Cytoplasmic Vacuoles Research Articles

Effects of the insecticide imidacloprid on the post-embryonic development of the honey bee Apis mellifera (Hymenoptera: Apidae)

The widespread use of pesticides in agriculture has been linked to declines in bee populations worldwide. Imidacloprid is a widely used systemic insecticide that can be found in the pollen and nectar of plants and has the potential to negatively impact the development of bee larvae. We investigated the effects of oral exposure to a realistic field concentration (20.5 ng g−1) of imidacloprid on the midgut and fat body of Apis mellifera worker larvae. Our results showed that larvae exposed to imidacloprid exhibited changes in the midgut epithelium, including disorganization of the brush border, nuclear chromatin condensation, cytoplasm vacuolization, and release of cell fragments indication cell death. Additionally, histochemical analysis revealed that the midgut brush border glycocalyx was disorganized in exposed larvae. The fat body cells of imidacloprid-exposed larvae had a decrease in the size of lipid droplets from 50 to 8 μm and increase of 100 % of protein content, suggesting possible responses to the stress caused by the insecticide. However, the expression of de cdc20 gene, which plays a role in cell proliferation, was not affected in the midgut and fat body of treated larvae. These results suggest that imidacloprid negatively affects non-target organs during the larval development of A. mellifera potentially impacting this important pollinator species.

The protective effect of Nostoc commune Vauch. polysaccharide on alcohol-induced acute alcoholic liver disease and gut microbiota disturbance in mice.

In recent years, the incidence of alcoholic liver disease (ALD) has gradually increased, the development of ALD is attached great attentions. Nostoc commune Vauch. polysaccharide (NCVP) is beneficial to maintain the gut health, but the protective effect of NCVP on the liver has not been reported yet. To study the protective effect and the underlying mechanisms of NCVP on ALD, a mouse model of acute ALD was established. We built an acute ALD mouse model and explored the protective effect of NCVP through the detection of cytokines, histological examination, determination of short chain fatty acids, and 16S rRNA analysis of gut microbiota. NCVP had hepatoprotective effects on acute alcohol-induced mice by improving antioxidant capacity, reducing oxidative stress and the serum cytokine levels (IL-1β, IL-6, and TNF-α). Simultaneously, histopathological changes in liver indicated that NCVP could inhibit local hepatocyte necrosis, cytoplasmic vacuolation and inflammatory cell infiltration induced by alcohol. NCVP also increased the level of total short-chain fatty acids of acute ALD mice. In addition, NCVP could significantly decrease the Firmicutes/Bacteroidetes ratio and the abundance of Patescibacteria, Helicobacter, and Actinomycetes and increase the abundance of Lachospiraceae, Prevotellaceae-UCG-003, Lactobacillaceae, and Desulfovibrio. Our study proved that NCVP had in vivo hepatoprotective effect on acute ALD mice and provided scientific evidences that NCVP might be a promising drug candidate for the prevention and treatment of ALD.

Variations in complete blood counts and cellular morphological alterations in blood smear of COVID-19 patients from a tertiary care center

