Vulnerability and resilience of the road transport industry in Poland to the COVID-19 pandemic crisis.

The research aims to examine the vulnerability and resilience of road transport enterprises in Poland to a crisis caused by the COVID-19 pandemic. In theory, we refer to the Schumpeterian perspective of creative destruction. In the empirical analysis, survey data on 500 transport companies randomly selected from the database were used. We estimated partial proportional odds models to show the factors responsible for the enterprises' vulnerability and resilience to unforeseen shock. The perspective refers to the total sample size and the division into two subgroups: micro and small and medium enterprises. To justify the results, we calculated a set of statistical indicators and tests. These models enable separating enterprises according to the vulnerability level. Transport enterprises occurred significantly vulnerable to the COVID-19 crisis, particularly the demand shock. The only factor that influenced resilience was the decrease in fuel prices, which allowed a cost reduction. The crisis showed that government aid was helpful in the short run, particularly for micro and small enterprises. The medium-sized enterprises were more resilient than micro and small ones. We formulated several recommendations to help transport enterprises to adjust in the medium term.

Spectroscopic evidence of fluorescence by 1,8-diazafluoren-9-one aggregates-A prospective new ultrasensitive method for fingerprint trace detection.

Friction ridge analysis would not have been one of the most recognized branches of forensics without molecular spectroscopy. The phenomenon of fluorescence is used on daily basis to develop latent fingerprints and to enhance those that are visible. The idea behind the research was to discover selected spectroscopic properties of 1,8-diazafluoren-9-one (DFO) in various environments. This fluorescent compound has been routinely used for decades to develop latent fingerprints due to its numerous advantages, but to this day, it has not been well-understood. Analysis of absorption, fluorescence, and excitation spectra of DFO in ethanol at high dye concentration allowed identification of aggregates in the excited state. A significant influence of the dye concentration on the fluorescence spectra and on the fluorescence excitation spectra was found. In particular, dye-host aggregation was found to be much stronger in a polar solvent. DFO aggregates are strongly fluorescent in ethanol, as can be seen from the steady-state emission spectra. The impact of excitation wavelength on the effect of fingerprint detection is presented. A new reaction medium has been proposed, ethanol, which is nontoxic in relation to the currently used one, methanol. The existence in this medium of DFO aggregates in the excited state, which significantly influences the identification of amino acids present in fingerprint traces, was observed, emitting in a wide spectral range (green light).

Extreme Value Theory in Application to Delivery Delays.

This paper uses the Extreme Value Theory (EVT) to model the rare events that appear as delivery delays in road transport. Transport delivery delays occur stochastically. Therefore, modeling such events should be done using appropriate tools due to the economic consequences of these extreme events. Additionally, we provide the estimates of the extremal index and the return level with the confidence interval to describe the clustering behavior of rare events in deliveries. The Generalized Extreme Value Distribution (GEV) parameters are estimated using the maximum likelihood method and the penalized maximum likelihood method for better small-sample properties. The findings demonstrate the advantages of EVT-based prediction and its readiness for application.

Effect of Pregnancy and Stage of Lactation on Energy Processes in Isolated Blood Cells of Dairy Cows.

INTRODUCTION: The transition period is the most challenging time for dairy cattle, which is characterised not only by negative energy balance but also by fatty tissue mobilisation. MATERIAL AND METHODS: The efficiency of energy pathways, ß-oxidation in WBC and glycolysis in RBC (based on deoxyglucose transmembrane transport) were estimated. Insulin in blood plasma was determined using ELISA. RESULTS: After calving and up to one month after delivery, a significant drop in blood plasma level was noticed, simultaneously with a rise in ß-oxidation from 18.93 ±3.64 to 30.32 ±5.28 pmol/min/mg protein in WBC. A strong negative correlation between these two indices (r = -0.68) was found. During the period of transition to lactation an increase in glucose cross-membrane transportation from 41.44 ±4.92 to 50.49 ±6.41 µmol/h/g Hb was observed. A strong positive correlation between glucose transportation in RBC and ß-oxidation in WBC (r = 0.71) was noticed. These data are in agreement with results of studies on dairy cows using liver slices from dairy cows in late pregnancy and different stages of lactation, in which changes in gene expression were analysed. CONCLUSION: It seems that measuring fatty acids oxidation and glycolysis using isolated blood cells may be an adequate and relatively simple method for energy state analysis to estimate the state of dairy cow metabolism and animal health.

Short communication: Streptococcus canis is able to establish a persistent udder infection in a dairy herd.

