Genetic analysis reveals three novel QTLs underpinning a butterfly egg-induced hypersensitive response-like cell death in Brassica rapa.

BACKGROUND: Cabbage white butterflies (Pieris spp.) can be severe pests of Brassica crops such as Chinese cabbage, Pak choi (Brassica rapa) or cabbages (B. oleracea). Eggs of Pieris spp. can induce a hypersensitive response-like (HR-like) cell death which reduces egg survival in the wild black mustard (B. nigra). Unravelling the genetic basis of this egg-killing trait in Brassica crops could improve crop resistance to herbivory, reducing major crop losses and pesticides use. Here we investigated the genetic architecture of a HR-like cell death induced by P. brassicae eggs in B. rapa. RESULTS: A germplasm screening of 56 B. rapa accessions, representing the genetic and geographical diversity of a B. rapa core collection, showed phenotypic variation for cell death. An image-based phenotyping protocol was developed to accurately measure size of HR-like cell death and was then used to identify two accessions that consistently showed weak (R-o-18) or strong cell death response (L58). Screening of 160 RILs derived from these two accessions resulted in three novel QTLs for Pieris brassicae-induced cell death on chromosomes A02 (Pbc1), A03 (Pbc2), and A06 (Pbc3). The three QTLs Pbc1-3 contain cell surface receptors, intracellular receptors and other genes involved in plant immunity processes, such as ROS accumulation and cell death formation. Synteny analysis with A. thaliana suggested that Pbc1 and Pbc2 are novel QTLs associated with this trait, while Pbc3 also contains an ortholog of LecRK-I.1, a gene of A. thaliana previously associated with cell death induced by a P. brassicae egg extract. CONCLUSIONS: This study provides the first genomic regions associated with the Pieris egg-induced HR-like cell death in a Brassica crop species. It is a step closer towards unravelling the genetic basis of an egg-killing crop resistance trait, paving the way for breeders to further fine-map and validate candidate genes.

Insect egg-killing: a new front on the evolutionary arms-race between brassicaceous plants and pierid butterflies.

Evolutionary arms-races between plants and insect herbivores have long been proposed to generate key innovations such as plant toxins and detoxification mechanisms that can drive diversification of the interacting species. A novel front-line of plant defence is the killing of herbivorous insect eggs. We test whether an egg-killing plant trait has an evolutionary basis in such a plant-insect arms-race. Within the crucifer family (Brassicaceae), some species express a hypersensitive response (HR)-like necrosis underneath butterfly eggs (Pieridae) that leads to eggs desiccating or falling off the plant. We studied the phylogenetic distribution of this trait, its egg-killing effect on and elicitation by butterflies, by screening 31 Brassicales species, and nine Pieridae species. We show a clade-specific induction of strong, egg-killing HR-like necrosis mainly in species of the Brassiceae tribe including Brassica crops and close relatives. The necrosis is strongly elicited by pierid butterflies that are specialists of crucifers. Furthermore, HR-like necrosis is linked to PR1 defence gene expression, accumulation of reactive oxygen species and cell death, eventually leading to egg-killing. Our findings suggest that the plants' egg-killing trait is a new front on the evolutionary arms-race between Brassicaceae and pierid butterflies beyond the well-studied plant toxins that have evolved against their caterpillars.

Rapid decomposition of geological samples by ammonium bifluoride (NH4 HF2 ) for combined Hf-Nd-Sr isotope analyses.