Corona viruses are a large family of viruses of which Corona virus disease 19 (COVID 19) infection evokes a hyperinflammatory response and severe acute respiratory distress syndrome in human beings. Complete blood count evaluation and peripheral smear examination is an essential tool for early diagnosis and management of COVID-19 positive patients. This study was aimed to evaluate the variations in complete blood counts (CBC) and changes in the cellular morphology in the blood smear in patients affected with COVID-19. This was an observational study, undertaken in a tertiary care center. The study included baseline blood samples from 96 proven COVID-19 positive cases. The samples were subjected for automated complete blood count analysis and peripheral smear examination.Thequantitative data were expressed by mean and Standard deviation. Qualitative categorical variables were expressed as numbers and percentages. Out of the total 96 cases, the most common quantitative finding was neutrophilic leukocytosis with relative lymphopenia. Five cases of absolute lymphopenia was observed. Majority of the cases had normal hemoglobin values and normal platelet counts. The characteristic morphological changes were seen in neutrophils and lymphocytes. The neutrophils showed presence of toxic granules, cytoplasmic vacuolations and hypolobation.The morphological alteration in lymphocytes included reactive lymphocytes with plasmacytoid and monocytoid morphology, large granular lymphocytes and Turk cells. Infrequent finding of leukoerythroblastic reaction was also observed in our study.To summarize, hematological investigation of COVID 19 cases may show several abnormal findings. Quantitative findings like Neutrophilia and lymphopenia were characteristic, whereas, the most common morphological alteration observed was atypical lymphocytes. Quantitative and qualitative alterations of hematological parameters are one of the investigations which would aid timely diagnosis of the disease. Toxic granules and cytoplasmic vacuolations in neutrophils may indicate ongoing sepsis in COVID 19 patients which in turn aids in efficient management of the patients to reduce the mortality rate.

Paraptosis: a non-classical paradigm of cell death for cancer therapy.

Due to the sustained proliferative potential of cancer cells, inducing cell death is a potential strategy for cancer therapy. Paraptosis is a mode of cell death characterized by endoplasmic reticulum (ER) and/or mitochondrial swelling and cytoplasmic vacuolization, which is less investigated. Considerable evidence shows that paraptosis can be triggered by various chemical compounds, particularly in cancer cells, thus highlighting the potential application of this non-classical mode of cell death in cancer therapy. Despite these findings, there remain significant gaps in our understanding of the role of paraptosis in cancer. In this review, we summarize the current knowledge on chemical compound-induced paraptosis. The ER and mitochondria are the two major responding organelles in chemical compound-induced paraptosis, which can be triggered by the reduction of protein degradation, disruption of sulfhydryl homeostasis, overload of mitochondrial Ca2+, and increased generation of reactive oxygen species. We also discuss the stumbling blocks to the development of this field and the direction for further research. The rational use of paraptosis might help us develop a new paradigm for cancer therapy.

Cold atmospheric plasma fumigation suppresses postharvest apple Botrytis cinerea by triggering intracellular reactive oxygen species and mitochondrial calcium

Infection by Botrytis cinerea poses a great threat to the postharvest life of apple fruit. In this study, the effects of cold atmospheric plasma (CAP) fumigation on apple B. cinerea under different exposure times and intensities were investigated. The growth of B. cinerea in vitro and in vivo was significantly suppressed by the CAP fumigation at least 700 μL/L for 5 min. To reveal the possible mechanism of antifungal activity of CAP fumigation, the pathogen was exposed to 700 μL/L and 1000 μL/L for 5 min, respectively. The results indicated that the CAP-treated spores of the pathogen underwent shrinkage, cell membrane collapse and cytoplasmic vacuolation. The results obtained from the fluorescent probe assay and flow cytometry indicated that CAP caused the accumulation of reactive oxygen species (ROS), the elevation of mitochondrial and intracellular Ca2+ levels, and the decrease in mitochondrial membrane potential of the pathogen. Investigation on statues of cell life showed that typical hallmarks of apoptosis in the CAP-treated B. cinerea spores occurred, as indicted by a large degree of increased phosphatidylserine externalization, dysfunction of membrane permeability, DNA fragmentation, distortion of morphology, chromatin condensation, and metacaspase activation observed in B. cinerea spores after CAP fumigation. Overall, CAP fumigation triggered a metacaspase-dependent apoptosis of B. cinerea spores mediated by intracellular ROS burst and Ca2+ elevation via mitochondrial dysfunction and disruption, and therefore reduced the pathogenicity of B. cinerea and suppressed postharvest Botrytis rot of apple fruit. These results would provide an insight into the underlying mechanism of CAP fumigation acting on the pathogen. The CAP fumigation makes much convenient application of CAP in storage environment to deactivate microorganism.

