Inherently Reduced Expression of ASC Restricts Caspase-1 Processing in Hepatocytes and Promotes Plasmodium Infection.

Inflammasome-mediated caspase-1 activation facilitates innate immune control of Plasmodium in the liver, thereby limiting the incidence and severity of clinical malaria. However, caspase-1 processing occurs incompletely in both mouse and human hepatocytes and precludes the generation of mature IL-1ß or IL-18, unlike in other cells. Why this is so or how it impacts Plasmodium control in the liver has remained unknown. We show that an inherently reduced expression of the inflammasome adaptor molecule apoptosis-associated specklike protein containing CARD (ASC) is responsible for the incomplete proteolytic processing of caspase-1 in murine hepatocytes. Transgenically enhancing ASC expression in hepatocytes enabled complete caspase-1 processing, enhanced pyroptotic cell death, maturation of the proinflammatory cytokines IL-1ß and IL-18 that was otherwise absent, and better overall control of Plasmodium infection in the liver of mice. This, however, impeded the protection offered by live attenuated antimalarial vaccination. Tempering ASC expression in mouse macrophages, on the other hand, resulted in incomplete processing of caspase-1. Our work shows how caspase-1 activation and function in host cells are fundamentally defined by ASC expression and offers a potential new pathway to create better disease and vaccination outcomes by modifying the latter.

AIM2 sensors mediate immunity to Plasmodium infection in hepatocytes.

Malaria, caused by Plasmodium parasites is a severe disease affecting millions of people around the world. Plasmodium undergoes obligatory development and replication in the hepatocytes, before initiating the life-threatening blood-stage of malaria. Although the natural immune responses impeding Plasmodium infection and development in the liver are key to controlling clinical malaria and transmission, those remain relatively unknown. Here we demonstrate that the DNA of Plasmodium parasites is sensed by cytosolic AIM2 (absent in melanoma 2) receptors in the infected hepatocytes, resulting in Caspase-1 activation. Remarkably, Caspase-1 was observed to undergo unconventional proteolytic processing in hepatocytes, resulting in the activation of the membrane pore-forming protein, Gasdermin D, but not inflammasome-associated proinflammatory cytokines. Nevertheless, this resulted in the elimination of Plasmodium-infected hepatocytes and the control of malaria infection in the liver. Our study uncovers a pathway of natural immunity critical for the control of malaria in the liver.

The influence of age and sex on the absolute cell numbers of the human brain cerebral cortex.

The human cerebral cortex is one of the most evolved regions of the brain, responsible for most higher-order neural functions. Since nerve cells (together with synapses) are the processing units underlying cortical physiology and morphology, we studied how the human neocortex is composed regarding the number of cells as a function of sex and age. We used the isotropic fractionator for cell quantification of immunocytochemically labeled nuclei from the cerebral cortex donated by 43 cognitively healthy subjects aged 25-87 years old. In addition to previously reported sexual dimorphism in the medial temporal lobe, we found more neurons in the occipital lobe of men, higher neuronal density in women's frontal lobe, but no sex differences in the number and density of cells in the other lobes and the whole neocortex. On average, the neocortex has ~10.2 billion neurons, 34% in the frontal lobe and the remaining 66% uniformly distributed among the other 3 lobes. Along typical aging, there is a loss of non-neuronal cells in the frontal lobe and the preservation of the number of neurons in the cortex. Our study made possible to determine the different degrees of modulation that sex and age evoke on cortical cellularity.

The impact of early sports participation on body fatness in adulthood is not mediated by current physical activity.