RATIONALE: Complete decomposition of silicate rock matrices is crucial in determining their isotopic compositions, but acid dissolution in a high-pressure steel-jacketed bomb, which has been the only powerful, effective technique thus far, is time-consuming and expensive. Rock dissolution using ammonium bifluoride (ABF), as described here, is a viable alternative. METHODS: Geological reference materials (GRMs) were digested using ABF in closed Teflon beakers at temperatures of 220/230°C in a convection oven and subsequently treated with HNO3 . Hf-Sr-Nd were separated and purified using ion-exchange chemistry columns calibrated for 50-2 mg samples. The isotopic compositions of Sr-Nd were measured by Thermal Ionization Mass Spectrometry, while that of Hf by Multi-Collector Inductively Coupled Plasma Mass Spectrometry, both with normal 1011 Ω and gain calibrated 1013 Ω amplifiers. RESULTS: Total procedural blanks of our protocol are 0.5 ng for Sr, 0.2 ng for Nd and <25 pg for Hf. Test runs with GRMs, ranging in composition from basic to felsic and dissolved in ABF, yield accurate 87 Sr/86 Sr, 143 Nd/144 Nd and 176 Hf/177 Hf isotope ratios as compared with those obtained with the bomb dissolution technique. Reproducibilities were comparable, on the order of 10-20 ppm. Our technique allows combined Hf-Sr-Nd isotope analyses of low-mass (50-2 mg) samples. CONCLUSIONS: The ABF digestion is an alternative technique to high-pressure bomb dissolution in matrix decomposition for accurate and reproducible Hf-Nd-Sr isotope analyses of geological samples within a reasonable time (3-4 days), with high sample throughput and low costs in geochemistry and environmental sciences.

Biological Applications at the Cutting Edge of Cryo-Electron Microscopy.

Cryo-electron microscopy (cryo-EM) is a powerful tool for macromolecular to near-atomic resolution structure determination in the biological sciences. The specimen is maintained in a near-native environment within a thin film of vitreous ice and imaged in a transmission electron microscope. The images can then be processed by a number of computational methods to produce three-dimensional information. Recent advances in sample preparation, imaging, and data processing have led to tremendous growth in the field of cryo-EM by providing higher resolution structures and the ability to investigate macromolecules within the context of the cell. Here, we review developments in sample preparation methods and substrates, detectors, phase plates, and cryo-correlative light and electron microscopy that have contributed to this expansion. We also have included specific biological applications.

Change in the Caries Status of Pregnant Women in Southeast Hungary and Its Background.

PURPOSE: To examine the caries status of pregnant women in southeast Hungary and compare the results with a similar study completed in the same region ten years earlier to determine whether any improvement has taken place in the past decade. Another objective was to assess the effects of age, socioeconomic status and number of pregnancies on oral health. MATERIALS AND METHODS: The dental examination of 130 pregnant women was carried out at the Dental Clinic of University of Szeged, Faculty of Dentistry in 2011-2013 (group 1). Caries status was examined according to WHO guidelines. In order to compare demographic and socioeconomic factors with DMF indices, find the decisive influencing factors and compare the results with the results of the previous study (group 2), two-way ANOVA and regression analyses were conducted. RESULTS: The number of highly educated, professional, urban participants included in the present group (group 1) was greater compared with that of group 2. Despite the higher age of the pregnant women in present study, statistically significantly lower DMFT (10.05 vs 12.57, p = 0.008) and DMFS (19.41 vs 26.26, p = 0.001) scores were found than ten years ago. In both studies, the regression analyses showed that women's age was the main contributing factor to caries status. CONCLUSION: The lower DMFT and DMFS values and the participants' willingness to participate in the dental screening in the present study group were probably the result of their higher educational background and that they worked as professionals.

Evaluation of participatory planning: Lessons from Hungarian Natura 2000 management planning processes.

Stakeholder participation in nature conservation policies and especially in the management of protected areas has gained importance in the last decades. These changes are underlined by democratic principles and the perceived contribution of stakeholder involvement to the effectiveness of conservation management. Evaluating participatory processes is essential to learn about the past and thus increase the quality of future processes. The evaluation can be useful for the organisations responsible for planning and management, stakeholders and policy makers as well. The present paper shows the results of a systematic evaluation of 25 participatory processes related to the development of management plans for Natura 2000 sites in Hungary between 2007 and 2015. A conceptual framework was developed to evaluate the process and outcome of participatory management planning processes. Criteria were based on the scientific literature on public participation and tailored to conservation-related management planning and stakeholder involvement. Evaluated processes were grouped in three cases based on their time range and financial sources. Overall, the analysed processes scored at a medium level, showing better performance in the process criteria than in the outcome criteria. The best case scored significantly higher in four criteria compared to the other cases: representativeness, resource availability for facilitation, new, creative ideas and impact on the plan. The main factors behind the success were (1) embeddedness of the planning process in a larger project, where the plan was a tool for conservation, (2) carrying out only one process at a time, (3) previous experience of facilitators and planners with participatory planning and (4) the opportunity and capacity to propose a payment scheme as an incentive. But even this case received low scores in some criteria: conflict resolution, early involvement and well defined goals. Based on the results we suggest that more data is needed to evaluate the implementation of the plans and, in many cases, the impact of the process on the plan. Performance can be improved with the assistance of policy makers by further developing guidelines, harmonising payment schemes with the conservation measures of the plans and providing training on conflict resolution. The evaluation framework proved to be suitable for the assessment of a large set of conservation related management planning processes, but it also had some limitations, e.g. concerning the incorporation of stakeholders' views in the evaluation.

