Uncovering social and environmental factors that increase the burden of climate-sensitive diarrheal infections on children.

Climate-sensitive infectious diseases are an issue of growing concern due to global warming and the related increase in the incidence of extreme weather and climate events. Diarrhea, which is strongly associated with climatic factors, remains among the leading causes of child death globally, disproportionately affecting populations in low- and middle-income countries (LMICs). We use survey data for 51 LMICs between 2000 and 2019 in combination with gridded climate data to estimate the association between precipitation shocks and reported symptoms of diarrheal illness in young children. We account for differences in exposure risk by climate type and explore the modifying role of various social factors. We find that droughts are positively associated with diarrhea in the tropical savanna regions, particularly during the dry season and dry-to-wet and wet-to-dry transition seasons. In the humid subtropical regions, we find that heavy precipitation events are associated with increased risk of diarrhea during the dry season and the transition from dry-to-wet season. Our analysis of effect modifiers highlights certain social vulnerabilities that exacerbate these associations in the two climate zones and present opportunities for public health intervention. For example, we show that stool disposal practices, child feeding practices, and immunizing against the rotavirus modify the association between drought and diarrhea in the tropical savanna regions. In the humid subtropical regions, household's source of water and water disinfection practices modify the association between heavy precipitation and diarrhea. The evidence of effect modification varies depending on the type and duration of the precipitation shock.

Years of good life is a well-being indicator designed to serve research on sustainability.

Sustainable development (SD) as popularized by the Brundtland Commission and politically enshrined in the Sustainable Development Goals has been the explicit focus of sustainability science. While there is broad agreement that the trend of human well-being (W) over time should serve as a sustainability criterion, the literature so far has mostly addressed this in terms of its determinants rather than focusing on W itself. There is broad agreement that an indicator for W should have multiple constituents, clearly going beyond gross domestic product. Here, we propose a tailor-made indicator to serve precisely this purpose following a set of specified desiderata, including its applicability to flexibly defined subnational populations by gender, place of residence, ethnicity, and other relevant characteristics. The indicator, years of good life (YoGL), reflects the evident fact that in order to be able to enjoy any quality of life, one has to be alive and thus is primarily based on life expectancy. However, since mere survival is not considered good enough, life years are counted conditional on meeting minimum standards in two dimensions: the objective dimension of capable longevity (consisting of being out of absolute poverty and enjoying minimal levels of physical and cognitive health) and the subjective dimension of overall life satisfaction. We illustrate the calculation of this indicator for countries and subpopulations at different stages of development and with different degrees of data availability.

Essential childhood immunization in 43 low- and middle-income countries: Analysis of spatial trends and socioeconomic inequalities in vaccine coverage.

BACKGROUND: Globally, access to life-saving vaccines has improved considerably in the past 5 decades. However, progress has started to slow down and even reverse in recent years. Understanding subnational heterogeneities in essential child immunization will be critical for closing the global vaccination gap. METHODS AND FINDINGS: We use vaccination information for over 220,000 children across 1,366 administrative regions in 43 low- and middle-income countries (LMICs) from the most recent Demographic and Health Surveys. We estimate essential immunization coverage at the national and subnational levels and quantify socioeconomic inequalities in such coverage using adjusted concentration indices. Within- and between-country variations are summarized via the Theil index. We use local indicator of spatial association (LISA) statistics to identify clusters of administrative regions with high or low values. Finally, we estimate the number of missed vaccinations among children aged 15 to 35 months across all 43 countries and the types of vaccines most often missed. We show that national-level vaccination rates can conceal wide subnational heterogeneities. Large gaps in child immunization are found across West and Central Africa and in South Asia, particularly in regions of Angola, Chad, Nigeria, Guinea, and Afghanistan, where less than 10% of children are fully immunized. Furthermore, children living in these countries consistently lack all 4 basic vaccines included in the WHO's recommended schedule for young children. Across most countries, children from poorer households are less likely to be fully immunized. The main limitations include subnational estimates based on large administrative divisions for some countries and different periods of survey data collection. CONCLUSIONS: The identified heterogeneities in essential childhood immunization, especially given that some regions consistently are underserved for all basic vaccines, can be used to inform the design and implementation of localized intervention programs aimed at eliminating child suffering and deaths from existing and novel vaccine-preventable diseases.

Microgravimetric and Spectroscopic Analysis of Solid-Electrolyte Interphase Formation in Presence of Additives.

