Intracellular delivery of protein nanoparticles (NP) is required for nanomedicine. Our research was focused on the quantitative analysis of protein NP intracellular accumulation and biodegradation in ...dynamics along with host cytokine gene expression. Fluorescent NP fabricated by nanoprecipitation without cross-linking of bovine serum albumin (BSA) and human immunoglobulins (hIgG) pre-labeled with Rhodamine B were non-toxic for human cells. Similar gradual uptake of the NP during 2 days and subsequent slowdown until background values for 5 days for human cell lines and donor blood mononuclear cells revealed that NP internalization was neither cell-type nor protein-specific. NP delivery into cells was inhibited by homologous and heterologous NP but did not depend on the presence of BSA or hIgG in culture media. The protein NP internalization induced interferon α, β, λ but neither γ nor interleukin 4 and 6 gene expression. Accordingly, cellular uptake of non-toxic protein NP induced Th1 polarized innate response.
Cellular uptake and biodegradation of albumin and immunoglobulin nanoparticles but not original proteins induce Th1 innate immunity. Display omitted
Moose (Alces alces) recombinant chymosin with a milk-clotting activity of 86 AU/mL was synthesized in the Kluyveromyces lactis expression system. After precipitation with ammonium sulfate and ...chromatographic purification, a sample of genetically engineered moose chymosin with a specific milk-clotting activity of 15,768 AU/mg was obtained, which was used for extensive biochemical characterization of the enzyme. The threshold of the thermal stability of moose chymosin was 55 °C; its complete inactivation occurred after heating at 60 °C. The total proteolytic activity of moose chymosin was 0.332 A280 units. The ratio of milk-clotting and total proteolytic activities of the enzyme was 0.8. The Km, kcat and kcat/Km values of moose chymosin were 4.7 μM, 98.7 s−1, and 21.1 μM−1 s−1, respectively. The pattern of change in the coagulation activity as a function of pH and Ca2+ concentration was consistent with the requirements for milk coagulants for cheese making. The optimum temperature of the enzyme was 50–55 °C. The introduction of Mg2+, Zn2+, Co2+, Ba2+, Fe2+, Mn2+, Ca2+, and Cu2+ into milk activated the coagulation ability of moose chymosin, while Ni ions on the contrary inhibited its activity. Using previously published data, we compared the biochemical properties of recombinant moose chymosin produced in bacterial (Escherichia coli) and yeast (K. lactis) producers.
. is polyploid grass species that grows in the eastern part of the Fertile Crescent, Afghanistan, and Middle Asia. It consists of tetraploid (4x) and hexaploid (6x) cytotypes (2
= 4x = 28, D
D ...(Abdolmalaki et al., 2019) X
X
and 2n = 6x = 42, D
D (Abdolmalaki et al., 2019) X
X
D
D (Adams and Wendel, 2005), respectively) that are similar morphologically. Although many
species were used in wheat breeding, the genetic potential of
.
has not yet been exploited due to its uncertain origin and significant genome modifications. Tetraploid
.
is thought to be the oldest polyploid
species, the subgenomes of which still retain some features of its ancient diploid progenitors. The D
and D
subgenomes of
.
were contributed by
(2
= 2x = 14, DD), while the X
subgenome donor is still unknown. Owing to its ancient origin,
.
can serve as model for studying genome evolution. Despite this,
.
is poorly studied genetically and no genome sequences were available for this species. We performed low-coverage genome sequencing of 4x and 6x cytotypes of
.
, and four
.
accessions belonging to different subspecies; diploid wheatgrass
(J
genome), which is phylogenetically close to D (sub)genome species, was taken as an outgroup. Subsequent data analysis using the pipeline RepeatExplorer2 allowed us to characterize the repeatomes of these species and identify several satellite sequences. Some of these sequences are novel, while others are found to be homologous to already known satellite sequences of Triticeae species. The copy number of satellite repeats in genomes of different species and their subgenome (D
or X
) affinity in
.
were assessed by means of comparative bioinformatic analysis combined with quantitative PCR (qPCR). Fluorescence
hybridization (FISH) was performed to map newly identified satellite repeats on chromosomes of common wheat,
, 4x and 6x
.
,
.
, and
.
. The new FISH markers can be used in phylogenetic analyses of the Triticeae for chromosome identification and the assessment of their subgenome affinities and for evaluation of genome/chromosome constitution of wide hybrids or polyploid species.
