Difficulties in achieving and maintaining the goal of measles elimination in Bulgaria.

Measles is a disease with almost a hundred year history of existing registration in Bulgaria and has been subject to mass immunization since 1969. In 2017, after a three-year period with a low number of measles cases registered, an epidemic upsurge has been recorded affecting 3 of the total 28 regions in the country. The purpose of this study was to analyze the changes of measles morbidity in Bulgaria over the period of 1921-2017 and focus on the epidemiological characteristics of the last outbreak in the region of Plovdiv in 2017. For the whole period (1921-2017) the average measles morbidity amounts to 157.69%ооо, decreasing from 525.02%ооо in 1921 to 2.32%ооо in 2017, with 99.5% rate of reduction. In the period prior to vaccination the average morbidity was 247.14%ооо while in the vaccination period it was 70.08%ооо. The ARIMA model could be used as a short-term forecast to predict the morbidity rate. Against the background of the downward tendency in morbidity this study reports a small measles outbreak in Plovdiv, involving 141 cases, after three years of no local virus transmission in the country. An unvaccinated child who returned from abroad probably imported the disease. The measles cases were mainly Roma children and a large number of them were infants. The low intensity and prolong course of the outbreak was indicative of relatively high vaccination coverage of the population. To achieve measles elimination goals, efforts must be made to strengthen surveillance and increase the vaccination coverage, targeting children and especially Roma children.

Insights into the morphological pattern of erythrocytes' aging: Coupling quantitative AFM data to microcalorimetry and Raman spectroscopy.

Erythrocytes (RBCs) constitute a very interesting class of cells both for their physiological function and for a variety of peculiarities. Due to their exceptionally strong relationship with the environment, the morphology and nanoscale characteristics of these cells can reveal their biochemical status and structural integrity. Among the possible subjects of investigations, the RBCs' ageing is of the utmost importance. This is a fundamental phenomenon that, in physiological conditions, triggers the cell turnover and ensures the blood homeostasis. With these premises, in recent years, we have presented an atomic force microscopy-based methodology to characterize the patterns of RBC ageing from the morphological point of view. In the present work, we used an ageing protocol more similar to the physiological conditions and we used differential scanning calorimetry and atomic force microscopy to probe the cross correlation between important structural and functional proteins. We also assessed the role played by fundamental structural and membrane proteins in the development of the most relevant morphological intermediates observed along the ageing. Furthermore, we coupled the morphological ageing patterns to the (bio)chemical alterations detected by Raman spectroscopy. This allowed identifying the chronology of the ageing morphologies and the metabolic pathways most involved in their development. As a whole, the present study provides the base to correlate specific molecular alterations to the development of structural anomalies, and these latter to the functional status of blood cells.

Heat- and light-induced detachment of the light-harvesting antenna complexes of photosystem I in isolated stroma thylakoid membranes.

The multisubunit pigment-protein complex of photosystem I (PSI) consists of a core and peripheral light-harvesting antenna (LHCI). PSI is thought to be a rather rigid system and very little is known about its structural and functional flexibility. Recent data, however, suggest LHCI detachment from the PSI supercomplex upon heat and light treatments. Furthermore, it was suggested that the splitting off of LHCI acts as a safety valve for PSI core upon photoinhibition (Alboresi et al., 2009). In this work we analyzed the heat- and light-induced reorganizations in isolated PSI vesicles (stroma membrane vesicles enriched in PSI). Using differential scanning calorimetry we revealed a stepwise disassembly of PSI supercomplex above 50°C. Circular dichroism, sucrose gradient centrifugation and 77K fluorescence experiments identified the sequence of events of PSI destabilization: 3min heating at 60°C or 40min white light illumination at 25°C resulted in pronounced Lhca1/4 detachment from the PSI supercomplex, which is then followed by the degradation of Lhca2/3. The similarity of the main structural effects due to heat and light treatments supports the notion that thermo-optic mechanism, structural changes induced by ultrafast local thermal transients, which has earlier been shown to be responsible for structural changes in the antenna system of photosystem II, can also regulate the assembly and functioning of PSI antenna.

Brassinosteroids regulate the thylakoid membrane architecture and the photosystem II function.

Brassinosteroids (BRs) are plant steroid hormones known to positively affect photosynthesis. In this work we investigated the architecture and function of photosynthetic membranes in mature Arabidopsis rosettes of BR gain-of-function (overexpressing the BR receptor BR INSENSITIVE 1 (BRI1), BRI1OE) and loss-of-function (bri1-116 with inactive BRI1 receptor, and constitutive photomorphogenesis and dwarfism (cpd) deficient in BR biosynthesis) mutants. Data from atomic force microscopy, circular dichroism, fluorescence spectroscopy and polarographic determination of oxygen yields revealed major structural (enlarged thylakoids, smaller photosystem II supercomplexes) and functional (strongly inhibited oxygen evolution, reduced photosystem II quantum yield) changes in all the mutants with altered BR response compared to the wild type plants. The recorded thermal dependences showed severe thermal instability of the oxygen yields in the BR mutant plants. Our results suggest that an optimal BR level is required for the normal thylakoid structure and function.

Monitoring photosynthesis in individual cells of Synechocystis sp. PCC 6803 on a picosecond timescale.

Picosecond fluorescence kinetics of wild-type (WT) and mutant cells of Synechocystis sp. PCC 6803, were studied at the ensemble level with a streak-camera and at the cell level using fluorescence-lifetime-imaging microscopy (FLIM). The FLIM measurements are in good agreement with the ensemble measurements, but they (can) unveil variations between and within cells. The BE mutant cells, devoid of photosystem II (PSII) and of the light-harvesting phycobilisomes, allowed the study of photosystem I (PSI) in vivo for the first time, and the observed 6-ps equilibration process and 25-ps trapping process are the same as found previously for isolated PSI. No major differences are detected between different cells. The PAL mutant cells, devoid of phycobilisomes, show four lifetimes: ∼20 ps (PSI and PSII), ∼80 ps, ∼440 ps, and 2.8 ns (all due to PSII), but not all cells are identical and variations in the kinetics are traced back to differences in the PSI/PSII ratio. Finally, FLIM measurements on WT cells reveal that in some cells or parts of cells, phycobilisomes are disconnected from PSI/PSII. It is argued that the FLIM setup used can become instrumental in unraveling photosynthetic regulation mechanisms in the future.