Electrochemical quartz crystal microbalance (EQCM) with damping monitoring is applied for real-time analysis of solid-electrolyte interphase (SEI) formation in diphenyl octyl phosphate (DPOP) and vinylene carbonate (VC) modified electrolytes. Fast SEI formation is observed for the DPOP containing electrolyte, whereas slow growth is detected in VC-modified and reference electrolytes. QCM measurements in a dry state show considerable reduction of the mass quantity for DPOP and reference samples and minor mass decrease for the SEI layer formed in the presence of VC. The results indicate that VC enhances SEI stability, whereas the addition of DPOP or no additive results in incorporation of loosely attached species, leadubg to SEI instability. Resonance frequency damping, Δw, and dissipation factor, D, are used for analyzing mechanical properties of the SEI layers. The apparent increase of Δw and D during SEI formation in presence of DPOP suggests a pronounced viscoelasticity of the layer. QCM results are compared with surface morphology and chemical composition, revealing excellent agreement of the applied characterization approaches.

Global structural changes of an ion channel during its gating are followed by ion mobility mass spectrometry.

Mechanosensitive ion channels are sensors probing membrane tension in all species; despite their importance and vital role in many cell functions, their gating mechanism remains to be elucidated. Here, we determined the conditions for releasing intact mechanosensitive channel of large conductance (MscL) proteins from their detergents in the gas phase using native ion mobility-mass spectrometry (IM-MS). By using IM-MS, we could detect the native mass of MscL from Escherichia coli, determine various global structural changes during its gating by measuring the rotationally averaged collision cross-sections, and show that it can function in the absence of a lipid bilayer. We could detect global conformational changes during MscL gating as small as 3%. Our findings will allow studying native structure of many other membrane proteins.

The activation mode of the mechanosensitive ion channel, MscL, by lysophosphatidylcholine differs from tension-induced gating.

One of the best-studied mechanosensitive channels is the mechanosensitive channel of large conductance (MscL). MscL senses tension in the membrane evoked by an osmotic down shock and directly couples it to large conformational changes leading to the opening of the channel. Spectroscopic techniques offer unique possibilities to monitor these conformational changes if it were possible to generate tension in the lipid bilayer, the native environment of MscL, during the measurements. To this end, asymmetric insertion of l-α-lysophosphatidylcholine (LPC) into the lipid bilayer has been effective; however, how LPC activates MscL is not fully understood. Here, the effects of LPC on tension-sensitive mutants of a bacterial MscL and on MscL homologs with different tension sensitivities are reported, leading to the conclusion that the mode of action of LPC is different from that of applied tension. Our results imply that LPC shifts the free energy of gating by interfering with MscL-membrane coupling. Furthermore, we demonstrate that the fine-tuned addition of LPC can be used for controlled activation of MscL in spectroscopic studies.

Study of light-induced MscL gating by EPR spectroscopy.

A number of techniques developed to investigate protein structure and function depend on chemically modifying and/or labeling of proteins. However, in the case of homooligomeric proteins, the presence of multiple identical subunits obstructs the introduction of residue-specific labels to only one or several subunits, selectively. Here, in order to study the initial conformational changes of a homopentameric mechanosensitive ion channel during its gating, we developed a method for labeling a defined number of subunits of the channel with two different cysteine-specific compounds simultaneously. The first one is a light-sensitive channel activator that determines the degree of openness of the ion channel upon irradiation. The second one is a spin label, containing an unpaired electron, which allows following the resulting structural changes upon channel gating by electron paramagnetic resonance spectroscopy. With this method, we could open MscL into different sub-open states. As the number of light switches per channel increased, the intersubunit spin-spin interactions became less, indicating changes in intersubunit proximities and opening of the channel. The ability of controlled activation of MscL into different open states with a noninvasive trigger and following the resulting conformational changes by spectroscopy will pave the way for detailed spectroscopic studies in the area of mechanosensation.

Temperature-related neonatal deaths attributable to climate change in 29 low- and middle-income countries.

Exposure to high and low ambient temperatures increases the risk of neonatal mortality, but the contribution of climate change to temperature-related neonatal deaths is unknown. We use Demographic and Health Survey (DHS) data (n = 40,073) from 29 low- and middle-income countries to estimate the temperature-related burden of neonatal deaths between 2001 and 2019 that is attributable to climate change. We find that across all countries, 4.3% of neonatal deaths were associated with non-optimal temperatures. Climate change was responsible for 32% (range: 19-79%) of heat-related neonatal deaths, while reducing the respective cold-related burden by 30% (range: 10-63%). Climate change has impacted temperature-related neonatal deaths in all study countries, with most pronounced climate-induced losses from increased heat and gains from decreased cold observed in countries in sub-Saharan Africa. Future increases in global mean temperatures are expected to exacerbate the heat-related burden, which calls for ambitious mitigation and adaptation measures to safeguard the health of newborns.