Structural analysis of respiratory syncytial virus reveals the position of M2-1 between the matrix protein and the ribonucleoprotein complex.

UNLABELLED: Respiratory syncytial virus (RSV), a member of the Paramyxoviridae family of nonsegmented, negative-sense, single-stranded RNA genome viruses, is a leading cause of lower respiratory tract infections in infants, young children, and the elderly or immunocompromised. There are many open questions regarding the processes that regulate human RSV (hRSV) assembly and budding. Here, using cryo-electron tomography, we identified virus particles that were spherical, filamentous, and asymmetric in structure, all within the same virus preparation. The three particle morphologies maintained a similar organization of the surface glycoproteins, matrix protein (M), M2-1, and the ribonucleoprotein (RNP). RNP filaments were traced in three dimensions (3D), and their total length was calculated. The measurements revealed the inclusion of multiple full-length genome copies per particle. RNP was associated with the membrane whenever the M layer was present. The amount of M coverage ranged from 24% to 86% in the different morphologies. Using fluorescence light microscopy (fLM), direct stochastic optical reconstruction microscopy (dSTORM), and a proximity ligation assay (PLA), we provide evidence illustrating that M2-1 is located between RNP and M in isolated viral particles. In addition, regular spacing of the M2-1 densities was resolved when hRSV viruses were imaged using Zernike phase contrast (ZPC) cryo-electron tomography. Our studies provide a more complete characterization of the hRSV virion structure and substantiation that M and M2-1 regulate virus organization. IMPORTANCE: hRSV is a leading cause of lower respiratory tract infections in infants and young children as well as elderly or immunocompromised individuals. We used cryo-electron tomography and Zernike phase contrast cryo-electron tomography to visualize populations of purified hRSV in 3D. We observed the three distinct morphologies, spherical, filamentous, and asymmetric, which maintained comparable organizational profiles. Depending on the virus morphology examined, the amount of M ranged from 24% to 86%. We complemented the cryo-imaging studies with fluorescence microscopy, dSTORM, and a proximity ligation assay to provide additional evidence that M2-1 is incorporated into viral particles and is positioned between M and RNP. The results highlight the impact of M and M2-1 on the regulation of hRSV organization.

Mosquito Attraction: Crucial Role of Carbon Dioxide in Formulation of a Five-Component Blend of Human-Derived Volatiles.

Behavioral responses of the malaria mosquito Anopheles coluzzii (An. gambiae sensu stricto molecular 'M form') to an expanded blend of human-derived volatiles were assessed in a dual-port olfactometer. A previously documented attractive three-component blend consisting of NH3, (S)-lactic acid, and tetradecanoic acid served as the basis for expansion. Adding 4.5% CO2 to the basic blend significantly enhanced its attractiveness. Expansion of the blend with four human-derived C4-volatiles was then assessed, both with and without CO2. Only when CO2 was offered simultaneously, did addition of a specific concentration of 3-methyl-1-butanol or 3-methyl-butanoic acid significantly enhance attraction. The functional group at the terminal C of the 3-methyl-substituted C4 compounds influenced behavioral effectiveness. In the absence of CO2, addition of three concentrations of butan-1-amine caused inhibition when added to the basic blend. In contrast, when CO2 was added, butan-1-amine added to the basic blend strongly enhanced attraction at all five concentrations tested, the lowest being 100,000 times diluted. The reversal of inhibition to attraction by adding CO2 is unique in the class Insecta. We subsequently augmented the three-component basic blend by adding both butan-1-amine and 3-methyl-1-butanol and optimizing their concentrations in the presence of CO2 in order to significantly enhance the attractiveness to An. coluzzii compared to the three- and four-component blends. This novel blend holds potential to enhance malaria vector control based on behavioral disruption.

