affecting it in short children suspected of having growth hormone deficiency. DESIGN: The cohort included patients with short stature suspected of having growth hormone deficiency, and who underwent a second stimulation test, after the first stimulation test was positive. Testing was done at a single center from May 2014 to October 2017. Patients' weight, height, age, sex, stimulating agents and test results were recorded. RESULTS: The study population comprised 200 patients, 108 males and 92 females, average age 9.2 years (2.2-16.6 years). The average peak growth hormone was 5.2 µg/L and 7.8 µg/L in the first and second tests respectively and the concordance rate was 56.5%. The probability of a second positive test was increased if the peak growth hormone level in the first test was below 5 µg/L. In the second test, Clonidine and Glucagon led to higher peak growth levels than Arginine with averages of 9.02, 9.97 and 6.88 µg/L respectively. Younger children and children with higher BMI SDS only had lower peaks of growth hormone in the second test. The effect of height SDS on peak growth hormone levels was equivocal. CONCLUSION: The reproducibility rate of GH simulation tests in our study was low. A few factors may affect the peak levels of growth hormone in the second test, the most prominent being the peak of growth hormone in the first test.

The novel founder homozygous V225M mutation in the HSD17B3 gene causes aberrant splicing and XY-DSD.

PURPOSE: Mutations in the gene HSD17B3 encoding the 17-beta hydroxysteroid dehydrogenase 3 enzyme cause testosterone insufficiency leading to XY disorders of sex development. In this study the clinical and molecular characteristics of three patients from consanguineous families are elucidated. METHODS: We identified three patients from two unrelated families with XY DSD and a novel homozygous HSD17B3:c. 673G>A mutation. The effect of the mutation on splicing was determined in RNA extracted from the testis of one patient. RESULTS: Three patients presented at ages 0.1, 8 and 0.7 years with ambiguous genitalia and an XY Karyotype. Endocrine workup showed normal cortisol and mineralocorticoid levels with a low testosterone/androstenedione ratio. Whole-exome sequencing, carried out in the first family, revealed a homozygous novel mutation in the HSD17B3 gene: c. 673G>A, p. V225M. The same mutation was found by Sanger sequencing in the third unrelated patient. Haplotype analysis of a 4 Mb region surrounding the HSD17B3 gene on chromosome 9 revealed that the mutation resides on the same allele in all three patients. The mutation, being the first nucleic acid on exon 10, affects splicing and causes exon 10 skipping in one of our patients' testes. CONCLUSION: The novel homozygous c. 673G>A, p. V225M mutation in the 17HSDB3 gene is likely a founder mutation and causes severe XY-DSD. It changes a conserved amino acid residue, and also alters 17HSDB3 gene transcription by causing skipping of exon 10, thereby contributing to an imbalance in the relevant protein isoforms and consequently, significant decreased 17HDSB3 enzymatic activity.

Sugar-regulated susceptibility of tomato fruit to Colletotrichum and Penicillium requires differential mechanisms of pathogenicity and fruit responses.

Colletotrichum gloeosporioides and Penicillium expansum cause postharvest diseases in tropical and deciduous fruit. During colonization, C. gloeosporioides and P. expansum secrete ammonia in hosts with low sugar content (LowSC) and gluconic acid in hosts with high sugar content (HighSC), respectively, as a mechanism to modulate enhanced pathogenicity. We studied the pathogens interactions with tomato lines of similar genetic background but differing in their sugar content. Colletotrichum gloeosporioides showed enhanced colonization of the LowSC line with differential expression response of 15% of its genes including enhanced relative expression of glycosyl hydrolases, glucanase and MFS-transporter genes. Enhanced colonization of P. expansum occurred in the HighSC line, accompanied by an increase in carbohydrate metabolic processes mainly phosphoenolpyruvate carboxykinase, and only 4% of differentially expressed genes. Gene response of the two host lines strongly differed depending on the sugar level. Limited colonization of HighSC line by C. gloeosporioides was accompanied by a marked alteration of gene expression compared the LowSC response to the same pathogen; while colonization by P. expansum resulted in a similar response of the two different hosts. We suggest that this differential pattern of fungal/host responses may be the basis for the differential of host range of both pathogens in nature.

How rootstock/scion combinations affect watermelon fruit quality after harvest?