Implications of climate-related disasters on refugees' health: A case study of resettled Syrian and Iraqi refugees in San Diego, California.

Background: Climate change disproportionately harms people of color and low-income communities. Despite their unprecedented numbers, being constantly on the move, and suffering extreme social vulnerability, almost nothing is known about the impact of climate change on the health of refugees. This study uses state-of-the-art mixed methods to examine the differential susceptibility of climate-sensitive exposures and environmental exposures among refugees and their links to perceived health after resettlement. Methods: Arabic-speaking refugees (N=67) from Iraq and Syria previously diagnosed with hypertension who resettled in California were recruited from a community center. Semi-structured interviews were conducted to explore participant's understanding of the impact of climate on health. Survey data were collected to inquire regarding participant's refugee journeys prior to resettlement in the US. Survey data on climate-related disasters was retrospectively geo-referenced through the Emergency Events Database (EM-DAT). Qualitative data was analyzed using inductive thematic analysis. Results: All participants stayed in at least one temporary resettlement country during their migration journey and 12% has stayed in refugee camps. The most popular resettlement sites were Turkey (most disaster-prone in the region due to frequent floods and earthquakes) and Jordan (one of the most extreme water-scarce globally). Participants reported harsh weather conditions during their migratory journeys including extreme cold in Turkey and extreme heat in Jordan. Many participants noted their exposure to dust throughout their travels, and an inability to deal with harsh weather conditions due to financial insecurity. Participants did not link their diagnosis of hypertension to their experience of extreme weather and would only link it to their exposure to stress from war. Participants did note poorer mental health due to poor weather conditions and a challenge adjusting to the climate conditions in different countries. Few participants reported residing in a refugee camp and described it as ill-equipped for the challenges of climate hazards. Conclusion: This study reveals the links between structural drivers of climate change and health inequities for refugee populations. Refugees are highly vulnerable to climate-sensitive exposures but remain not fully aware of the potential links between these exposures and health. Learnings from this study will inform clinical and public health interventions, and policies to close the climate gap without leaving this vulnerable population behind.

Saharan Dust and Childhood Respiratory Symptoms in Benin.

Mineral dust is one of the largest natural constituents of coarse particulate matter (PM10). Most of these dust emissions originate from northern Africa, and several hundred tera-grams of dust are emitted annually from this region. Previous evidence has linked dust PM10 to adverse respiratory outcomes in children. However, most of these studies have been from high-income countries (HICs) or examined dust from other regions of the world, mainly Asia. Evidence from low-to-middle-income countries (LMICs) in Africa is scarce. Respiratory infections are one of the leading causes of under-five mortality across the globe. However, there is a poignant disparity in studies examining these outcomes in children in the region where most dust is emitted. This study linked remotely sensed satellite data to a nationally representative survey to examine acute exposure to dust in children living in Benin using a time-stratified case-crossover analysis. We identified acute effects of exposure to dust and increased risk of cough in children under five. The effect of increased risk is strongest within two weeks of exposure and dissipates by four weeks. Children living in rural areas and households with lower income had a greater risk of adverse respiratory outcomes when exposed to dust. We could elucidate the specific period and conditions of increased risk for respiratory problems in children living in Benin.

Precipitation variability and risk of infectious disease in children under 5 years for 32 countries: a global analysis using Demographic and Health Survey data.