A stabilized headless measles virus attachment protein stalk efficiently triggers membrane fusion.

Paramyxovirus attachment and fusion (F) envelope glycoprotein complexes mediate membrane fusion required for viral entry. The measles virus (MeV) attachment (H) protein stalk domain is thought to directly engage F for fusion promotion. However, past attempts to generate truncated, fusion-triggering-competent H-stem constructs remained fruitless. In this study, we addressed the problem by testing the hypothesis that truncated MeV H stalks may require stabilizing oligomerization tags to maintain intracellular transport competence and F-triggering activity. We engineered H-stems of different lengths with added 4-helix bundle tetramerization domains and demonstrate restored cell surface expression, efficient interaction with F, and fusion promotion activity of these constructs. The stability of the 4-helix bundle tags and the relative orientations of the helical wheels of H-stems and oligomerization tags govern the kinetics of fusion promotion, revealing a balance between H stalk conformational stability and F-triggering activity. Recombinant MeV particles expressing a bioactive H-stem construct in the place of full-length H are viable, albeit severely growth impaired. Overall, we demonstrate that the MeV H stalk represents the effector domain for MeV F triggering. Fusion promotion appears linked to the conformational flexibility of the stalk, which must be tightly regulated in viral particles to ensure efficient virus entry. While the pathways toward assembly of functional fusion complexes may differ among diverse members of the paramyxovirus family, central elements of the triggering machinery emerge as highly conserved.

An archived serum sample as a clue for identifying the primary source of a nosocomial hepatitis C virus outbreak in a haemodialysis unit.

Due to an unexpected technical error, patients at a dialysis unit who were seronegative for hepatitis C virus (HCV) were temporarily transferred to another dialysis unit next to a ward reserved for HCV-seropositive patients. In the following 7 months, 17 patients were diagnosed as anti-HCV positive. The aim of the study was to reveal the cause of this nosocomial infection. Anti-HCV-positive sera were further tested by molecular methods. Data collection and on-site epidemiologic inspections were carried out. The source of the nosocomial infection proved to be a seropositive patient treated at the unit, who died before the outbreak was recognized. The exact date of the infection was determined.

Capturing enveloped viruses on affinity grids for downstream cryo-electron microscopy applications.

Electron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.

[Postoperative monitoring of children with obstructive sleep apnea syndrome]. / Obstruktív alvási apnoe szindrómában szenvedo gyermekek posztoperatív monitorozása.

INTRODUCTION: Treatment of pediatric obstructive sleep apnea syndrome is surgical. The incidence of postoperative respiratory complications in this population is 5-25%. AIM: The aim of the authors was to present the preoperative evaluation and monitoring procedure elaborated in Heim Pál Children Hospital, Budapest. METHOD: 142 patients were involved in the study. Patient history was obtained and physical examination was performed in all cases. Thereafter, polysomnography was carried out, the severity of the obstructive sleep apnea syndrome was determined, and the patients underwent tonsilloadenotomy. RESULTS: 45 patients with mild, 50 patients with moderate and 47 patients with severe obstructive sleep apnea syndrome were diagnosed. There was no complication in patients with mild disease, while complications were observed in 6 patients in the moderate group and 24 patients in the severe group (desaturation, apnea, stridor, stop breathing) (p<0.000). In patients with severe obstructive sleep apnea syndrome, no significant difference was noted in preoperative apnoea-hypapnea index (p = 0.23) and in nadir oxygen saturation values (p = 0.73) between patients with and without complication. CONCLUSIONS: Patients with severe obstructive sleep apnea syndrome should be treated in hospital where pediatric intensive care unit is available.