Clinical and laboratory factors associated with the presence of dysmorphic oocytes in intracytoplasmic sperm injection cycles.

This study investigated the clinical and laboratory factors associated with the presence of dysmorphic oocytes in intracytoplasmic sperm injection (ICSI) cycles. The study involved 200 ICSI cycles, performed from 2020 to 2021, that yielded at least one mature oocyte. Clinical characteristics and ovarian stimulation methods were compared between 68 cycles with at least one dysmorphic oocyte (the dysmorphic group) and 132 cycles with normal-form oocytes only (the non-dysmorphic group). Dysmorphic oocytes were characterized by dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body. The ages of the women, indications for in vitro fertilization, serum anti-Müllerian hormone levels, and rates of current ovarian endometrioma were similar between the dysmorphic and non-dysmorphic groups. In both groups, the three ovarian stimulation regimens, two types of pituitary suppression, and total gonadotropin dose were employed similarly. However, the dual-trigger method was used more frequently in the dysmorphic group (67.6% vs. 50%, p=0.024). The dysmorphic group contained significantly more immature oocytes and exhibited significantly lower oocyte maturity (50% vs. 66.7%, p=0.001) than the non-dysmorphic cycles. Within the dysmorphic group, significantly lower oocyte maturity was found in the cycles using a dual-trigger, but not in those with a human chorionic gonadotropin trigger. ICSI cycles with dysmorphic oocytes are closely associated with reduced oocyte maturity. This association was observed exclusively in dual-trigger cycles.

Label-Free Intracellular Multi-Specificity in Yeast Cells by Phase-Contrast Tomographic Flow Cytometry.

In-flow phase-contrast tomography provides a 3D refractive index of label-free cells in cytometry systems. Its major limitation, as with any quantitative phase imaging approach, is the lack of specificity compared to fluorescence microscopy, thus restraining its huge potentialities in single-cell analysis and diagnostics. Remarkable results in introducing specificity are obtained through artificial intelligence (AI), but only for adherent cells. However, accessing the 3D fluorescence ground truth and obtaining accurate voxel-level co-registration of image pairs for AI training is not viable for high-throughput cytometry. The recent statistical inference approach is a significant step forward for label-free specificity but remains limited to cells' nuclei. Here, a generalized computational strategy based on a self-consistent statistical inference to achieve intracellular multi-specificity is shown. Various subcellular compartments (i.e., nuclei, cytoplasmic vacuoles, the peri-vacuolar membrane area, cytoplasm, vacuole-nucleus contact site) can be identified and characterized quantitatively at different phases of the cells life cycle by using yeast cells as a biological model. Moreover, for the first time, virtual reality is introduced for handling the information content of multi-specificity in single cells. Full fruition is proofed for exploring and interacting with 3D quantitative biophysical parameters of the identified compartments on demand, thus opening the route to a metaverse for 3D microscopy.

Osimertinib induces paraptosis and TRIP13 confers resistance in glioblastoma cells

The efficacy of osimertinib, a third-generation epidermal growth factor receptor tyrosine kinase inhibitor, has been evaluated in glioblastoma (GBM) through preclinical and clinical trials. However, the underlying mechanism of osimertinib-induced GBM cell death and the underlying resistance mechanism to osimertinib remains unclear. Here, we demonstrate that Osimertinib induces paraptosis in GBM cells, as evidenced by the formation of cytoplasmic vacuoles, accumulation of ubiquitinated proteins, and upregulation of endoplasmic reticulum (ER) stress markers like CHOP. Additionally, neither apoptosis nor autophagy was involved in the osimertinib-induced cell death. RNAseq analysis revealed ER stress was the most significantly downregulated pathway upon exposure to osimertinib. Consistently, pharmacologically targeting the PERK-eIF2α axis impaired osimertinib-induced paraptosis. Notably, we show that the expression of thyroid receptor-interacting protein 13 (TRIP13), an AAA+ATPase, alleviated osimertinib-triggered paraptosis, thus conferring resistance. Intriguingly, MK-2206, an AKT inhibitor, downregulated TRIP13 levels and synergized with Osimertinib to suppress TRIP13-induced high GBM cell growth in vitro and in vivo. Together, our findings reveal a novel mechanism of action associated with the anti-GBM effects of osimertinib involving ER stress-regulated paraptosis. Furthermore, we identify a TRIP13-driven resistance mechanism against Osimertinib in GBM and offer a combination strategy using MK-2206 to overcome such resistance.