OBJECTIVE: The aim was to analyze the relationship between early sports participation (ESP) and body fatness (BF) in adults, as well as to identify whether this possible relationship is directly influenced by the current physical activity (PA) level. METHODS: This cross-sectional study combined baseline data of two cohort. The BF estimated by DXA. The ESP, the subjects reported the engagement in sports during childhood (7-10 years) and adolescence (11-17 years) through two yes/no questions and current PA (described as steps) was device-measured using pedometers. Were identified as potential covariates and therefore adjusted the multivariate models: age, ethnicity, alcohol consumption, smoking, and sleep quality. Statistical analysis consisted of the chi-square test, analysis of variance/covariance, and structural equation modeling (software BioEstat version 5.0; p-value < .05). RESULTS: Adults engaged in ESP had lower BF; among women, the variance in BF explained by ESP was 25.5%; among men, it was 9.2%. Sports participation in early life (r = -.436 [95% CI: -0.527 to -0.346]) and current PA (r = -.431 [95% CI: -0.522 to -0.340]) were inversely related to BF, as well as positively related to each other (r = .328 [95% CI: 0.226 to 0.430]). In the mediation model, current PA partially mediated (18.5%) the impact of ESP on BF, while current PA and ESP remained relevant determinants of BF. CONCLUSION: Early sports participation and current PA have a significant impact on BF in adulthood, which is of similar magnitude and independent of each other.

Assessing pet owners' willingness to embrace insects in pet food.

Alternative diets for pets have gained attention in recent years due to concerns about sustainability and environmental impact. One emerging option is the inclusion of edible insects in pet food. This study aimed to survey dog (DO) and cat (CO) owners regarding their willingness to feed insect-based pet food (IBPF) to their pets and to identify potential motivating factors. A total of 435 valid responses were evaluated, with 66% being DO and 58% CO. Of those, 24% owned both dogs and cats. About half of DO (52%) and 62% of CO were positive about IBPF. CO had a higher acceptance of IBPF than DO (p < 0.05). Moreover, both DO and CO showed a significant increase in acceptance if IBPF has been proven to provide sustainable and nutritional benefits (p < 0.01). CO adhering to a specific diet themselves showed a significantly higher acceptance of IBPF (p < 0.05), however it was independent of the type of diet. Additionally, 32% of DO and 34% of CO reported prior experience with eating edible insects themselves, and a positive entomophagy experience increased DO's (p = 0.108) and CO's (p < 0.01) acceptance of IBPF. Furthermore, the interest in entomophagy resulted in increased acceptance of IBPF by both DO and CO who had not yet tried insects (p < 0.01). In conclusion, pet owners demonstrated a favourable perception of IBPF and showed a willingness to incorporate these products into their pets' diets. CO displayed a more positive perception compared to DO, suggesting a beneficial focus on the feline niche. To enhance overall acceptance, emphasizing sustainability and conducting further research to establish the nutritional benefits of IBPF are crucial. The findings of this study provide valuable insights for pet food manufacturers and marketers to develop strategies and products aligned with the preferences and needs of pet owners.

Full-fat corn germ improves the performance and milk fat yield of Girolando cows fed sugarcane bagasse and cactus cladodes as forage sources.

We investigated the effects of replacing ground corn with full-fat corn germ (FFCG) on milk production, milk composition, and nutrient use in cows fed sugarcane bagasse and cactus cladodes. Ten multiparous Girolando cows (average body weight 500 ± 66 kg, 90 ± 15 days in milk) were distributed in a replicated 5 × 5 Latin Square and assigned to five dietary treatments containing 0%, 25%, 50%, 75%, or 100% of full-fat corn germ in substitution to ground corn. Full-fat corn germ increased fat-corrected milk yield by 2.2 kg/day and the synthesis of fat, lactose, and total solids in milk by 94.4, 60.0, and 201.10 g/day, respectively (p < 0.05). Cows fed corn germ quadratically increased (p < 0.05) dry matter intake by 1.01 kg/day, with the intake of crude protein and total digestible nutrients following the same pattern. Conversely, the substitution of corn for full-fat corn germ linearly reduced (p < 0.05) the total non-fiber carbohydrate intake from 5.79 to 4.40 kg/d. Except for ether extract and non-fiber carbohydrates, full-fat corn germ did not alter (p > 0.05) nutrient digestibility. Cows fed corn germ excreted less (p < 0.05) urea-N in milk and urine N. These results demonstrate that full-fat corn germ can partially replace ground corn to enhance the milk production efficiency of crossbred cows fed cactus cladodes and sugarcane bagasse. Furthermore, including sugarcane bagasse in FFCG-supplemented diets prevents milk fat depression in cows fed cactus cladodes.