Effect of active and passive techniques used in thromboembolic prophylaxis on venous flow velocity in the post-procedure period.

Introduction: Studies have shown that using mechanical thromboembolic prophylaxis methods speeds up venous flow and decreases stasis. These studies examine the post-intervention period of 1-10 min. The length of the effect of procedures to raise venous flow velocity cannot be determined by clinical trials. To apply mathematical techniques to estimate how long mechanical thromboembolism prophylaxis procedures will increase venous flow rate. Methods: In the survey, we examined 25 persons (poststroke patients), with an average age of 57.2 ± 6.3 years. Regarding the proportion of genders, 13 (52%) participants were male and 12 (48%) female. The peak venous blood flow velocity was measured with a HADECO BIDOP ES-100V II type Doppler ultrasound device, using an 8 MHz head, in the femoral vein, at the level of the hip joint. We estimated the change of the venous blood flow velocity from the available sampled data using the method of least squares. For the calculations, we used Microsoft Excel, version Mac Excel 2019. Results: The decrease in peak venous flow velocity can be approximated by a logarithm function. Mathematical calculations show that after active thromboembolic prophylaxis interventions, resting venous flow velocity is restored at 26.8 min on the intact limb and 85.1 min on the hemiparetic side. Resting flow velocity is restored in 131.9 min after passive mobilization of the hemiparetic side and in 137.7 min after the consensual effect. Discussion: An elementary mathematical function can be used to estimate the time to recovery of peak venous flow velocity to resting state from measurements taken 15 min after the intervention. Active and passive mechanical thromboembolic prophylaxis after the intervention has a longer-term effect on venous flow velocity.

Absence of Nkx2-3 induces ectopic lymphatic endothelial differentiation associated with impaired extramedullary stress hematopoiesis in the spleen.

The red and white pulps as two main parts of the spleen are arranged around distinct types of vasculature, and perform significantly different functions in both humans and mice. Previous observations indicated a profound alteration of the local vessel specialization in mice lacking Nkx2-3 homeodomain transcription factor, including contradictory results suggesting presence of an ectopic lymphatic vascular structure. Furthermore, how the absence of Nkx2-3 and the consequential changes in endothelial components affect the extramedullary hematopoietic activity restricted to the splenic red pulp is unknown. In this work, we investigated the role of Nkx2-3 homeodomain transcription factor as a major morphogenic determinant for vascular specification, and its effect in the extramedullary hematopoiesis following acute blood loss and pharmacological stimulation of megakaryocyte differentiation after treatment with thrombopoietin-receptor mimetic Romiplostim. We found that, in mice lacking Nkx2-3, Prox1-positive lymphatic capillaries containing gp38/CD31 double positive lymphatic endothelial cells develop, arranged into an extensive meshwork, while the Clever1-positive venous segments of red pulp blood vasculature are absent. This lymphatic endothelial shift is coupled with a severely compromised splenic erythropoiesis and a significantly reduced splenic megakaryocyte colony formation following Romiplostim treatment in mice lacking Nkx2-3. These findings indicate that the shift of microvascular patterning in the absence of Nkx2-3 includes the emergence of ectopic Prox1-positive lymphatic vessels, and that this pivoting towards lymph node-like vascular patterning is associated with an impaired reserve hematopoietic capacity of the splenic red pulp.

Insulin, Testosterone, and Albumin in Term and Preterm Breast Milk, Donor Milk, and Infant Formula.

BACKGROUND: Infants have three options for feeding: their own mother's breast milk, donor milk, or infant formula. Insulin, testosterone, total protein, and albumin levels were measured in breast milk samples from the first 6 months of lactation, in donor milk samples, and in different infant formulas. METHODS: Mothers who gave birth to term (n = 19) or preterm (n = 19) infants were recruited to collect breast milk samples during the first 6 months of lactation. The Breast Milk Collection Center (Unified Health Institution, Pécs, Hungary) provided 96 donor milk (DM) samples for analysis in our study. Insulin, testosterone, total protein, and albumin levels were measured in breast milk, donor milk, and infant formulas. RESULTS: During the first 2 months of lactation, the concentration of insulin was lower (-27.4%) while the testosterone concentration was higher (+20.8%) compared to the period between the 3rd and 6th months only in the preterm breast milk samples. The infant formulas examined did not contain insulin or testosterone. Holder pasteurization (HoP) did not influence the level of testosterone in human milk, although HoP decreased the insulin (-53.6%) and albumin (-38.6%) concentrations. CONCLUSIONS: Diet impacts the hormone intake of infants, underlining the importance of breastfeeding and the possible supplementation of formula-fed infants.