Bovine mastitis caused by Streptococcus canis is relatively rare. Consequently, many epidemiologic aspects of the infection, including factors that mediate crossing of host species barriers by the pathogen, infectiousness of the microorganism to the mammary gland, and the course of the disease within a herd, are still not elucidated. Therefore, the aim of the present study was to describe results of a 15-mo observation of subclinical Strep. canis mastitis on a dairy farm housing 76 lactating Holstein-Friesian cows. Upon 3 visits to the farm during a period between April 2013 and June 2014, Strep. canis was cultured from milk samples of 17 (22.4% of the herd), 7 (9.6%), and 8 (11.3%) cows, respectively. The isolates obtained were characterized phenotypically by means of the API Strep identification kit (bioMérieux, Marcy l'Etoile, France), as well as genetically by using random amplified polymorphic DNA and macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis. All strains displayed the same biochemical features, and the molecular methods revealed that the isolates belonged to a single clone or were very closely related. Results of the study indicate that Strep. canis is capable of causing intramammary infections of long duration, behaving in a contagious manner. Because a persistently infected cow may serve as the source of Strep. canis infection for other animals, effective control of this type of udder infection within a herd may require similar measures to those adopted in Streptococcus agalactiae eradication programs.

Expression patterns of HvCKX genes indicate their role in growth and reproductive development of barley.

Cytokinin oxidase/dehydrogenase proteins (CKX) are encoded by a multigene family of CKX genes with a varying number of members depending on species. For some of the genes, spectacular effects on grain production in selected cereals have been observed. Despite the fact that partial or full length sequences of most HvCKX genes in barley (Hordeum vulgare) have already been published, in most cases their specific biological functions have not been reported. Detailed expression patterns for five HvCKX genes in different organs/tissues of developing barley plants coupled with analysis of RNAi silent for two genes are presented to test the hypothesis that these expression profiles might indicate their function. Elevated expression for four of them - HvCKX1, HvCKX9, HvCKX4, and HvCKX11 - was found in developing kernels of wild-type plants compared to other tissues. HvCKX5 was mainly expressed in leaf tissue. Lower expression was noted for HvCKX1 in seedling roots and for HvCKX9 in leaves. The documented effect of RNAi silencing of HvCKX1 and a trend for HvCKX9 was higher plant productivity, and the trait was inherited through four generations. Higher plant yield was determined by higher numbers of seeds and spikes. Increased productivity was significantly greater in HvCKX1 silenced plants showing higher relative expression of HvCKX1 in developing kernels of wild-type plants compared to the expression of HvCKX9. Both HvCKX1 silenced T1 seedlings of cv. Golden Promise and the newly transformed breeding line STH7308 showed greater root mass, but this trait was not inherited in the next generation. Similarly HvCKX9 silenced T1 seedlings exhibited greater plant height without inheritance in the next generation. It is suggested that these effects were not inherited because of compensation by other genes co-ordinately regulating reproductive development. One line with untypically changed, inherited phenotype, which was selected from several dozen silenced lines showing stable and common phenotypes is presented.

HvCKX2 gene silencing by biolistic or Agrobacterium-mediated transformation in barley leads to different phenotypes.

BACKGROUND: CKX genes encode cytokinin dehydrogenase enzymes (CKX), which metabolize cytokinins in plants and influence developmental processes. The genes are expressed in different tissues and organs during development; however, their exact role in barley is poorly understood. It has already been proven that RNA interference (RNAi)-based silencing of HvCKX1 decreased the CKX level, especially in those organs which showed the highest expression, i.e. developing kernels and roots, leading to higher plant productivity and higher mass of the roots [1]. The same type of RNAi construct was applied to silence HvCKX2 and analyze the function of the gene. Two cultivars of barley were transformed with the same silencing and selection cassettes by two different methods: biolistic and via Agrobacterium. RESULTS: The mean Agrobacterium-mediated transformation efficiency of Golden Promise was 3.47% (±2.82). The transcript level of HvCKX2 in segregating progeny of T(1) lines was decreased to 34%. The reduction of the transcript in Agrobacterium-derived plants resulted in decreased CKX activity in the developing and developed leaves as well as in 7 DAP (days after pollination) spikes. The final phenotypic effect was increased productivity of T(0) plants and T(1) lines. Higher productivity was the result of the higher number of seeds and higher grain yield. It was also correlated with the higher 1000 grain weight, increased (by 7.5%) height of the plants and higher (from 0.5 to 2) numbers of spikes. The transformation efficiency of Golden Promise after biolistic transformation was more than twice as low compared to Agrobacterium. The transcript level in segregating progeny of T(1) lines was decreased to 24%. Otherwise, the enzyme activity found in the leaves of the lines after biolistic transformation, especially in cv. Golden Promise, was very high, exceeding the relative level of the control lines. These unbalanced ratios of the transcript level and the activity of the CKX enzyme negatively affected kernel germination or anther development and as a consequence setting the seeds. The final phenotypic effect was the decreased productivity of T(0) plants and T(1) lines obtained via the biolistic silencing of HvCKX2. CONCLUSION: The phenotypic result, which was higher productivity of silenced lines obtained via Agrobacterium, confirms the hypothesis that spatial and temporal differences in expression contributed to functional differentiation. The applicability of Agrobacterium-mediated transformation for gene silencing of developmentally regulated genes, like HvCKX2, was proven. Otherwise low productivity and disturbances in plant development of biolistic-silenced lines documented the unsuitability of the method. The possible reasons are discussed.