Treatment of cutaneous leishmaniasis with a sequential scheme of pentamidine and tamoxifen in an area with a predominance of Leishmania (Viannia) guyanensis: A randomised, non-inferiority clinical trial.

OBJECTIVE: To determine whether a combination of a single intramuscular (IM) dose of pentamidine (7 mg/kg) followed by oral tamoxifen 40 mg/day for 20 days is non-inferior to three IM doses of pentamidine 7 mg/kg in the treatment of cutaneous leishmaniasis with a margin of 15%. METHODS: Phase II, randomised, controlled, open-label, non-inferiority clinical trial. Primary outcome was the complete healing of the lesions 6 months after starting treatment. Secondary outcomes were healing 3 months after starting treatment and determining the presence and severity of adverse effects (AE). RESULTS: The research was concluded with 49 patients; Leishmania (Viannia) guyanensis was the most frequent species isolated. In the primary outcome, 18 (72%) (95% CI: 52.4%-85.7%) of the 25 patients allocated to the intervention group and 24 (100%) (95% CI: 86.2%-100%) of the control group (p = 0.015) met the established criteria of cure. There was no AE with tamoxifen. CONCLUSION: Although a 72% cure rate presented by the combination of tamoxifen and pentamidine was lower than in the control group that achieved a 100% cure, it is still a safe and is a clinically relevant result. It indicates that the therapeutic scheme evaluated may be a promising option for populations in remote areas, however it should be further studied, in order to include a larger number of patients.

Uncovering the Infection Strategy of Phyllachora maydis during Maize Colonization: A Comprehensive Analysis.

Tar spot, a disease caused by the ascomycete fungal pathogen Phyllachora maydis, is considered one of the most significant yield-limiting diseases of maize (Zea mays L.) within the United States. P. maydis may also be found in association with other fungi, forming a disease complex which is thought to result in the characteristic fish eye lesions. Understanding how P. maydis colonizes maize leaf cells is essential for developing effective disease control strategies. Here, we used histological approaches to elucidate how P. maydis infects and multiplies within susceptible maize leaves. We collected tar spot-infected maize leaf samples from four different fields in northern Indiana at three different time points during the growing season. Samples were chemically fixed and paraffin-embedded for high-resolution light and scanning electron microscopy. We observed a consistent pattern of disease progression in independent leaf samples collected across different geographical regions. Each stroma contained a central pycnidium that produced asexual spores. Perithecia with sexual spores developed in the stomatal chambers adjacent to the pycnidium, and a cap of spores formed over the stroma. P. maydis reproductive structures formed around but not within the vasculature. We observed P. maydis associated with two additional fungi, one of which is likely a member of the Paraphaeosphaeria genus; the other is an unknown fungi. Our data provide fundamental insights into how this pathogen colonizes and spreads within maize leaves. This knowledge can inform new approaches to managing tar spot, which could help mitigate the significant economic losses caused by this disease.

Full-fat corn germ in diets for dairy cows as an alternative to ground corn.