A Homozygous MAN2B1 Missense Mutation in a Doberman Pinscher Dog with Neurodegeneration, Cytoplasmic Vacuoles, Autofluorescent Storage Granules, and an α-Mannosidase Deficiency.

A 7-month-old Doberman Pinscher dog presented with progressive neurological signs and brain atrophy suggestive of a hereditary neurodegenerative disorder. The dog was euthanized due to the progression of disease signs. Microscopic examination of tissues collected at the time of euthanasia revealed massive accumulations of vacuolar inclusions in cells throughout the central nervous system, suggestive of a lysosomal storage disorder. A whole genome sequence generated with DNA from the affected dog contained a likely causal, homozygous missense variant in MAN2B1 that predicted an Asp104Gly amino acid substitution that was unique among whole genome sequences from over 4000 dogs. A lack of detectable α-mannosidase enzyme activity confirmed a diagnosis of a-mannosidosis. In addition to the vacuolar inclusions characteristic of α-mannosidosis, the dog exhibited accumulations of autofluorescent intracellular inclusions in some of the same tissues. The autofluorescence was similar to that which occurs in a group of lysosomal storage disorders called neuronal ceroid lipofuscinoses (NCLs). As in many of the NCLs, some of the storage bodies immunostained strongly for mitochondrial ATP synthase subunit c protein. This protein is not a substrate for α-mannosidase, so its accumulation and the development of storage body autofluorescence were likely due to a generalized impairment of lysosomal function secondary to the accumulation of α-mannosidase substrates. Thus, it appears that storage body autofluorescence and subunit c accumulation are not unique to the NCLs. Consistent with generalized lysosomal impairment, the affected dog exhibited accumulations of intracellular inclusions with varied and complex ultrastructural features characteristic of autophagolysosomes. Impaired autophagic flux may be a general feature of this class of disorders that contributes to disease pathology and could be a target for therapeutic intervention. In addition to storage body accumulation, glial activation indicative of neuroinflammation was observed in the brain and spinal cord of the proband.

Toxicological and morphological aspects of nano-TiO2 and nano-TiO2-Ag acute action on the liver of mice