The RNA-mediated silencing of one of the Pin genes in allohexaploid wheat simultaneously decreases the expression of the other, and increases grain hardness.

The RNAi-mediated silencing of Pina and Pinb, the two genes responsible for the grain texture of allohexaploid wheat, was induced and analysed in two wheat cultivars, Kontesa and Torka. A characterization of the two genes in non-transgenic plants revealed that Pinb carries a point mutation, designated Pinb-D1c in both cultivars. This mutation does not influence transcript abundance or protein content. Two silencing cassettes of the hpRNA type were constructed and used for stable transformation via Agrobacterium. In total, 43 transgenic lines representing the two cultivars were obtained, transformed with the silencing cassettes for Pina or for Pinb or co-transformed with both cassettes. The relative transcript levels of the two genes in the same progeny plant were found to be similar, independent of the silencing cassette used. The reduction in the Pina and Pinb transcript levels in the segregating T(1) progeny of Kontesa and Torka transformed with one of the silencing cassettes exceeded 80%. Co-transformation with the silencing cassettes for both genes resulted in a reduction of over 91% of Pina and Pinb transcripts in some segregating T(1) progeny of Kontesa. The silencing was transmitted to the T(4) kernel generation of the T(3) lines. A significant reduction or lack of both puroindoline proteins in the silenced lines correlated with an essential increase in grain hardness. The discussion covers some new insights into the function of the Pin genes, including the simultaneous silencing of both, independent of the siRNA signal.

Silencing of the HvCKX1 gene decreases the cytokinin oxidase/dehydrogenase level in barley and leads to higher plant productivity.

Stable RNA interference-based technology was used to silence the expression of the HvCKX1 gene in barley and the TaCKX1 gene in wheat and triticale. The silencing cassettes containing the fragments of these genes in the sense and antisense orientations were cloned into the pMCG161 binary vector and used for Agrobacterium-based transformation. Out of the five cultivars representing the three studied species, transgenic plants were obtained from one barley cultivar Golden Promise, one wheat cultivar Kontesa, and one triticale cultivar Wanad. Almost 80% of 52 regenerated lines of Golden Promise exhibited significantly decreased cytokinin oxidase/dehydrogenase (CKX) enzyme activity in bulked samples of their T(1) roots. There was a positive correlation between the enzyme activity and the plant productivity, expressed as the yield, the number of seeds per plant, and the 1000 grain weight. Additionally, these traits were associated with a greater root mass. Lower CKX activity led to a higher plant yield and root weight. This higher plant productivity and altered plant architecture were maintained in a population of segregating T(1) plants. The levels of HvCKX1 transcript accumulation were measured in various tissues of Golden Promise and Scarlett non-transgenic barley plants in order to choose the most appropriate plant organs to study the expression and/or silencing of the gene in those transgenic lines. The highest levels of the HvCKX1 transcript were detected in spikes 0 days after pollination (0 DAP), 7 DAP, and 14 DAP, and in the seedling roots. The analysis of HvCKX1 gene expression and CKX enzyme activity and the evaluation of the phenotype were performed in the progeny of seven selected transgenic T(1) lines. The relative expression of HvCKX1 measured in the spikes 0 DAP and 14 DAP, respectively, ranged from 0.52+/-0.04 to 1.15+/-0.26 and from 0.47+/-0.07 to 0.89+/-0.15. The lowest relative values were obtained for the enzyme activity in the spikes at 0 DAP, which ranged from 0.15+/-0.02 to 1.05+/-0.14 per single progeny plant. Based on these three values, the coefficient of HvCKX1 silencing in the spikes was estimated. Possible mechanisms leading to higher plant productivity via the silencing of HvCKX1 and a decrease in CKX enzyme activity are discussed.

Brain tumor formation in tuberous sclerosis depends on Erk activation.

