Lipid Inclusions Research Articles

Exposure to different cobalt chloride levels produces oxidative stress and lipidomic changes and affects the liver structure of Cyprinus carpio juveniles.

The present investigation was undertaken to evaluate the toxic effects of CoCl2-induced hepatotoxicity and fatty acid changes in juvenile Cyprinus carpio. Fish were divided into six experimental groups in duplicate. The first group served as controls. The second group received the lowest exposure dose at 2.5µg/L. In the third group, fish were exposed to 25µg/L of CoCl2. The fourth group was exposed to 50µg/L of CoCl2. The last two groups were exposed to the highest doses, 100 and 500µg/L of CoCl2. Total antioxidant activities were estimated using a colorimetric method. Liver fatty acid compositions were analyzed by high-performance gas chromatography (GC). Hepatopathy was identified through microscopic analysis. Exposure of C. carpio to CoCl2 resulted in hepatotoxicity, indicated by increased levels of malondialdehyde (MDA), hydrogen peroxide (H2O2), protein carbonyls (PCO), and alterations in the ferric reducing antioxidant power system (FRAP). Superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), reduced glutathione (GSH), metallothioneins (MTs), and low thiol levels (L-SH) significantly increased, particularly under exposure to the highest CoCl2 doses (100 and 500µg/L). Acetylcholinesterase activity decreased significantly in C. carpio exposed to graded CoCl2 doses. Additionally, there was a decrease in polyunsaturated fatty acids (PUFA), primarily n-3 PUFA, docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), while an increase in monounsaturated (MUFA) and saturated fatty acids (SFA), including palmitic (C16:0), stearic (C18:0), palmitoleic (C16:1), and oleic (C18:1) acids, was observed. Histopathological examination of the liver confirmed hepatopathy revealing characteristic tissue changes such as leucocyte infiltration, hepatic cell membrane degradation, vacuolization, and lipid inclusions. The study provided ethnophysiology insights into the responses of C. carpio to CoCl2-induced oxidative stress and lipidomic alteration, underscoring its potential as a bioindicator for assessing environmental impacts and metal contamination.

Fine structure of the gonadal tissue of the horned octopod Eledone cirrhosa (Lamarck, 1798) (Mollusca - Octopoda) during sexual maturity.

The horned octopod Eledone cirrhosa, a medium-sized species found in Arabian Gulf off Saudi Arabia, was collected monthly from the Arabian Gulf off Dammam city during January to December 2022. Samples were dissected and prepared for examination using transmission electron microscopy. During genital maturation, seminiferous tubules are established in the testis, with active spermatogonia dividing. Spermatocytes 1 are observed in the tubule, followed by an increase in spermatogonia and spermatocytes in August. Spermiogenesis begins, with spherical spermatids evolving into elongated spermatids. In September, active spermatogonia, meiotic divisions, and increased spermiogenesis continue. Spermatozoa appear in Needham's pouch, indicating sexual maturity. The ovary undergoes various stages of development, with oocytes at stage I in June and July, followed by stage II in October and November. In stage III, follicular cords invade the oocyte's cytoplasm, forming numerous lipid inclusions and protein granules. The cytoplasm contains cisternae of endoplasmic reticulum and a poorly developed Golgi apparatus. Stage IV occurs in November, characterized by the maximum development of follicular cords and the beginning of vitellogenesis. The ooplasm contains numerous lipid inclusions, a syncytium, and secretory cells. From December, stage V oocytes are mainly present, indicating the activity phase of maximum secretion. Yolk platelets accumulate in the oocyte ooplasm, and chorion forms at the zona pellucida. In January, the first smooth eggs are found in some octopuses' ovary, with their proportion increasing steadily. This study aimed to investigate the mitogenic action of gonadotropin and identify the periods of intense cell multiplication during the sexual cycle using cytological methods.

Phenotypic and metabolic adaptations of Rhodococcus cerastii strain IEGM 1243 to separate and combined effects of diclofenac and ibuprofen.