Aim. To study the toxic effect of TiO2 and TiO2-Ag nanopowders on the morphology and elemental composition of the laboratory mice liver. Materials and methods. The study used a model of acute intoxication on laboratory animals. Mice were injected intraperitoneally with TiO2 and TiO2-Ag nanopowders at 4000 mg/kg, 7000 mg/kg, or 10000 mg/kg doses. During two weeks, the animals were observed, lethality was assessed, the accumulation of nanopowder in the organ and the morphology of liver tissues were investigated. The content of titanium and silver in liver samples was determined by optical emission spectroscopy with inductively coupled plasma. Liver tissue micropreparations were examined using an Olympus BX51 light microscope. Also, the micropreparations of the liver were examined by scanning electron microscopy (SEM) using the Tescan Mira 3 device, and the elemental composition was determined using an energy dispersive spectrometer Oxford instrument, X-max 80 mm2. Results. The dependence of the mice lethality on the nanopowders dose was revealed; mortality was higher when exposed to nano-TiO2-Ag compared to nano-TiO2. Average lethal doses were calculated using probit analysis. For nano-TiO2, the LD50 is 4783.30 mg/kg; for nano-TiO2-Ag – 724.44 mg/kg. The accumulation of titanium, titanium, and silver in the liver after exposure to nano-TiO2 and nano-TiO2-Ag was established. In general, there was a tendency to increase the content of titanium in the skin tissue with an increase in the administered dose of nanopowders. Morphological changes in the liver were studied by histological methods. The most characteristic morphological signs of the toxic effect of nano-TiO2 on tissue were dystrophic changes at the level of 67.7 % (cytoplasmic vacuolization in hepatocytes), and when exposed to nano-TiO2-Ag – initial necrotic changes at the level of 70 % (hepatocytes with nuclear pyknosis). It is worth noting that the toxic effect of nano-TiO2 and nano-TiO2-Ag is much less often manifested by focal necrosis and inflammatory reactions (focal infiltration), in some cases, there were adaptive changes that provoked an increase in the number of binuclear hepatocytes. In case of detection agglomerates of a foreign object (crystalline inclusions) were obtained, which were examined spectrally and showed a high content of titanium (Ti). SEM morphometry showed that the size of nanoparticles and their agglomerates ranged from 80 nm to 20 μm. Conclusions. The lethality of mice was higher when the composition of nano-TiO2-Ag was introduced compared to nano-TiO2. Based on the calculated average lethal doses, both nanopowders were assigned to the 3rd class (moderately dangerous) of the danger of chemical substances according to the classification of GOST 12.1.007-76. It was established that with an increase in the injected dose in the tissue of the products of laboratory mice, the accumulation of titanium (under the action of nano-TiO2) and titanium and silver (under the action of nano-TiO2-Ag) increases. Characteristic microscopic signs of the toxic effect of TiO2 and TiO2-Ag nanopowders after intraperitoneal injection in laboratory bags are dystrophic changes in hepatocytes, necrosis of parenchymal disease, while inflammatory reactions occur less often. SEM and the method of elemental mapping of titanium confirmed the presence of TiO2 nanoparticles and their agglomerates in skin tissue when TiO2 nanopowder was administered.

Characteristics of structural and functional alterations following traumatic brain injury in neurons and glial cells of the sensorimotor cortex

Introduction and Aim: Traumatic brain injury (TBI) is regarded as a significant worldwide health issue and a significant contributor to mortality and disability. The objective of this study was to examine the parameters and nature of the regenerative and destructive processes that occur in the rat cerebral cortex depending on the degree and time of the TBI. Materials and Methods: The experiments were carried out on 24 sexless adult mice weighing 180–220 g. The primary group of mice had severe TBI. To assess the severity of the TBI model, histological data, animal survival rates, and motor and cognitive dysfunctions were examined. Both light and electron microscopy were used to study the animal brains in each group. Results: The areas of injury were filled with blood, and microscopic examination revealed that the foci of contusion had destroyed brain tissue in the form of tissue and blood vessel fragments. Most dystrophic neuronal changes in mice with severe TBI between 1 and 21 days after the injury were acute neuronal swelling, hydropic dystrophy of nerve cells with clear cytoplasmic vacuolization, localized and complete chromatolysis, hyperchromatism, and homogeneous cytoplasm. Conclusion: Brain tissue lesions develop in the early stages of a TBI through rapid necrotic cell death.

Hyperhomocysteinaemia Promotes Doxorubicin-Induced Cardiotoxicity in Mice.