Tuberous sclerosis (TS) is an autosomal dominant disease associated with the formation of usually benign tumors or hamartomas. The disease is connected with upregulation of mammalian target of rapamycin, central regulator of protein translation, which is usually regarded to be activated by Akt kinase. Here, we show for the first time that in all four brain lesions and one angiomyolipoma from TS patients both extracellular signal-regulated kinase (Erk) and p90 ribosomal S6 kinase 1 activation as well as Erk-dependent phosphorylation of p70 ribosomal S6 kinase 1 are markedly elevated whereas Akt, participating in the classical pathway of mammalian target of rapamycin activation is not always activated. Erk activation is also present in TS-derived cell lines. Importantly, Erk inhibition leads to the decrease of proliferation potential of such lines. These results show that Erk is specifically implicated in the pathogenesis of hamartomas.

Upregulation of the WNT pathway in tuberous sclerosis-associated subependymal giant cell astrocytomas.

Tuberous sclerosis (TS), autosomal dominant disorder manifested by the formation of usually benign tumors in the brain, heart, kidneys and skin, results from an inactivating mutation in one of two tumor suppressor genes TSC1 or TSC2. Protein products of these genes, hamartin and tuberin, respectively, have been shown to participate in the mTOR pathway controlling translation of approx. 10-15% of all proteins. In the current paper, we aimed at verifying whether hamartin and tuberin may also be implicated in the control of gene transcription. Very recently it has been hypothesized that the pathway triggered by WNT, one of embryonic growth factors involved in cell differentiation and migration, could be disturbed in TS. In order to test this hypothesis we evaluated samples of four subependymal giant cell astrocytomas (SEGAs), brain tumors developing in the progress of TS. We found that beta-catenin, transcription factor and mediator of WNT pathway activity is indeed present and active in SEGAs. mRNA transcripts for c-Myc and N-Myc, proteins whose transcription is regulated by beta-catenin, were upregulated in two of four SEGAs, while cyclin D1 mRNA was significantly higher in three SEGAs. At the same time, c-Myc and N-Myc proteins were detected in the same two samples. Thus, we show for the first time that aberrant WNT signaling may contribute to the pathogenesis of TS-associated SEGAs.

Long-term outcome of early treatment of developmental hip dysplasia using an abduction splint.

Background. Treatment of congenital hip dysplasia, when implemented in the first weeks of life, gives a good outcome. Very few publications, however, have addressed the anatomical remodeling of the affected hip. In this study, we evaluate the anatomical outcome of the treatment applied. Material and methods. We examined 89 children diagnosed with congenital hip dysplasia in the first three months of life and then treated with an abduction device, for a total of 148 hips. The time of follow-up varied from 8 to 13 years. All these children were given a clinical examination according to McKay, along with Severin X-ray classification. Results. Before treatment was implemented, the patients had been diagnosed as types IIc (56 hips, 37.8%), IId (34 hips, 22.9%), III (40 hips, 27.1%), and IV (18 hips, 12.2%). In one hip final assessment showed signs of aseptic necrosis of the proximal femur. 147 hips showed total remodeling, and in the Severin X-ray classification scheme were evaluated as type I. Conclusions. Severin X-ray clasification is easy to implement and in our material corresponds with the clinical evaluation according to McKay. Early implementation of treatment with an abduction device is worthwhile and gives a satisfactory outcome.

[Osteogenesis imperfecta as an interdisciplinary medical problem]. / Wrodzona lamliwosc kosci jako interdyscyplinarny problem medyczny.

In years 2001-2003 six children (3 boys ond 3 girls) from 2.1 to 18.7 years old with osteogenesis imperfecta (OI) were treated in our clinic. Besides clinical and radiological evaluation, densitometry and biochemical analysis of collagen were performed. Biochemical analysis was performed on cultured in vitro fibroblasts from the skin biopsies. Based on this findings 1 child was graded as type I of OI, 4 children as type III and 1 as type IV. Recognition of collagen defect helps with diagnosis and makes the decision for pharmacological treatment easier. In 1 child with the dramatic type III OI therapy with pamidronian was implemented with good result. Performed surgical treatment, intramedullar stabilisation with Rush rod, proved to be useful choice in correcting axial deformations of the lower extremities and preventing from the future fractures.

Chromium-yeast supplementation of chicken broilers in an industrial farming system.

The experiment was performed on ISA broilers at big farm conditions. One day old chickens were randomly assigned to two poultry-houses with the capacity of 10600 birds each. Chromium was provided in the form of chromium enriched yeast (1.0 g Cr/kg yeast) given in doses of 0.2 mg Cr/kg diet. No significant differences in growth rate and only a slight decrease of feed conversion ratio was found in broilers receiving chromium-yeast supplement. However, these birds had a lower mortality rate comparied with the rate observed in the controls. Moreover, dietary chromium led also to an improvement of carcass composition. Organic chromium supplementation increased the weight of pectoral muscles and meat of these broilers contained less amounts of fat and cholesterol. These effects were accompanied by an insignificant increase of chromium concentration in the pectoral muscles of birds fed chromium supplemented diets, from 40 to 52 microg/kg wet tissue.