The increasing use of non-steroidal anti-inflammatory drugs (NSAIDs) has raised concerns regarding their environmental impact. To address this, understanding the effects of NSAIDs on bacteria is crucial for bioremediation efforts in pharmaceutical-contaminated environments. The primary challenge in breaking down persistent compounds lies not in the biochemical pathways but in capacity of bacteria to surmount stressors. In this study, we examined the biodegradative activity, morphological and physiological changes, and ultrastructural adaptations of Rhodococcus cerastii strain IEGM 1243 when exposed to ibuprofen, diclofenac, and their mixture. Our findings revealed that R. cerastii IEGM 1243 exhibited moderate biodegradative activity towards the tested NSAIDs. Cellular respiration assay showed higher metabolic activity in the presence of NSAIDs, indicating their influence on bacterial metabolism. Furthermore, catalase activity in R. cerastii IEGM 1243 exposed to NSAIDs showed an initial decrease followed by fluctuations, with the most significant changes observed in the presence of DCF and the NSAID mixture, likely influenced by bacterial growth phases, active NSAID degradation, and the formation of multicellular aggregates, suggesting potential intercellular synergy and task distribution within the bacterial community. Morphometric analysis demonstrated alterations in size, shape, and surface roughness of cells exposed to NSAIDs, with a decrease in surface area and volume, and an increase in surface area-to-volume ratio (SA/V). Moreover, for the first time, transmission electron microscopy confirmed the presence of lipid inclusions, polyphosphates, and intracellular membrane-like structures in the ibuprofen-treated cells. These results provide valuable insights into the adaptive responses of R. cerastii IEGM 1243 to NSAIDs, shedding light on the possible interaction between bacteria and pharmaceutical compounds in the environment.

The relationship between ApoE4 and lipid dysregulation in glia of the PS19 mouse model of tauopathy

AbstractBackgroundApolipoprotein E4 (apoE), the major lipoprotein in the brain, plays an essential role in brain`s lipid homeostasis; furthermore, the presence of the apoE4 allele is the strongest genetic risk factor for developing late‐onset AD compared to most common apoE3 and protective apoE2 alleles. Recent studies in iPSC microglia and astrocytes demonstrate that apoE modulates cholesterol dysfunction and inflammatory responses in an isoform‐specific fashion. However, it is not known if lipid accumulation in glia mediates toxic effects in vivo in the context of tauopathy and neurodegeneration.MethodIn this work we used the combination unbiased lipidomics coupled with immunostainings to dissect the role of apoE and apoE isoforms in glial lipid metabolism of tauopathy P301S tau transgenic mice with a targeted replacement of murine apoE with human ApoE4, ApoE3, or ApoE KO. We further treated the P301S/ApoE4 mice with the LXR agonist GW3965 to test if promoting cholesterol efflux in P301S mice would reduce tau pathology and neurodegeneration in this model.ResultsBy performing unbiased lipidomic analysis of forebrain samples, we demonstrate that P301S/ApoE KI mice accumulate significant amounts of specific cholesteryl esters, oxidized sterols and endolysosomal BMP lipids in an ApoE‐isoform and Tau‐dependent manner. We further show that microglia from aged 9.5 months old P301S/ApoE4 mice accumulate significant amounts of neutral BODIPY‐positive lipid inclusions within lysosomes. Next, we demonstrate that the oral administration of the LXR agonist GW3965 for 3 months significantly reduces p‐tau‐positive AT8 immunostaining in cortex and hippocampus, protects from associated brain neurodegeneration, and improves nesting behavior in 9.5 months old P301S/ApoE4 mice. Using bulk RNA sequencing and immunostaining, we show that LXR agonist diet induces changes in cholesterol biosynthesis and metabolism that result in a significant reduction of neuroinflammatory responses in aged P301S/ApoE4 mice. Lastly, using unbiased lipidomics of forebrain tissues we demonstrate that P301S/ApoE4 mice on LXR diet exhibit significantly lower levels of cholesterol esters when compared to tau transgenic animals on control diet.ConclusionsTogether, we demonstrate that ApoE4 promotes lipid accumulation in glia in the setting of tauopathy and that reducing the accumulation of these lipids by LXR activation is neuroprotective.

Alteration of Average Thickness of Lipid Bilayer by Membrane-Deforming Inclusions.