Doxorubicin, a widely used chemotherapeutic drug in clinical oncology, causes a series of cardiac side effects referred to as doxorubicin-induced cardiotoxicity. Hyperhomocysteinaemia is an independent risk factor for multiple cardiovascular diseases. However, whether hyperhomocysteinaemia contributes to doxorubicin-induced cardiotoxicity is currently unknown. In this study, we explored the pathogenic effects of hyperhomocysteinaemia induced by dietary methionine supplementation (2% wt/wt in rodent chow) in a mouse model of doxorubicin-induced cardiotoxicity. Our data showed that methionine supplementation doubled serum homocysteine levels, inducing mild hyperhomocysteinaemia. Doxorubicin at a cumulative dosage of 25 mg/kg body weight led to significant weight loss and severe cardiac dysfunction, which were further exacerbated by methionine-induced mild hyperhomocysteinaemia. Doxorubicin-induced cardiac atrophy, cytoplasmic vacuolisation, myofibrillar disarray and loss, as well as cardiac fibrosis, were also exacerbated by methionine-induced mild hyperhomocysteinaemia. Additional folic acid supplementation (0.006% wt/wt) prevented methionine-induced hyperhomocysteinaemia and inhibited hyperhomocysteinaemia-aggravated cardiac dysfunction and cardiomyopathy. In particular, hyperhomocysteinaemia increased both serum and cardiac oxidative stress, which could all be inhibited by folic acid supplementation. Therefore, we demonstrated for the first time that hyperhomocysteinaemia could exacerbate doxorubicin-induced cardiotoxicity in mice, and the pathogenic effects of hyperhomocysteinaemia might at least partially correlate with increased oxidative stress and could be prevented by folic acid supplementation. Our study provides preliminary experimental evidence for the assessment of hyperhomocysteinaemia as a potential risk factor for chemotherapy-induced cardiotoxicity in cancer patients.

A mixture of glyphosate and 2,4-D herbicides enhances the deleterious reproductive outcomes induced by Western diet in obese male mice.

The consumption of Western diet (WD) - enriched in fats and sugars - is associated with overweight, obesity and male reproductive disorders. In addition to WD intake, crops and dairy products display residues of herbicides, including glyphosate and 2,4-D that are widely applied worldwide. The concomitant exposure to WD and herbicides - mimicking contemporary scenarios - is not fully investigated. Thus, we evaluated the effects of glyphosate and 2,4-D, alone or in mixture, on WD-induced alterations in the male genital system. Male C57BL6J mice were submitted to WD (chow containing 20% lard, 0.2% cholesterol, 20% sucrose, and high sugar solution with 23.1 and 18.9 g/L of D-fructose and D-glucose) for 6 months. Concomitantly to WD, the animals received glyphosate (0.05, 5, or 50 mg/kg/day), 2,4-D (0.02, 2 or 20 mg/kg/day) or their mixture (0, 05 + 0.02, 5 + 2, or 50 + 20 mg/kg/day) by intragastrical administration (5×/week). Doses were based on Acceptable Daily Intake (ADIs) or No Observed Adverse Effect Level (NOAEL) values. Herbicide exposure did not alter the WD-induced obesity, hypercholesterolemia and hyperglycemia. WD induced sperm cell abnormalities, reduced the number, volume and area of Leydig cells, enhanced the frequency of epididymal abnormalities, decreased the proliferation in both germinal and epididymal epithelia, and reduced the number of androgen receptor (AR) positive epididymal cells. Remarkably, the herbicide mixtures promoted such WD-induced effects: increased the frequency of sperm cell and epididymal abnormalities (absence of sperm, cytoplasmic vacuoles, and clear cell hypertrophy) (5 + 2 and 50 + 20 doses); decreased Leydig cell nuclei volume and area (5 + 2 and 50 + 20 doses), reduced epididymal cell proliferation (all mixtures), and AR expression (50 + 20 dose). In addition, herbicide mixtures reduced serum testosterone levels (5 + 2 and 50 + 20 doses). Our findings indicate that the mixture of glyphosate and 2,4-D herbicides, mimicking environmentally relevant scenarios, promotes WD-induced changes in the male genital system.