Background The liver fluke, Opisthorchis felineus, is widely distributed throughout Europe and large parts of the Russian Federation. In Western Siberia, information about opisthorchiasis is lacking ...although infection may lead to severe liver and bile duct diseases. We aimed to assess the current prevalence of O. felineus infection along with associated risk factors and morbidity in rural Western Siberia. Methods We conducted a community-based, cross-sectional study in the rural Shegarskiy district, Tomsk Oblast, Russian Federation. All household members (greater than or equal to 7 years) present on the survey day were enrolled (n = 600). Two stool samples per person were examined for helminth eggs, using PARASEP (DiaSys Ltd, UK). The number of eggs per gram (EPG) of feces was recorded. Each study participant was interviewed to determine risk factors, using a pre-tested questionnaire. An abdominal ultrasonography examination of liver and bile ducts was performed with a mobile, high resolution ultrasound device. In total, 488 persons completed assessments (two stool samples, completed questionnaires); of those, 436 individuals had an ultrasonography (US) examination. Results We observed a prevalence of O. felineus infection of 60.2%. Significant risk factors for infection were the consumption of river fish (odds ratio from adjusted analysis aOR 2.4, 95% CI 1.52-3.95, p0.001), particularly stock fish (OR from multivariable analysis mOR 3.2, 95% CI 2.63-3.80, p0.001), smoked fish (mOR 1.5, 95% CI 1.24-1.72, p0.001), frozen fish (mOR 1.6, 95% CI 1.29-2.02, p0.001), and raw fish (mOR 1.4, 95% CI 1.05-1.84, p = 0.02); and fishing activities (mOR 1.2, 95% CI 1.03-1.43, p = 0.019). Women had a higher risk of infection than men. Infection was associated positively with age and negatively with socio-economic status. The respondents general awareness of opisthorchiasis was quite high (93.2%), but their knowledge about infection transmission and prevention was insufficient. Children aged 7-18 years old had a lower level of awareness compared to adults. The abdominal ultrasonography results demonstrated a strong association between O. felineus infection and gallbladder stones (mOR 2.8, 95% CI 1.33-6.04, p = 0.007) and periductal fibrosis of intrahepatic bile ducts (mOR 1.9, 95% CI 1.08-3.46, p = 0.026). Conclusion O. felineus infection is highly prevalent in rural regions of Western Siberia, and associated with severe hepatobiliary pathology. Identified risk factors will be used to develop a comprehensive targeted O. felineus infection control program.
Due to the ability of influenza virus to develop drug resistance, the search for novel antivirals is an important goal of medical science and health care systems. We assessed the ability of the ...influenza virus to develop resistance to the hemagglutinin inhibitor camphecene and characterized laboratory-selected resistant strains. We showed by electron microscopy that camphecene decreases the number of virions fusing their envelopes with endosomal membranes. A 160-fold decrease in virus susceptibility was observed after six passages in cells. This was associated with the emergence of a V458L mutation in the HA2 subunit of HA and with a decrease in viral pathogenicity. Molecular modeling predicts that this substitution results in a more stable HA molecule compared to wild-type HA; and an altered camphecene-binding site. Therefore, despite the relatively rapid development of resistance, camphecene remains promising as a potential antiviral due to the low pathogenicity of resistant viruses that may arise.
Immobilized bacterial cells are presently widely used in the development of bacterial preparations for the bioremediation of contaminated environmental objects. Oil hydrocarbons are among the most ...abundant pollutants. We have previously described a new biocomposite material containing hydrocarbon-oxidizing bacteria (HOB) embedded in silanol-humate gels (SHG) based on humates and aminopropyltriethoxysilane (APTES); high viable cell titer was maintained in this material for at least 12 months. The goal of the work was to describe the ways of long-term HOB survival in SHG and the relevant morphotypes using the techniques of microbiology, instrumental analytical chemistry and biochemistry, and electron microscopy. Bacteria surviving in SHG were characterized by: (1) capacity for rapid reactivation (growth and hydrocarbon oxidation) in fresh medium; (2) ability to synthesize surface-active compounds, which was not observed in the cultures stored without SHG); (3) elevated stress resistance (ability to grow at high Cu
and NaCl concentrations); (4) physiological heterogeneity of the populations, which contained the stationary hypometabolic cells, cystlike anabiotic dormant forms (DF), and ultrasmall cells; (5) occurrence of piles in many cells, which were probably used to exchange genetic material; (6) modification of the phase variants spectrum in the population growing after long-term storage in SHG; and (7) oxidation of ethanol and acetate by HOB populations stored in SHG. The combination of the physiological and cytomorphological properties of the cells surviving in SHG for long periods may indicate a new type of long-term bacterial survival, i.e., in a hypometabolic state.
qPCR as a Selective Tool for Cytogenetics Divashuk, Mikhail G; Nikitina, Ekaterina A; Sokolova, Victoria M ...