Thickness of lipid bilayer membranes is a key physical parameter determining membrane permeability and stability with respect to formation of through pores. Most membrane inclusions or impurities like amphipathic peptides, transmembrane peptides, lipid inclusions of a different molecular shape, lipid domains, and protein-lipid domains, locally deform the membrane. The detailed structure of the locally deformed region of the membrane is a kind of "fingerprint" for the inclusion type. However, most experimental methods allow determining only averaged parameters of membranes with incorporated inclusions, thus preventing the direct obtaining of the characteristics of the inclusion. Here we developed a model that allows the obtaining of characteristic parameters of three types of membrane inclusions (amphipathic peptides, transmembrane peptides, monolayer lipid patches) from experimentally observable dependencies of the average thickness of lipid bilayer on the surface concentration of the inclusions. In the case of amphipathic peptides, the model provided the peptide parameters that were in qualitative agreement with the available experimental data.

Intrabacterial lipid inclusion-associated proteins: a core machinery conserved from saprophyte Actinobacteria to the human pathogen Mycobacterium tuberculosis.

Mycobacterium tuberculosis (Mtb), the aetiologic agent of tuberculosis (TB), stores triacylglycerol (TAG) in the form of intrabacterial lipid inclusions (ILI) to survive and chronically persist within its host. These highly energetic molecules represent a major source of carbon to support bacterial persistence and reactivation, thus playing a leading role in TB pathogenesis. However, despite its physiological and clinical relevance, ILI metabolism in Mtb remains poorly understood. Recent discoveries have suggested that several ILI-associated proteins might be widely conserved across TAG-producing prokaryotes, but still very little is known regarding the nature and the biological functions of these proteins. Herein, we performed an in silico analysis of three independent ILI-associated proteomes previously reported to computationally define a potential core ILI-associated proteome, referred to as ILIome. Our investigation revealed the presence of 70 orthologous proteins that were strictly conserved, thereby defining a minimal ILIome core. We further narrowed our analysis to proteins involved in lipid metabolism and discuss here their putative biological functions, along with their molecular interactions and dynamics at the surface of these bacterial organelles. We also highlight the experimental limitations of the original proteomic investigations and of the present bioinformatic analysis, while describing new technological approaches and presenting biological perspectives in the field. The in silico investigation presented here aims at providing useful datasets that could constitute a scientific resource of broad interest for the mycobacterial community, with the ultimate goal of enlightening ILI metabolism in prokaryotes with a special emphasis on Mtb pathogenesis.

Characteristics of a New Halotolerant Arctic Strain of Carotenogenic Microalga <i>Halochlorella rubescens</i> NAMSU SBB-20

Green microalgae capable of accumulating secondary carotenoids are the most important objects of biotechnology, and the search for new strains with unique properties, in particular, those adapted to growth at low temperatures and high salinity in the environment, is an urgent task. The NAMSU SBB-20 microalga strain was isolated from an algal-bacterial biofilm found on the coast of the White Sea in the littoral zone of the Solovetsky Archipelago. Identification of the strain showed its belonging to the species Halochlorella rubescens P.J.L.Dangeard. The species H. rubescens was first described for the White Sea. Under conditions of high light intensity, ultrastructural changes in cells are shown, among which destruction of the photosynthetic apparatus and the formation of cytoplasmic and chloroplast lipid inclusions are noted. It was shown that the culture of the NAMSU SBB-20 strain is capable of acquiring an orange color under unfavorable growth conditions. An assessment was made of the effect of the composition of the medium and the intensity of illumination on the pigment composition of the algae. The highest absolute values of the accumulation of carotenoids were noted during cultivation in light with an intensity of 150 mol PAR quanta/m2/s on BG-11 media containing no source of phosphorus (15.66 0.18 mg/L) or nitrogen (15.95 0.56 mg/L). The described strain has a biotechnological potential due to the initial halotolerance and the accumulation of high values of secondary carotenoids in the biomass.