The experiments reported in this research paper address the effects of replacing ground corn (GC) with full-fat corn germ (FFCG) on nutrient intake and digestibility, nitrogen utilization efficiency, performance, and predicted methane production in dairy cows fed cactus cladodes and sugarcane. We hypothesized that the inclusion of FFCG in the diet would not alter the performance of lactating cows but would reduce the predicted methane production in vivo. Ten multiparous Holstein cows at 90 ± 10 d of lactation and yielding 24.2 ± 3.5 kg milk/d were assigned to dietary treatments consisting of different levels of replacement of GC by FFCG (0; 25; 50; 75 and 100% of diet dry matter) in a replicated 5 × 5 Latin square design with 21-d periods. Methane production was predicted using an automated gas in vitro production system. Except for ether extract intake, which increased, the intake of all nutrients decreased linearly with the replacement of GC by FFCG. The digestibility of dry matter, organic matter and neutral detergent fiber reduced, whereas the digestibility of ether extract increased linearly with FFCG. There were no changes in the digestibility of crude protein. The nitrogen intake and daily excretion in urine and feces decreased, while nitrogen use efficiency increased linearly. There was no significant effect of diets on nitrogen balance or microbial protein synthesis and efficiency. The yield of protein, lactose and total solids in milk showed a quadratic behavior. On the other hand, milk fat yield and energy-corrected milk yield decreased linearly with the replacement of GC by FFCG. No effect on pH or ammonia nitrogen was observed. The production of methane (CH4, g/kg DM) and total CH4 (g/d), and CH4 intensity decreased linearly with the replacement of GC by FFCG. In conclusion, FFCG has been shown to be an effective source of fat to reduce methane production in dairy cows, partially supporting our initial hypothesis. However, as it decreases milk fat production, it is not recommended to replace more than 50% of GC by FFCG for lactating cows fed cactus cladodes and sugarcane.

COPD Patients Exhibit Distinct Gene Expression, Accelerated Cellular Aging, and Bias to M2 Macrophages.

COPD, one of world's leading contributors to morbidity and mortality, is characterized by airflow limitation and heterogeneous clinical features. Three main phenotypes are proposed: overlapping asthma/COPD (ACO), exacerbator, and emphysema. Disease severity can be classified as mild, moderate, severe, and very severe. The molecular basis of inflammatory amplification, cellular aging, and immune response are critical to COPD pathogenesis. Our aim was to investigate EP300 (histone acetylase, HAT), HDAC 2 (histone deacetylase), HDAC3, and HDAC4 gene expression, telomere length, and differentiation ability to M1/M2 macrophages. For this investigation, 105 COPD patients, 42 smokers, and 73 non-smoker controls were evaluated. We identified a reduced HDAC2 expression in patients with mild, moderate, and severe severity; a reduced HDAC3 expression in patients with moderate and severe severity; an increased HDAC4 expression in patients with mild severity; and a reduced EP300 expression in patients with severe severity. Additionally, HDAC2 expression was reduced in patients with emphysema and exacerbator, along with a reduced HDAC3 expression in patients with emphysema. Surprisingly, smokers and all COPD patients showed telomere shortening. COPD patients showed a higher tendency toward M2 markers. Our data implicate genetic changes in COPD phenotypes and severity, in addition to M2 prevalence, that might influence future treatments and personalized therapies.

Optimization Studies and Compositional Oil Analysis of Pequi (Caryocar brasiliense Cambess) Almonds by Supercritical CO2 Extraction.

Caryocar brasiliense Cambess (pequi) is the fruit of the pequizeiro tree found in the Brazilian Cerrado (savanna). Supercritical fluids have been used to effectively extract bioactive chemicals. In light of the paucity of research on the supercritical extraction of pequi, in this study, experimental tests were conducted on the extraction of pequi almond oil using supercritical CO2; the optimal extraction conditions were determined, and the fatty acids and active compounds in the oil were characterized. The experiments were conducted using the Box-Behnken experimental design of a three-variable system: pressure (15, 20, and 25 MPa), temperature (303.15, 318.15, and 333.15 K), and flow rate (2, 3, and 5 g.min-1). The optimal extraction conditions were 318.15 K, 25 MPa, and 5.0 g.min-1, which yielded 27.6 wt% of oil. The experimental kinetic curves were described using a second-order quadratic model (based on the Sovová model), which demonstrated a satisfactory correspondence with the kinetic curves. Significant amounts of squalene, stigmasterol, oleic fatty acids, and palmitic fatty acids were detected in pequi almond oil.