BACKGROUND: Precipitation variability is a potentially important driver of infectious diseases that are leading causes of child morbidity and mortality worldwide. Disentangling the links between precipitation variability and disease risk is crucial in a changing climate. We aimed to investigate the links between precipitation variability and reported symptoms of infectious disease (cough, fever, and diarrhoea) in children younger than 5 years. METHODS: We used nationally representative survey data collected between 2014 and 2019 from Demographic and Health Survey (DHS) surveys for 32 low-income to middle-income countries in combination with high-resolution precipitation data (via the Climate Hazards Group InfraRed Precipitation with Station dataset). We only included DHS data for which interview dates and GPS coordinates (latitude and longitude) of household clusters were available. We used a regression modelling approach to assess the relationship between different precipitation variability measures and infectious disease symptoms (cough, fever, and diarrhoea), and explored the effect modification of different climate zones and disease susceptibility factors. FINDINGS: Our global analysis showed that anomalously wet conditions increase the risk of cough, fever, and diarrhoea symptoms in humid, subtropical regions. These health risks also increased in tropical savanna regions as a result of anomalously dry conditions. Our analysis of susceptibility factors suggests that unimproved sanitation and unsafe drinking water sources are exacerbating these effects, particularly for rural populations and in drought-prone areas in tropical savanna. INTERPRETATION: Weather shifts can affect the survival and transmission of pathogens that are particularly harmful to young children. As our findings show, the health burden of climate-sensitive infectious diseases can be substantial and is likely to fall on populations that are already among the most disadvantaged, including households living in remote rural areas and those lacking access to safe water and sanitation infrastructure. FUNDING: University of California, San Diego FY19 Center Launch programme.

Extreme heat, preterm birth, and stillbirth: A global analysis across 14 lower-middle income countries.

Stillbirths and complications from preterm birth are two of the leading causes of neonatal deaths across the globe. Lower- to middle-income countries (LMICs) are experiencing some of the highest rates of these adverse birth outcomes. Research has suggested that environmental determinants, such as extreme heat, can increase the risk of preterm birth and stillbirth. Under climate change, extreme heat events have become more severe and frequent and are occurring in differential seasonal patterns. Little is known about how extreme heat affects the risk of preterm birth and stillbirth in LMICs. Thus, it is imperative to examine how exposure to extreme heat affects adverse birth outcomes in regions with some of the highest rates of preterm and stillbirths. Most of the evidence linking extreme heat and adverse birth outcomes has been generated from high-income countries (HICs) notably because measuring temperature in LMICs has proven challenging due to the scarcity of ground monitors. The paucity of health data has been an additional obstacle to study this relationship in LMICs. In this study, globally gridded meteorological data was linked with spatially and temporally resolved Demographic and Health Surveys (DHS) data on adverse birth outcomes. A global analysis of 14 LMICs was conducted per a pooled time-stratified case-crossover design with distributed-lag nonlinear models to ascertain the relationship between acute exposure to extreme heat and PTB and stillbirths. We notably found that experiencing higher maximum temperatures and smaller diurnal temperature range during the last week before birth increased the risk of preterm birth and stillbirth. This study is the first global assessment of extreme heat events and adverse birth outcomes and builds the evidence base for LMICs.

Darkness affects differentially the expression of plastid-encoded genes and delays the senescence-induced down-regulation of chloroplast transcription in cotyledons of Cucurbita pepo L. (Zucchini).

In contrast to differentiated leaves, the regulatory mechanisms of chloroplast gene expression in darkened cotyledons have not been elucidated. Although some results have been reported indicating accelerated senescence in Arabidopsis upon reillumination, the capacity of cotyledons to recover after dark stress remains unclear. We analysed the effect of two-days dark stress, applied locally or at the whole-plant level, on plastid gene expression in zucchini cotyledons. Our results showed that in the dark the overall chloroplast transcription rate was much more inhibited than the nuclear run-on transcription. While the activities of the plastid-encoded RNA polymerase (PEP) and nuclear RNA polymerase II were strongly reduced, the activities of the nuclear-encoded plastid RNA polymerase (NEP) and nuclear RNA polymerase I were less affected. During recovery upon reillumination, chloroplast transcription in the cotyledons was strongly stimulated (3-fold) compared with the naturally senescing controls, suggesting delayed senescence. Northern blot and dot blot analyses of the expression of key chloroplast-encoded photosynthetic genes showed that in contrast to psbA, which remained almost unaffected, both the transcription rate and mRNA content of psaB and rbcL were substantially decreased.

Monsoon weather and early childhood health in India.