Experimental envenomation with honeybee venom melittin and phospholipase A2 induced multiple ultrastructural changes in adrenocortical mitochondria

Bee stings represent a public health subject, but the mechanisms involved in bee venom toxicity are not yet fully understood. To evaluate the reactions of adrenocortical cells, through which organisms respond to stress, two honeybee venom components: melittin (Mlt) and phospholipase A2 (PLA2) were tested as potential chemical stressors. Modifications were investigated with transmission electron microscopy and microanalysis. A single dose of Mlt (31 mg/kg) or PLA2 (9.3 mg/kg) was injected in rats of groups ML and PL; daily doses of Mlt (350 μg/kg) or PLA2 (105 μg/kg) were injected 30 days in rats of groups M30 and P30. Adrenocortical cells in ML group showed ultrastructural degenerative alterations of nuclei, endoplasmic reticulum, and mitochondria that exhibited lipid inclusions and mitochondrial cristae (MC) re-organized into mono- or multimembrane large vesicles, and whorls of membranes. Many MC were degenerated. In the M30 group, similar ultrastructural changes, but of lower amplitude were noted; lipid cytosolic droplets were heterogenous. MC diameters in Mlt groups (melittin treated groups) were significantly higher than in control (C) group. In PL group, mitochondria contained large lipid inclusions, vesicular MC of different sizes and multiple membranes, and debris, or whorl structures. In P30 group MC were tubular with increased diameters. In both PLA2 groups (PLA2 treated groups) MC were significantly larger than in C group. We concluded that Mlt and PLA2 were powerful stressors, toxic at the tested doses, cellular reactions concerning in all groups mainly mitochondria, but also other cellular compartments. Apart from degenerative regression of MC, the rearrangement of tubular MC occurred into one or multiple large multimembrane vesicular MC. Reactions to the high doses were more pronounced, with the highest amplitude in ML group, and the lowest in P30 group.

Raman Spectroscopic Imaging of Human Bladder Resectates towards Intraoperative Cancer Assessment.

Raman spectroscopy offers label-free assessment of bladder tissue for in vivo and ex vivo intraoperative applications. In a retrospective study, control and cancer specimens were prepared from ten human bladder resectates. Raman microspectroscopic images were collected from whole tissue samples in a closed chamber at 785 nm laser excitation using a 20× objective lens and 250 µm step size. Without further preprocessing, Raman images were decomposed by the hyperspectral unmixing algorithm vertex component analysis into endmember spectra and their abundancies. Hierarchical cluster analysis distinguished endmember Raman spectra that were assigned to normal bladder, bladder cancer, necrosis, epithelium and lipid inclusions. Interestingly, Raman spectra of microplastic particles, pigments or carotenoids were detected in 13 out of 20 specimens inside tissue and near tissue margins and their identity was confirmed by spectral library surveys. Hypotheses about the origin of these foreign materials are discussed. In conclusion, our Raman workflow and data processing protocol with minimal user interference offers advantages for future clinical translation such as intraoperative tumor detection and label-free material identification in complex matrices.

Comparative survey of mitochondrial ultrastructure in IDH1-mutant astrocytoma and IDH1-wildtype glioblastoma (GBM)

ABSTRACT Gliomas are the most common malignant brain tumors with poor prognosis. The WHO’s classification recognizes isocitrate dehydrogenase 1 (IDH1) mutant astrocytoma and IDH1-wildtype glioblastoma (GBM). The IDH1 mutation confers a survival advantage over the wildtype. There are several explanations for the metabolic advantage of the IDH1 mutation, some involve mitochondrial implications. Since an ultrastructural comparison of both tumor genotypes is still lacking, we surveyed the ultrastructural effects of the IDH1 mutation on the mitochondria of the IDH1-mutant astrocytoma (n = 15) and IDH1-wildtype glioblastoma (n = 15) tumors. Our results show that both IDH1 genotypes have degenerate and uncoupled mitochondria; this has not been reported before. The presence of ample lipid inclusions and lipid droplets in the cytoplasm of both genotypes support our conclusion of dysfunctional uncoupled mitochondria. Thus, the IDH1 mutation may have no ultrastructural consequences on the mitochondria, and the aberrant mitochondria in both genotypes may be the result of other unknown mutations. The status of the mitochondria in these genotypes portends a clinical challenge since tumor cells with uncoupled mitochondria are more primitive, aggressive, and considerably treatment resistant.