Parental beliefs in and attitudes toward teething signs and symptoms: A systematic review.

BACKGROUND: Parents believe that teething is associated with signs and symptoms, which may induce them to give medications that could harm their children. Some children may require alleviation of symptoms and overall attention. AIM: To assess parents' beliefs in and attitudes toward teething. DESIGN: Through electronic databases and gray literature, this systematic review identified cross-sectional studies reporting parents' beliefs in, knowledge about, and attitudes toward the signs and symptoms of primary tooth eruption in children aged between 0 and 36 months. Three reviewers independently selected the studies, collected the information, assessed methodological quality, and checked for accuracy with disagreements solved by a fourth reviewer. The Agency of Research and Quality in Health questionnaire for cross-sectional studies was used for quality assessment. Descriptive analysis with median and interquartile ranges was adopted. RESULTS: Twenty-nine studies comprising 10 524 participants from all geographic regions were included. The methodological quality of the studies was moderate. Most parents have beliefs in signs and symptoms during dentition, the most reported symptom being the desire to bite. Oral rehydration was the most exposed attitude in the studies included. Only a small proportion of parents reported no attitude. CONCLUSIONS: The majority of parents believed in at least one sign or symptom associated with teething, and only few of them would do nothing or just wait for the signs or symptoms to pass, with no difference among countries (Protocol doi: 10.17605/OSF.IO/S2KZ3).

Can phytogenic additives replace monensin sodium in beef cattle feeding?

Monensin sodium is one of the most common ionophores used in livestock feeding; however, ionophores are condemned by organized consumer groups. Bioactive compounds from plants found in the seasonally dry tropical forest have similar mechanisms of action as ionophores. The aim was to investigate the effects of replacing monensin sodium with phytogenic additives on the nutritional efficiency of beef cattle. Five 14-month-old Nellore bulls (average body weight 452.68 ± 42.60 kg) were used in the study. The experiment was designed as a 5 × 5 Latin Square (five treatments and five 22-day experimental periods). Within each period, 15 days were used for adaptation of animals to experimental conditions and 7 days for data collection. Bulls were fed a control diet (without additives), monensin (a diet containing monensin sodium 40% as a synthetic additive), and three diets containing phytogenic additives prepared from Anadenanthera macrocarpa (Benth) Brenan, Mimosa tenuiflora (Willd) Poiret, or Prosopis juliflora (Sw.) DC. Nutritional efficiency was assessed through feed intake, nutrient digestibility, feeding behavior, and hematological parameters. Monensin and phytogenic additives did not influence (P > 0.05) feeding behavior or hematological parameters, but the nutrient intake was highest for bulls supplemented phytogenic additives (P < 0.05). Monensin supplementation did not influence (P > 0.05) feed intake. The phytogenic additives and monensin sodium increased (P < 0.05) the nutrient digestibility. Therefore, the phytogenic additives from P. juliflora, A. macrocarpa, and M. tenuiflora can be recommended to enhance the nutritional efficiency of confined Nellore cattle.

Relocating to a new facility: The challenge of a cord blood banking transferral in Brazil.

BACKGROUND: Our cord blood banking facility planned and executed a transferral of its entire operation to a new site in the South of Brazil. Transporting LN2 freezers is a complex process in which extensive planning is essential to minimize the risks of damaging products or storage units. METHODS: To fulfill this objective, we constructed a detailed relocation plan consisting of four phases and risk mitigation measures, collaborated with the representatives of all departments, regulating agencies, and professionals from the transport company, and assembled a validation plan for cryogenic freezers and the viability of cord blood units. RESULTS: The new facility was prepared in accordance with the project plan, local legislation, quality system program requirements, and accreditation agency guidance. A 12-h operation of moving the cryogenic freezers was conducted successfully, with no loss or damage of client samples or equipment. CONCLUSION: Through the development and execution of a transferral plan, the engagement of appropriate partners, and compliance with security measures from health and government agencies, a successful transferral of a cord blood banking facility operation in its entirety can be successfully accomplished.