Plants,
12/2022, Letnik:
12, Številka:
1
Journal Article
Recenzirano
Odprti dostop
qPCR is widely used in quantitative studies of plant genomes and transcriptomes. In this article, this method is considered as an auxiliary step in the preparation and selection of markers for FISH ...analysis. Several cases from the authors' research on populations of the same species were reviewed, and a comparison of the closely related species, as well as the adaptation of the markers, based on satellite tandem repeats (TRs) using quantitative qPCR data was conducted. In the selected cases, TRs with contrast abundance were identified in the cases of the
and
species, and the transfer of TRs between the wheat and related species was demonstrated. TRs with intraspecific copy number variation were revealed in
and wheat-wheatgrass partial amphidiploids, and the TR showing predominant hybridization to the sea buckthorn Y chromosome was identified. Additionally, problems such as the absence of a reference gene for qPCR, and low-efficiency and self-complementary primers, were illustrated. In the cases considered here, the qPCR results clearly show high correlation with the subsequent results of the FISH analysis, which confirms the value of this method for cytogenetic studies.
Late blight (LB) caused by the oomycete
(Mont.) de Bary is the greatest threat to potato production worldwide. Current potato breeding for LB resistance heavily depends on the introduction of new ...genes for resistance to
(
genes). Such genes have been discovered in highly diverse wild, primitive, and cultivated species of tuber-bearing potatoes (
L. section
Dumort.) and introgressed into the elite potato cultivars by hybridization and transgenic complementation. Unfortunately, even the most resistant potato varieties have been overcome by LB due to the arrival of new pathogen strains and their rapid evolution. Therefore, novel sources for germplasm enhancement comprising the broad-spectrum
genes are in high demand with breeders who aim to provide durable LB resistance. The Genbank of the N.I. Vavilov Institute of Plant Genetic Resources (VIR) in St. Petersburg harbors one of the world's largest collections of potato and potato relatives. In this study, LB resistance was evaluated in a core selection representing 20 species of seven
series according to the Hawkes (1990) classification:
(Rydb.) Hawkes,
Buk.,
Buk.,
Bitt.,
(Rydb.) Hawkes,
(Rydb.) Hawkes (wild and cultivated species), and
Corr. LB resistance was assessed in 96 accessions representing 18 species in the laboratory test with detached leaves using a highly virulent and aggressive isolate of
. The
species notably differed in their LB resistance:
Dun.,
Lindl.,
Lindl., and
Hawkes stood out at a high frequency of resistant accessions (7-9 points on a 9-point scale). Well-established specific SCAR markers of ten
genes-
,
,
,
,
,
,
and
-were used to mine 117 accessions representing 20 species from seven
series. In particular, our evidence confirmed the diverse
gene location in two American continents. The structural homologs of the
,
,
and
genes were found in the North American species other than
, the species that was the original source of these genes for early potato breeding, and in some cases, in the South American
species. The
orthologs from
and
Schlechtd et Bché were restricted to genome B in the Mesoamerican series
,
and
. The structural homologs of the
gene that were initially identified in the South American species
Hawkes and Hjert. were reported, for the first time, in the North American series of
species.
The prevalence of the Opisthorchis felineus infection in Western Siberia in 1980–2000 (review of Russian literature).
Display omitted
•Review of Russian literature on O. felineus infection prevalence ...was performed.•Western Siberia is the biggest endemic region of opisthorchiasis in Russia.•The highest prevalence of infection is detected in Tyumenskaya Oblast – above 60%.•The prevalence of infection is low in children and increases with age.•Development of control program is needed to decrease infection distribution.
In this study we reviewed Russian scientific literature (scientific publications, book chapters, monographs) published between 1 January 1979 and 31 August 2015 from two sources: Main database of the Russian Scientific Electronic Library (eLIBRARY, http://elibrary.ru/), and the Scientific Medical Library of Siberian State Medical University (http://medlib.tomsk.ru/). Specifically, the review details the infection prevalence of Opisthorchis felineus (O. felineus) in Western Siberia, Russian Federation. From the primary key words screening, 1591 records were identified from which 32 Russian-language publications were relevant. The lowest O. felineus infection rate of 0.4% was reported in Tatarstan Republic, and the highest reached 83.9% in the Khanty-Mansiysk Autonomous Okrug. The infection prevalence was lower in children than in adults and increased with age.
O. felineus infection was detected more often in indigenous population than in migrants. Infection intensity in western regions (Permskaya, Bryanskaya Oblast) was low and varied from 15 to 336 eggs per gram stool (epg), while in endemic regions it reached more than 2000 epg. In some settlements the mean intensity infection was 5234 epg. The high rates of intensity were registered in regions with a high prevalence of infection. Based on obtained data, a map of O. felineus infection prevalence in Western Siberia was developed. After mapping the results, the highest prevalence was detected in Tyumenskaya Oblast with over 60%, while the Tomskaya Oblast had the lowest prevalence at fewer than 19.0%. Khanty-Mansiysk Autonomus Okrug, Altaiskii Krai, Novosibirskaya Oblast and Omskaya Oblast had an average level of O. felineus infection of 20–39%. According to the results of the review, Western Siberia must be considered as highly endemic region for opisthorchiasis in the Russian Federation. The development of a control program specific for the Russian community is warranted.
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