Влияние введения мексидола на фоне 60-дневного воздействия тартразина на ультраструктуру эндокриноцитов коркового вещества надпочечных желёз крыс

Methods. In the experiment, 30 white mature rats were divided into 5 groups: Group 1 — control, Group 2 and Group 3 — rats exposed to tartrazine at the rate of 750 and 1500 mg/kg for 60 days. In the Group 4 and Group 5, mexidol was injected intramuscularly at the rate of 50 mg/kg against the background of the conditions of the Group 2 and Group 3. Electron microscopic examination of the adrenal glands was performed according to the standard protocol. Results. In the Group 2, epithelial cells of the glomerular and fascicular zones have a round or oval nucleus with uneven contours. The nucleus contains accumulations of heterochromatin under the karyolemma and in the karyoplasm. Large mitochondria with vesicular cristae are observed in the cytoplasm. The cisterns of the smooth endoplasmic reticulum are poorly developed. Lipid inclusions are located in groups or singly. In the Group 3, mitochondria with partially destroyed cristae were detected in the cytoplasm of endocrinocytes of the zona glomerulosa, and in the epithelial cells of the zona fasciculata, loosely packed lamellar structures were detected in the center or periphery of mitochondria, as well as cells with apoptosis. Endocrinocytes of the zona reticularis have a small size, an oval nucleus with slightly tortuous contours and accumulations of heterochromatin in the karyoplasm in the rats of the Group 2. Mitochondria were large and medium in size with spherical cristae in cross section, the cisterns of the smooth endoplasmic reticulum are unevenly expanded and fill the gaps between mitochondria. In the Group 3, almost half of the large mitochondria have densely packed lamellar structures along the periphery or in the center. Conclusion. The above-established ultramicroscopic features of endocrinocytes of the adrenal cortex after 60 days of exposure to tartrazine indicate a decrease in their secretory activity, which depends on the dose of the supplement administered. The introduction of mexidol against this background helps to restore the disturbed ultramicroscopic picture of cells to control values, which is more pronounced in the Group 4.

A historical look using virtual microscopy: the first case report of adrenomyeloneuropathy (AMN).

The history of adrenoleukodystrophy (ALD), adrenomyeloneuropathy (AMN) and other peroxisomal diseases is exemplary for the stunning progress of scientific medicine within the past 50 years. Like many breakthroughs in medicine, the detailed analysis of patients' pathologically affected tissues was instrumental, resulting in stepwise systematic clarification of what had remained enigmatic until the 1970s. This flashback paper is a recollection of the first neuropathological description of a slowly evolving clinical phenotype, spastic paraparesis with adrenal insufficiency, in a young adult by Budka et al. 1976 [3], using virtual microscopy of the original histologic slides. The clinico-pathological presentation derives from the classical cerebral ALD phenotype in boys, where electron microscopy demonstrated the underlying pathological hallmark of characteristic lipid inclusions shared by both phenotypes. Our report allowed the delineation of a new disease type almost simultaneously described in more cases as AMN by Griffin et al. 1977 [4] and Schaumburg et al. 1977 [11]. Moreover, our report indicated clinical heterogeneity in the ALD disease group that, as shown later, extends further to females, to Addison-only, and even to asymptomatic subjects. The gene underlying ALD was discovered in 1993 as a defect in the ABCD1 gene. Yet, it has hitherto remained unclear how the gene defect causes the strikingly broad and unpredictable phenotypic spectrum of ALD/AMN.

An Ascovirus Utilizes Different Types of Host Larval Regulated Cell Death Mechanisms To Produce and Release Vesicles.