Another Chanarin-Dorfman Syndrome (CDS) in an Egyptian Patient

Abstract Introduction Chanarin–Dorfman syndrome is a very rare autosomal recessive syndrome caused by mutation in ABHD5 a cofactor for adipose triglyceride lipase resulting in intracellular accumulation of triacylglycerol in various body tissues. Several patients were previously reported from Egypt and middle East but still the diagnosis is missed for several years. Here we report another case with this syndrome and describe his journey to diagnosis. Case report A 4.4 year old boy, the fifth in order of birth of first cousin Egyptian parents presented to the Genetics Clinic because of abnormal scaling all over the body since birth. On examination, he had short stature, hepatomegaly, and generalized ichthyosis. His chest, heart and neurological examination showed no abnormalities. Ocular examination revealed refractive errors. Audiometry revealed hearing loss. Peripheral blood smear examination demonstrated cytoplasmic vacuoles within the neutrophils (Jordans bodies ) confirming the diagnosis of CDS. He had family history of sibling death at age of 2 day with extensive scaling overall the body. Conclusion CDS should be suspected in patients with congenital ichthyosis and hepatomegaly. Diagnosis is simple using blood smear to find Jordans bodies.

Administration of probiotics affects Artemia franciscana metanauplii intestinal ultrastructure and offers resistance against a Photobacterium damselae ssp. piscicida induced oxidative stress response

The effects of Photobacterium damselae ssp. piscicida (Phdp) on immune responses and intestinal ultrastructure of Artemia franciscana following infection and their amelioration by the probiotic bacteria Bacillus subtilis, Lactobacillus plantarum and Lactococcus lactis were evaluated. Pathogen growth inhibition in coculture with each probiotic and its virulence against Artemia were confirmed with an LC50 of 105 CFU mL−1. Phdp administration to Artemia at sublethal levels resulted in depletion of superoxide dismutase, glutathione reductase, glutathione transferase and phenoloxidase activities, extensive lipid peroxidation and reduced survival. Following a combined administration of each probiotic and the pathogen, enzyme activities and survival were significantly higher, while lipid peroxidation was reduced, compared to the infected group with no probiotic treatment (P < 0.05). The transmission electron microscopy study revealed that pathogen infection resulted in disarranged and fragmented microvilli, formation of empty or pathogen containing cytoplasmic vacuoles and damaged mitochondria. In the probiotic-treated and Phdp-infected series, intestinal cells showed normal appearance, except for the presence of pathogen-containing vacuoles and highly ordered but laterally stacked microvilli. The results of the present study indicate that Phdp induces cell death through an oxidative stress response and probiotics enhance Artemia immune responses to protect it against the Phdp induced damage.

Discovery of a First-in-Class Degrader for the Lipid Kinase PIKfyve.

The phosphoinositide kinase PIKfyve has emerged as a new potential therapeutic target in various cancers. However, limited clinical progress has been achieved with PIKfyve inhibitors. Here, we report the discovery of a first-in-class PIKfyve degrader 12d (PIK5-12d) by employing the proteolysis-targeting chimera approach. PIK5-12d potently degraded PIKfyve protein with a DC50 value of 1.48 nM and a Dmax value of 97.7% in prostate cancer VCaP cells. Mechanistic studies revealed that it selectively induced PIKfyve degradation in a VHL- and proteasome-dependent manner. PIKfyve degradation by PIK5-12d caused massive cytoplasmic vacuolization and blocked autophagic flux in multiple prostate cancer cell lines. Importantly, PIK5-12d was more effective in suppressing the growth of prostate cancer cells than the parent inhibitor and exerted prolonged inhibition of downstream signaling. Further, intraperitoneal administration of PIK5-12d exhibited potent PIKfyve degradation and suppressed tumor proliferation in vivo. Overall, PIK5-12d is a valuable chemical tool for exploring PIKfyve-based targeted therapy.