Validity and Reliability of the Hungarian Version of Aberdeen Varicose Vein Questionnaire.

PURPOSE: The aim of our study was to translate the Aberdeen Varicose Vein Questionnaire (AVVQ) into Hungarian, and to investigate the validity and reliability of the Hungarian AVVQ, as well as to assess the health-related quality of life in patients with varicose veins of the leg. METHODS: 374 adults participated in this study who were divided into two groups (varicose vein, healthy). We analyzed internal consistency, convergent validity (using the 36-Item Short Form Survey, SF-36), repeatability, and intra-class correlation coefficient of the Hungarian AVVQ. Regarding discriminant validity, we determined the scores of the Hungarian AVVQ in both groups using the Mann-Whitney U-test. RESULTS: The Cronbach-alpha value was 0.890, while the correlation coefficient was R = 1.000. According to the results of the convergent validation, the scores of pain and dysfunction moderately correlated with some scores of the SF-36. The score of cosmetic appearance had a relationship with many scores of the SF-36. We registered a significant relationship between the score of extent of varicosity and some scores of the SF-36. There was significant correlation between the score of complications and numerous scores of the SF-36 (physical functioning, role limitations due to physical health, pain and general health). The score of pain and dysfunction, cosmetic appearance, extent of varicosity, complications and total score of the Hungarian AVVQ showed a significant difference between both groups. CONCLUSIONS: The Hungarian AVVQ was a reliable and a valid tool to assess the health-related quality of life among patients with varicose veins and was a useful tool to justify the further treatment of the patients.

Postoperative respiratory complications in children with obstructive sleep apnoea syndrome.

Objective: We aimed to prospectively assess the effect of comorbidities on the occurrence of postoperative respiratory complications (PoRCs) after adenotonsillectomy in children with obstructive sleep apnoea syndrome (OSA) and whether otherwise healthy children need a higher level of postoperative monitoring. Methods: 577 children who had OSA and underwent adenotonsillectomy were enrolled. The effects of demographics, comorbidities and OSA on PoRCs were investigated with logistic regression analysis. Results: The PoRC rate was 4.3%. Postoperative oxygen desaturations were more marked in patients with comorbidities (p = 0.005). The presence of comorbidity increased the risk of PoRCs (odds ratio 4.234/3.226-5.241, 95% confidence intervals, p < 0.001). There was no difference in apnoea-hypopnoea index (AHI) values between comorbid patients with and without PoRCs [8.2 (3.8-50.2) vs 14.3 (11.7-23.3)]. (p = 0.37). In the group of patients without comorbidities, PoRCs were associated with a higher AHI [14.7 (3.4-51.3) vs 3.9 (2.0- 8.0), p < 0.001]. Conclusions: Comorbidities are more closely linked with PoRCs than OSA severity. In patients without comorbidity, PoRCs are associated with OSA severity and usually occur within the first 2 hours after the intervention.

Experimental Arthritis Inhibits Adult Hippocampal Neurogenesis in Mice.