Ascoviruses are insect-specific viruses that are thought to utilize the cellular apoptotic processes of host larvae to produce numerous virion-containing vesicles. In this study, we monitored the in vivo infection processes of Heliothis virescens ascovirus 3h (HvAV-3h) to illustrate the regulated cell death (RCD) of host cells. Transmission electron microscopic observations did not reveal any morphological markers of apoptosis in the fat bodies or hemocytes of HvAV-3h-infected Helicoverpa armigera or Spodoptera exigua larvae. However, several hemocytes showed the morphological criteria for necrosis and/or pyroptosis. Further in vitro biochemical tests were performed to confirm the RCD type of host cells after infection with HvAV-3h. Different morphological characteristics were found between the early (prior to 24 hours post-infection, [hpi]) and later (48 to 120 hpi) stages in both HvAV-3h infected larval fat bodies and hemocytes. In the early stages, the virions could only be found in several adipohemocytes, and the fat bodies were cleaving their contained lipid inclusions into small lipid dots. In the later stage, both fat bodies and hemocytes were filled with numerous virions. According to the morphological characteristics of HvAV-3h infected larval fat bodies or hemocytes, the pathogenic characteristics and infection patterns of HvAV-3h in the host larvae were described, and the systematic pathogenic mode of ascovirus infection was refined in this study. This study details the complete infection process of ascoviruses, which provides insights into the relationship between a pathogenesis of an insect virus and the RCD of different host tissues at different stages of infection. IMPORTANCE Viruses and other pathogens can interrupt host cellular apoptosis to gain benefits, such as sufficient resources and a stable environment that enables them to complete their replication and assembly. It is unusual for viruses to code proteins with homology to caspases, which are commonly recognized as apoptosis regulators. Ascoviruses are insect viruses with special cytopathology, and they have been hypothesized to induce apoptosis in their host larvae via coding a caspase-like protein. This enables them to utilize the process of cellular apoptosis to facilitate vesicle formation and replication. However, our previous studies revealed different trends. The fat bodies and hemocytes of Heliothis virescens ascovirus 3h (HvAV-3h)-infected larvae did not show any morphological markers of apoptosis but did display necrosis and/or pyroptosis morphological characteristics. The pathogenic characteristics and infection patterns of HvAV-3h in the host larvae were described, which can help us understand the relationship between the pathogenesis of an insect virus and host RCD.

The High Plasticity of Nonpathogenic Mycobacterium brumae Induces Rapid Changes in Its Lipid Profile during Pellicle Maturation: The Potential of This Bacterium as a Versatile Cell Factory for Lipid Compounds of Therapeutic Interest.

The immunomodulatory potential of mycobacteria to be used for therapeutic purposes varies by species and culture conditions and is closely related to mycobacterial lipid composition. Although the lipids present in the mycobacterial cell wall are relevant, lipids are mainly stored in intracellular lipid inclusions (ILIs), which have emerged as a crucial structure in understanding mycobacteria-host interaction. Little is known about ILI ultrastructure, production, and composition in nonpathogenic species. In this study, we compared the lipid profiles of the nonpathogenic immunomodulatory agent Mycobacterium brumae during pellicle maturation under different culture conditions with qualitative and quantitative approaches by using high-resolution imaging and biochemical and composition analyses to understand ILI dynamics. The results showed wax esters, mainly in early stages of development, and acylglycerols in mature ILI composition, revealing changes in dynamics, amount, and morphometry, depending on pellicle maturation and the culture media used. Low-glycerol cultures induced ILIs with lower molecular weights which were smaller in size in comparison with the ILIs produced in glycerol-enriched media. The data also indicate the simple metabolic plasticity of lipid synthesis in M. brumae, as well as its high versatility in generating different lipid profiles. These findings provide an interesting way to enhance the production of key lipid structures via the simple modulation of cell culture conditions.

Health effects of peptides obtained from hydrolysed chicken by-products by the action of Bromelia pinguin and B. karatas proteases in Wistar rats induced with metabolic syndrome

Metabolic syndrome (MS) is considered a major public health problem because it is associated with the development of cardiovascular disease and type 2 diabetes. Bioactive peptides can play an important role in the prevention and treatment of MS. The possible health effects of peptides obtained from hydrolysed chicken by-products (CH) by the action of plant proteases from Bromelia pinguin (BP), B. karatas (BK), and bromelain (BRO) were evaluated in a model of induced MS. Thirty male Wistar rats were randomised into the following groups: (1) standard diet (STD); (2) induction of MS with a hypercaloric diet (MS+CH); (3) CH-BP 200 mg CH/kg; (4) CH-BK 200 mg CH/kg; (5) CH-BRO 200 mg CH/kg; and (6) carnosine (CAR) 50 mg of carnosine/kg of body weight. The CH decreased the glucose levels (p < 0.05) and improved the lipid profile (p < 0.05) in the serum of the groups with induced MS. Liver lesions were attenuated with a decrease in hepatic enzymatic activities (p < 0.05), and the accumulation of lipid inclusions in the liver decreased. The data showed that CH and the use of proteases to obtain peptides with health effects could be a good therapeutic alternative for individuals with MS.