Bovine autosomal recessive alpha‐mannosidosis in an Aberdeen Angus herd from Argentina

AbstractAn outbreak of α‐mannosidosis in a bovine herd from Buenos Aires province is reported. Postmortem examination, histology, lectin histochemistry and electron microscopy were performed on five calves born with nervous signs. Hair samples from dams and calves were genetically analysed. The α‐mannosidase gene including exon 7 was amplified and sequenced. At postmortem examination, no gross changes were evident. Histologically, cytoplasmic neuronal vacuolation in the brain and cerebellum of the five calves, staining moderately to intensively, with lectin histochemistry was evident. Ultrastructurally, vacuoles containing finely granulated material in the cytoplasm of neurons, myelin degeneration and axonal swelling were seen in the brain. Genetic analysis revealed a heterozygous carrier state for 961T→C transition in the exon 7 of α‐mannosidosis gene of four cows, and a recessive homozygous state for 961T→C transition in the same genes in the calves analysed. This highlights the importance of identifying and removing animals carrying the α‐mannosidosis mutated gene.

Malignant mesothelioma cells with characteristic intracytoplasmic vacuolization and lipids.

In this brief report, we described some uncommon cytomorphological features of malignant mesothelioma (MM) cells in pleural effusions. The tumor cells exhibited abundant cytoplasmic vacuolization, with presence of single or multiple eccentric nuclei in several cells. In the Giemsa-stained smear, we observed a glossy spherical material in some cells, which tested positive in Sudan III stain. In immunocytochemical analysis, tumor cells were positive for calretinin, podoplanin, epithelial membrane antigen, and methylthioadenosine phosphorylase; tumor cells were negative for BRCA1-associated protein 1, CD68, and desmin. The intracytoplasmic vacuoles were positive for adipophilin expression.

The Effect of Different Concentrations of Bee Venom on Kidney Tissue of Albino Male Rats Rattus

In this study, 60 males of albino rats (Rattus rattus) were tested to study the effect of BV on kidney tissue at two doses of 10 and 40 µg/kg and two times after 10 and 15 days. The sample included 40 rats divided into two groups; 20 rats for each group. These groups were injected intraperitoneally according to the mentioned periods and were considered as a control group. The results exhibited renal tubule dilation and slight atrophy of glomerular tuft in the kidneys of rats treated with 10 µg/kg for 10 days compared to kidneys in the control treatment. Furthermore, 10 µg/kg treatment for 15 days has caused renal tubule dilation and degeneration with cytoplasmic vacuolation. Our findings also showed a slight atrophy of the glomerular tuft as well as a reduction in the number of glomeruli in this subcapsular area. The results also indicate regeneration of kidney tubules in rats given 40 µg/kg for 10 days. Meanwhile, sections of the kidney for rats that were given 40 µg/kg for 15 days exhibited a slight atrophy of glomerular tuft hypertrophied tubular epithelial cells.

Curcumin mediates selective aggregation of mutant p53 in cancer cells: A promising therapeutic strategy

The increased stability of mutant p53 (Mutp53) plays a crucial role in its gain of function, making proteins involved in its stabilization promising targets for drug intervention. Although curcumin is known to exhibit anti-cancer effects, its role as a deubiquitinase (DUB) inhibitor in Mutp53 destabilization remains poorly explored. Our study demonstrates that curcumin treatment induced ubiquitination and destabilization of Mutp53 but not Wild-type p53 (WTp53) in cancer cells. Furthermore, proteasome and lysosome inhibitors failed to reverse the effect of curcumin, indicating Mutp53 destabilization is possibly via an alternate mechanism. Intriguingly, curcumin treatment also resulted in the nuclear aggregation of the Mutp53 protein, which was rescued by combined Dithiothreitol (DTT) treatment. Similar to curcumin, a broad-spectrum deubiquitinase inhibitor induced Mutp53 aggregation implying curcumin possibly acts by inhibiting deubiquitinases. Additionally, curcumin treatment inhibited colony-forming abilities, induced cytoplasmic vacuolation, and cell death selectively in Mutp53-expressing cells. Collectively, our study highlights the potential of curcumin as a promising therapeutic agent for targeting Mutp53-expressing cancer cells.