BACKGROUND: India is expected to experience an increase in the frequency and intensity of extreme weather events in the coming decades, which poses serious risks to human health and wellbeing in the country. OBJECTIVE: This paper aims to shed light on the possible detrimental effects of monsoon weather shocks on childhood undernutrition in India using the Demographic and Health Survey 2015-16, in combination with geo-referenced climate data. METHODS: Undernutrition is captured through measures of height-for-age, weight-for-height, stunting and wasting among children aged 0-59 months. The standardised precipitation and evapotranspiration index (SPEI) is used to measure climatic conditions during critical periods of child development. RESULTS: The results of a multivariate logistic regression model show that climate anomalies experienced in utero and during infancy are associated with an increased risk of child undernutrition; exposure to excessive monsoon precipitation during these early periods of life elevates the risk of stunting, particularly for children in the tropical wet and humid sub-tropical regions. In contrast, the risk of stunting is reduced for children residing in the mountainous areas who have experienced excessive monsoon precipitation during infancy. The evidence on the short-term effects of climate shocks on wasting is inconclusive. We additionally show that excessive precipitation, particularly during the monsoon season, is associated with an increased risk of contracting diarrhoea among children under five. Diseases transmitted through water, such as diarrhoea, could be one important channel through which excessive rainfall increases the risk of stunting. CONCLUSIONS: We find a positive association between childhood undernutrition and exposure to excessive monsoon precipitation in India. Pronounced differences across climate zones are found. The findings of the present analysis warn of the urgent need to provide health assistance to children in flood-prone areas.

Electrogravimetry and Structural Properties of Thin Silicon Layers Deposited in Sulfolane and Ionic Liquid Electrolytes.

Potentiostatic deposition of silicon is performed in sulfolane (SL) and ionic liquid (IL) electrolytes. Electrochemical quartz crystal microbalance with damping monitoring (EQCM-D) is used as main analytical tool for the characterization of the reduction process. The apparent molar mass (Mapp) is applied for in situ estimation of the layer contamination. By means of this approach, appropriate electrolyte composition and substrate type are selected to optimize the structural properties of the layers. The application of SL electrolyte results in silicon deposition with higher efficiency compared to the IL 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)imide, [BMP][TFSI]. This has been associated with the instability of the IL in the presence of silicon tetrachloride and the enhanced incorporation of IL decomposition products into the growing silicon deposit. X-ray photoelectron spectroscopy (XPS) analysis supports the results about the layer composition, as suggested from the microgravimetric experiments. Attention has been given to the impact of practically relevant substrates (i.e., Cu, Ni, and vitreous carbon) on the reduction process. An effective deposition can be carried out on the metal electrodes in both electrolytes due to accelerated reaction kinetics for these types of substrates. However, on vitreous carbon (VC), a successful reduction of SiCl4 can only be accomplished in the IL, while the electroreduction process in SL is dominated by the decomposition of the electrolyte. For short deposition times, the scanning electron microscopy (SEM) images display rough morphologies in the nanometer range, which evolve further to structures with increased length scale of the surface roughness. The development of a rough interface during deposition, resulting in QCM damping at advanced stages of the process, is interpreted by a model accounting for the resistive force caused by the interaction of the liquid with a nonuniform layer interface. By using this approach, the individual contributions of the surface roughness and viscoelastic effects to the measured damping values are estimated.

The Hordeum bulbosum 25S-18S rDNA region: comparison with Hordeum vulgare and other Triticeae.

Hordeum vulgare and Hordeum bulbosum are two closely related barley species, which share a common H genome. H. vulgare has two nucleolar organizer regions (NORs), while the NOR of H. bulbosum is only one. We sequenced the 2.5 kb 25S-18S region in the rDNA of H. bulbosum and compared it to the same region in H. vulgare as well as to the other Triticeae. The region includes an intergenic spacer (IGS) with a number of subrepeats, a promoter, and an external transcribed spacer (5'ETS). The IGS of H. bulbosum downstream of 25S rRNA contains two 143-bp repeats and six 128-bp repeats. In contrast, the IGS in H. vulgare contains an array of seven 79-bp repeats and a varying number of 135-bp repeats. The 135-bp repeats in H. vulgare and the 128-bp repeats in H. bulbosum show similarity. Compared to H. vulgare, the 5'ETS of H. bulbosum is shorter. Additionally, the 5'ETS regions in H. bulbosum and H. vulgare diverged faster than in other Triticeae genera. Alignment of the Triticeae promoter sequences suggests that in Hordeum, as in diploid Triticum, transcription starts with guanine and not with adenine as it is in many other plants.

EPR study on ligand-exchange reaction between bis(diethyldiselenocarbamato)copper(II) and some bis(acetylacetonato)copper(II) complexes.