Background: Adult-born neurons of the hippocampal dentate gyrus play a role in specific forms of learning, and disturbed neurogenesis seems to contribute to the development of neuropsychiatric disorders, such as major depression. Neuroinflammation inhibits adult neurogenesis, but the effect of peripheral inflammation on this form of neuroplasticity is ambiguous. Objective: Our aim was to investigate the influence of acute and chronic experimental arthritis on adult hippocampal neurogenesis and to elucidate putative regulatory mechanisms. Methods: Arthritis was triggered by subcutaneous injection of complete Freund's adjuvant (CFA) into the hind paws of adult male mice. The animals were killed either seven days (acute inflammation) or 21 days (chronic inflammation) after the CFA injection. Behavioral tests were used to demonstrate arthritis-related hypersensitivity to painful stimuli. We used in vivo bioluminescence imaging to verify local inflammation. The systemic inflammatory response was assessed by complete blood cell counts and by measurement of the cytokine/chemokine concentrations of TNF-α, IL-1α, IL-4, IL-6, IL-10, KC and MIP-2 in the inflamed hind limbs, peripheral blood and hippocampus to characterize the inflammatory responses in the periphery and in the brain. In the hippocampal dentate gyrus, the total number of newborn neurons was determined with quantitative immunohistochemistry visualizing BrdU- and doublecortin-positive cells. Microglial activation in the dentate gyrus was determined by quantifying the density of Iba1- and CD68-positive cells. Results: Both acute and chronic arthritis resulted in paw edema, mechanical and thermal hyperalgesia. We found phagocytic infiltration and increased levels of TNF-α, IL-4, IL-6, KC and MIP-2 in the inflamed hind paws. Circulating neutrophil granulocytes and IL-6 levels increased in the blood solely during the acute phase. In the dentate gyrus, chronic arthritis reduced the number of doublecortin-positive cells, and we found increased density of CD68-positive macrophages/microglia in both the acute and chronic phases. Cytokine levels, however, were not altered in the hippocampus. Conclusions: Our data suggest that acute peripheral inflammation initiates a cascade of molecular and cellular changes that eventually leads to reduced adult hippocampal neurogenesis, which was detectable only in the chronic inflammatory phase.

Thyroxine and Thyroid-Stimulating Hormone in Own Mother's Milk, Donor Milk, and Infant Formula.

Breastfeeding is widely supported in clinical and home practices, and it is known that different forms of infant feeding differ in hormone content. Thyroid hormones have essential physiological roles. In our study, we examined thyroid-stimulating hormone (TSH), thyroxine, and albumin levels in breast milk produced for term (n = 16) or preterm (n = 15) infants throughout the first 6 months of lactation. Moreover, we analyzed these components in donor human milk and in three different infant formulas. Term and preterm breast milk samples were collected monthly. The two groups had similar levels of TSH (18.4 ± 1.4 vs. 24.7 ± 2.9 nU/L), but term milk contained higher amounts of thyroxine (11,245.5 ± 73.8 vs. 671.6 ± 61.2 nmol/L) during the examination period. The albumin level was significantly higher in preterm breast milk than in term breast milk (328.6 ± 17.1 vs. 264.2 ± 6.8 mg/L). In preterm breast milk we detected downward trends in the levels of TSH (-30.2%) and thyroxine (-29.2%) in the 3rd through 6th month compared to the first 2 months of lactation. Microbiological safety of donor milk was ensured by Holder pasteurization (HoP). From the Breast Milk Collection Center of Pécs, Hungary, we enrolled 44 donor mothers into the study. HoP decreased TSH (-73.8%), thyroxine (-22.4%), and albumin (-20.9%) concentrations. Infant formulas used by the Neonatal Intensive Care Unit of the University of Pécs were found to not contain the investigated hormones, but their albumin levels were similar to the breast milk samples. The present study shows the lack of thyroid hormones in infant formulas compared to human milk and raises the question of whether formula-fed infants should be supplemented with thyroid hormones.

A structural analysis of M protein in coronavirus assembly and morphology.

The M protein of coronavirus plays a central role in virus assembly, turning cellular membranes into workshops where virus and host factors come together to make new virus particles. We investigated how M structure and organization is related to virus shape and size using cryo-electron microscopy, tomography and statistical analysis. We present evidence that suggests M can adopt two conformations and that membrane curvature is regulated by one M conformer. Elongated M protein is associated with rigidity, clusters of spikes and a relatively narrow range of membrane curvature. In contrast, compact M protein is associated with flexibility and low spike density. Analysis of several types of virus-like particles and virions revealed that S protein, N protein and genomic RNA each help to regulate virion size and variation, presumably through interactions with M. These findings provide insight into how M protein functions to promote virus assembly.