Grafting of vegetable seedlings is a unique horticultural technology, practiced for more than five decades, aiming to overcome problems associated with intensive cultivation on limited arable land. Grafting can protect vegetables against soil-borne diseases and nematodes; against abiotic stresses such as high or low temperatures, salinity, drought or excessive soil-water content; and against elevated soil concentrations of heavy metals and organic pollutants. Watermelon is one of the most popular vegetables to be grafted, and more than 90% of the plants worldwide are commercially grafted. This mini review aims to summarize the latest available information about the effects of rootstock/scion combinations with respect to enhancing or impairing watermelon fruit-quality. A better understand of the influence of rootstock/scion compatibility or incompatibility on fruit-quality parameters will facilitate decision-making by growers and direct breeding programs to produce high-quality grafted fruits in a cost-effective manner. © 2020 Society of Chemical Industry.

Primary Ovarian Insufficiency Nationwide Incidence Rate and Etiology Among Israeli Adolescents.

PURPOSE: The aim of the study was to estimate the current incidence and the distribution of etiologies of primary ovarian insufficiency (POI) in a nationwide study. The prevalence of POI in young adult women has recently increased, but the data cited for adolescents are more than three decades old. METHODS: Data regarding females aged <21 years diagnosed with POI during the years 2000-2016 were collected from all the pediatric endocrinology units in Israel. POI was defined by at least 4 months of amenorrhea in association with menopausal levels of follicle-stimulating hormone. Iatrogenic cases were excluded. RESULTS: For the 130 females aged <21 years included in the study, the distribution of POI etiologies was Turner syndrome/mosaicism in 56 (43%), idiopathic in 35 (27%), and other (developmental, genetic, metabolic, adrenal, and autoimmune) in 39 (30%) females. During the years 2009-2016, compared with 2000-2008, the incidence rate of new POI diagnoses per 100,000 person-years doubled (4.5 vs. 2.0; p value <.0001), and incidence rates of idiopathic and other etiologies increased by 2.6 (p value = .008) and 3.0 (p value = .002), respectively. In contrast, the incidence of Turner syndrome was constant (p value = .2). In the age group of 15-21 years, the current incidence of non-Turner POI in adolescents is one per 100,000 person-years. CONCLUSIONS: In this nationwide study, the incidence rate of POI in youth aged <21 years was one tenth of the rate that is commonly cited. A significant increase in the rate of POI in non-Turner females was observed over the last decade. Contributions of environmental and epigenetic factors should be studied.

Synergistic Inhibition of Mycotoxigenic Fungi and Mycotoxin Production by Combination of Pomegranate Peel Extract and Azole Fungicide.

Fungal plant pathogens cause considerable losses in yield and quality of field crops worldwide. In addition, under specific environmental conditions, many fungi, including such as some Fusarium and Aspergillus spp., are further able to produce mycotoxins while colonizing their host, which accumulate in human and animal tissues, posing a serious threat to consumer health. Extensive use of azole fungicides in crop protection stimulated the emergence of acquired azole resistance in some plant and human fungal pathogens. Combination treatments, which become popular in clinical practice, offer an alternative strategy for managing potentially resistant toxigenic fungi and reducing the required dosage of specific drugs. In the current study we tested the effect of pomegranate peel extract (PPE) on the growth and toxin production of the mycotoxigenic fungi Aspergillus flavus and Fusarium proliferatum, both alone and in combination with the azole fungicide prochloraz (PRZ). Using time-lapse microscopy and quantitative image analysis we demonstrate significant delay of conidial germination and hyphal elongation rate in both fungi following PPE treatment in combination with PRZ. Moreover, PPE treatment reduced aflatoxin production by A. flavus up to 97%, while a combined treatment with sub-inhibitory doses of PPE and PRZ resulted in complete inhibition of toxin production over a 72 h treatment. These findings were supported by qRT-PCR analysis, showing down-regulation of key genes involved in the aflatoxin biosynthetic pathway under combined PPE/PRZ treatment al low concentrations. Our results provide first evidence for synergistic effects between the commercial drug PRZ and natural compound PPE. Future application of these findings may allow to reduce the required dosage of PRZ, and possibly additional azole drugs, to inhibit mycotoxigenic fungi, ultimately reducing potential concerns over exposure to high doses of these potentially harmful fungicides.