The paper is a study on the formation and properties of mixed-chelate copper(II) complexes, in which one ligand is strongly covalently bound diselenocarbamate (dsc), and the other belongs to a series of differently substituted acetylacetonates (acac), all forming weak covalent bonds. Thenoyltrifluoro- and trifluoro- substituted acetylacetonates only partly form mixed-chelate complexes, stable in toluene, benzene or dichloromethane, but gradually decomposing in chloroform or carbontetrachloride by analogy with Cu(Et2dsc)2 behavior in these solvents. Hexafluoro-acetylacetonato copper(II) completely turns into a mixed-chelate Cu(dsc)(hfacac), which remains unchanged for more than 8 months of monitoring in all solvents used. The stability of Cu(dsc)(hfacac) is attributed to the reduced remaining negative charge on selenium atom embarrassing the formation of weak D-A complex with haloalkanes. The obtained EPR parameters suggest significant lowering of the contribution of the 4s AO of copper(II) in the mixed-chelate complexes. It is shown that Cu(acac)2 does not react with Cu(Et2dsc)2.

EPR study on the ligand-exchange reaction between bis(diethyldiselenocarbamato)copper(II) and bis(octyldithiocarbonato)copper(II).

EPR study on the ligand-exchange reaction between bis(diethyldiselenocarbamato)copper(II), Cu(Et2dsc)2, and bis(octyldithiocarbonato)copper(II), Cu(octxant)2, in CH2Cl2, CHCl3, CCl4, C6H6 and C6H5.CH3 is reported for the first time. Mixing of equimolar amounts of the parents (chromophores CuSe4 and CuS4, respectively) in C6H6, C6H5.CH3 and CH2Cl2 makes EPR signals of both parents superimposed by the spectrum of a mixed-chelate Cu(xant)(dsc) complex (chromophore CuS2Se2). A new additional EPR spectrum appears in CHCl3 or CCl4 due to a five-coordinate mixed-ligand complex with the chromophore Cu(S3Se)S as follows by comparing the g-values of parents and mixed-ligand complexes. The appearance of this complex could be explained having in mind donor-acceptor properties of complexes, solvents and the resultant reaction of Cu(octxant)2 with the ester of diselenocarbamic acid yielded in Cu(Et2dsc)2 destruction by CCl4 or CHCl3.

In situ, Reversible Gating of a Mechanosensitive Ion Channel through Protein-Lipid Interactions.

Understanding the functioning of ion channels, as well as utilizing their properties for biochemical applications requires control over channel activity. Herein we report a reversible control over the functioning of a mechanosensitive ion channel by interfering with its interaction with the lipid bilayer. The mechanosensitive channel of large conductance from Escherichia coli is reconstituted into liposomes and activated to its different sub-open states by titrating lysophosphatidylcholine (LPC) into the lipid bilayer. Activated channels are closed back by the removal of LPC out of the membrane by bovine serum albumin (BSA). Electron paramagnetic resonance spectra showed the LPC-dose-dependent gradual opening of the channel pore in the form of incrementally increasing spin label mobility and decreasing spin-spin interaction. A method to reversibly open and close mechanosensitive channels to distinct sub-open conformations during their journey from the closed to the fully open state enables detailed structural studies to follow the conformational changes during channel functioning. The ability of BSA to revert the action of LPC opens new perspectives for the functional studies of other membrane proteins that are known to be activated by LPC.

Mapping of unmethylated sites in rDNA repeats in barley NOR deletion line.

Extensive cytosine methylation is characteristic of plant rDNA. Evidence exists, however, that the active rRNA genes are less methylated. In this work we report on the mapping of unmethylated CCGG sites in Hordeum vulgare rDNA repeats by digestion with methylation sensitive restriction enzyme HpaII and indirect end-labeling of the generated fragments. For mapping we used genomic DNA from barley deletion line with a single NOR on chromosome 5H. This NOR is more active in order to compensate for the missing NOR 6H. The enhanced NOR 5H activity in the deletion mutant is not due to higher multiplicity of the rRNA genes or, as sequencing showed, to changes in the subunit structure of the intergenic spacer. The HpaII sites in barley rDNA are heavily methylated. Nevertheless, a fraction of the rDNA repeats is hypomethylated with unmethylated CCGG sites at various positions. One unmethylated CCGG sequence is close to the transcription start site, downstream of the 135bp subrepeats. Unmethylated sites are also present in the external transcribed spacer and in the genes coding mature rRNAs. The patterns of unmethylated sites in the barley deletion line and in lines with two NORs were compared. It is hypothesized that the occurrence of unmethylated sites on a fixed subset of rDNA repeats correlates with their transcriptional activity.