Immune Status of Turkeys in Industrial Cultivation Conditions

The intensification of turkey farming is accompanied by the impact of various dangerous factors on the poultry’s body. This leads to a violation of metabolic processes, an imbalance in the absorption of nutrients, and immunity decrease in turkeys, which determines the relevance of the study in this area. In this regard, figuring out when immunity reduces and, in the future, increasing its effectiveness in the body of turkeys, improving metabolic processes, growth, and development of poultry determined the purpose of this study. It was found that during the first six weeks of intensive cultivation of turkeys, the body’s immune response, the activity of non-specific resistance factors on the influence of biological factors and the intensification of poultry feeding processes were effective. However, from 42 days of poultry rearing, depletion of the immune defence of the turkey body and a decrease in the activity of non-specific resistance factors were observed, which is manifested by a likely decrease in the content of total protein, albumins, and class G and M immunoglobulins in the blood of poultry (P<0.01). Analysis of the protein spectrum of turkey blood serum indicates an immunosuppressive state of the body, probably caused by the action of biological factors. By 42 days, the bactericidal and lysozyme activity of blood serum, phagocytic activity of neutrophils, and T-cell activity of lymphocytes decrease in turkeys. Nonspecific changes in the components of the functional element of the liver were detected, which are accompanied by an increase in the number of lipid inclusions of various sizes, destructive changes in mitochondria, and a decrease in the synthetic activity of cells. Studies of the immune state of the body of turkeys, the activity of factors of non-specific resistance and liver in the cycle of intensive cultivation from 7 to 120 days revealed the presence of a critical period of decrease in the effectiveness of the body’s protection and further adequate correction. The results obtained during this study will be used for educational purposes, implemented in growing turkeys in the production conditions of “Indychka” LLC (Sumy Oblast). They are offered to all farms in Ukraine to produce turkey farming products

Implications of drug-induced phenotypical resistance: Is isoniazid radicalizing M. tuberculosis?

BackgroundTuberculosis treatment duration is long and does not guarantee eradication of infection. Shorter treatment regimens are a critical research objective to improve uptake and reduce the risk of relapse and bacterial resistance. The explanation for the need to continue treatment after patients are culture negative remains elusive. We have previously shown that the presence of lipid inclusions in mycobacterial cells is associated with an increase in antibiotic resistance.AimWe investigate the bactericidal effect of isoniazid and rifampicin on the expression of lipid inclusions and characterize the degree of the associated phenotypic antibiotic resistance to a range of anti-tuberculosis agents in current use.MethodsAntibiotic killing effect for both M. tuberculosis and M. komossense were investigated by both hollow fiber bioreactor (HFS) studies and static time kill curve (STKC) experiments. Following STKC cultures were stained with resazurin, Sytox green and Nile red to establish their live/dead (resazurin positive/Sytox positive) and lipid inclusion status, respectively. In addition, M. komossense was studied in the hollow fiber bioreactor model (HFS) and exposed to isoniazid (H) and rifampicin (R). The MIC of current antituberculosis agents for cells from the treated hollow fiber experiments were tested.ResultsAntibiotic killing was similar for both species. For M. komossense; isoniazid was ineffective at the established MIC (1 mg/L) in the hollow fiber bioreactor but rifampicin reduced the viable count rapidly at MIC (0.4 mg/L). When the two drugs were combined at their respective MICs the killing effect was significant and greater than separately. Cells exposed to isoniazid (1x and 9x MIC) for 168 h showed considerable numbers of recoverable viable cells when compared with a combination of 1x MIC R & H where there were no viable cells detectable. For both drugs the number of lipid body positive cells increased over time and this effect was most pronounced for isoniazid and was associated with phenotypic resistance to multiple anti-tuberculosis drugs.ConclusionOur results showed that isoniazid is a potent stimulator of lipid body accumulation, culture persistence, and phenotypic resistance to multiple anti-tuberculosis drugs. These findings emphasize the importance of understanding mechanisms of drug-drug interactions and phenotypic resistance in regimen building.

Effect of chitosan on cardio-metabolic risk factors induced by high fat diet in rats

PurposeThis study aims to scrutinize the efficacy of chitosan (CT) on cardio-lipotoxic responses elicited by a high-fat diet (HF).Design/methodology/approachThirty-six male Wistar rats were distributed across six groups (n = 6): normal diet (ND), HF, ND-5%CT, HF-1%CT, HF-3%CT and HF-5%CT, for seven weeks. Blood and cardiac tissues were processed for biochemical, immunohistochemical and histopathological analyses.FindingsIngestion of HF induced hyperlipidaemia and lipid accumulation, leading to increased body and heart weight by 70.5% (p < 0.0001) and 124% (p = 0.0021), respectively, compared to ND-groups. Cardiac damage markers (creatine kinase, lactate dehydrogenase and malondialdehyde) were higher in the HF-group compared to control rats. Also, atherogenic and coronary risk indices were significantly elevated by 155% (p = 0.0044) and 174% (p = 0.0008), respectively, compared to control rats. Rats fed HF had significantly reduced cardiac antioxidants (superoxide dismutase, catalase and glutathione peroxidase) and elevated expression of NF-κB-p65 and p53 (p < 0.0001) in the cardiac tissues. Histology revealed lipid inclusions in the cardiac tissues of HF-groups. CT (1%–5%) prevented hyperlipidaemia, lipid accumulation, oxidative stress and cardiac damage in HF-fed rats, while greatly improving the histology of the cardiac tissues in HF-fed rats in a dose-dependent manner.Originality/valueTo the best of the authors’ knowledge, this is the first report of the effects of CT against cardio-lipotoxicity elicited by HF diet ingestion. The findings suggest that CT may present a safe therapeutic alternative for managing complications arising from cardio-lipotoxicity.

Activity of alkaloids from Aspidosperma nitidum against Leishmania (Leishmania) amazonensis

This study evaluated the morphological changes caused by fractions and subfractions, obtained from barks of Aspidosperna nitidum, against L. (L.) amazonensis promastigotes. The ethanolic extract (EE) obtained through the maceration of trunk barks was subjected to an acid–base partition, resulting the neutral (FN) and the alkaloid (FA) fractions, and fractionation under reflux, yielded hexane (FrHEX), dichloromethane (FrDCL), ethyl acetate (FrACoET), and methanol (FrMEOH) fractions. The FA was fractionated and three subfractions (SF5-6, SF8, and SF9) were obtained and analyzed by HPLC–DAD and 1H NMR. The antipromastigote activity of all samples was evaluated by MTT, after that, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) for the active fractions were performed. Chromatographic analyzes suggest the presence of alkaloids in EE, FN, FA, and FrDCL. The fractionation of FA led to the isolation of the indole alkaloid dihydrocorynantheol (SF8 fractions). The SF5-6, dihydrocorynantheol and SF-9 samples were active against promastigotes, while FrDCL was moderately active. The SEM analysis revealed cell rounding and changes in the flagellum of the parasites. In the TEM analysis, the treated promastigotes showed changes in flagellar pocket and kinetoplast, and presence of lipid inclusions. These results suggest that alkaloids isolated from A. nitidum are promising as leishmanicidal.

Una nueva crotamina similar de la serpiente de cascabel (Crotalus durissus cumanensis) causante de daños: estudios citotóxicos cualitativos y cuantitativos en estructuras sub-celulares y neuromusculares

A quantitative model to expose the adrenal gland sub-cellular alterations produced by crotamine-like (C-L) from rattlesnake venom during 3, 6 and 24 hours (h), and also qualitative changes on mice neuromuscular structures in vivo were observed and calculated by transmission electron microscopy. A pure crotamine-like (C-L) isoform was obtained using a cationic exchange chromatography column from the rattlesnake Crotalus durissus cumanensis venom. The C-L SDS-PAGE (15.5 %) under non-reduced conditions exhibited a molecular mass of ~3 kDa single band. The C-L in vivo qualitative experiments induced ultrastructural changes in mouse neuromuscular structures at 3, 6 and 24 h, such as reduction in the number of acetylcholine vesicles, disorganisation of the secondary synaptic clefts, enlargement of the sub-sarcolemma space and alteration of the mitochondria morphology, number and cristae. Regarding neurotoxic actions in vivo, the animals injected with C-L presented spastic paralysis of the hind limbs. The quantitative alterations studied on the capillaries, the nucleus, the mitochondria the lipid inclusions, and the smooth endoplasmic reticulum were observed from 3 to 24 h after C-L injection. As far as it is known from the literature review, there are no quantitative records of similar sub-cellular alterations caused by crotamine.