Phytogenic additive from Prosopis juliflora on populations of rumen ciliate protozoa and its correlation with nutrition of sheep.

We evaluated the effect of phytogenic additive Prosopis juliflora on populations of ruminal ciliated protozoa and its correlation with variables related to sheep nutrition. In this experiment, five cannulated adult Santa Ines ewes were submitted to the additive intake. Each animal received 6 mL of the extract daily. The experimental design adopted was the Latin square 5 × 5 (five concentrations of additive 0, 200, 400, 600 and 800 mg/mL of water and five periods of 18 days). The additive quadratically reduces the number of large, small and total ciliating protozoa, without promoting changes in the number of medium protozoa in the rumen. Among the genera investigated, Isotricha and Dasytricha were the only ones affected by supplementation. Additionally, the concentration of ruminal protozoa correlates significantly with the variables related to the nutrition of the animal. The effects of the additive on these variables were particularly pronounced at concentrations around 600 mg/mL.

Identification of Leishmania species by next generation sequencing of hsp70 gene.

Leishmaniasis is a major public health problem worldwide. Although next generation sequencing technology has been widely used in the diagnosis of infectious diseases, it has been scarcely applied in identification of Leishmania species. The aim of this study was to compare the efficiency of MinION™ nanopore sequencing and polymerase chain reaction restriction fragment length polymorphism in identifying Leishmania species. Our results showed that the MinION™ sequencer was able to discriminate reference strains and clinical samples with high sensitivity in a cost and time effective manner without the prior need for culture.

Human adipose-derived mesenchymal stromal cells from face and abdomen undergo replicative senescence and loss of genetic integrity after long-term culture.

Body fat depots are heterogeneous concerning their embryonic origin, structure, exposure to environmental stressors, and availability. Thus, investigating adipose-derived mesenchymal stromal cells (ASCs) from different sources is essential to standardization for future therapies. In vitro amplification is also critical because it may predispose cell senescence and mutations, reducing regenerative properties and safety. Here, we evaluated long-term culture of human facial ASCs (fASCs) and abdominal ASCs (aASCs) and showed that both met the criteria for MSCs characterization but presented differences in their immunophenotypic profile, and differentiation and clonogenic potentials. The abdominal tissue yielded more ASCs, and these had higher proliferative potential, but facial cells displayed fewer mitotic errors at higher passages. However, both cell types reduced clonal efficiency over time and entered replicative senescence around P12, as evaluated by progressive morphological alterations, reduced proliferative capacity, and SA-ß-galactosidase expression. Loss of genetic integrity was detected by a higher proportion of cells showing nuclear alterations and γ-H2AX expression. Our findings indicate that the source of ASCs can substantially influence their phenotype and therefore should be carefully considered in future cell therapies, avoiding, however, long-term culture to ensure genetic stability.

Impact of JAK/STAT inhibitors on human monocyte-derived-macrophages stimulated by cigarette smoke extract and lipopolysaccharide.

The main risk factor for chronic obstructive pulmonary disease (COPD) is cigarette smoke (CS). It can alter many immune cells functions such as phagocytosis, efferocytosis and cytokine production. Cytokines play a role in the orchestration of inflammation in COPD. The JAK/STAT pathways are among the most important signalling components of cytokines. The objective of this work was to investigate the role of the JAK/STAT pathway with regard to cytokine release and microsphere uptake capacity (to minimize the non-specific scavenging) in human monocyte-derived-macrophages (MDMs). The MDMs were stimulated by cigarette smoke extract (CSE) alone or in combination with lipopolysaccharide (LPS). CSE alone was not associated with significant changes in the cytokine, with the exception of IL-8/CXCL8 production. However, CSE disturbed cytokine production in LPS-stimulated MDMs. CSE increase CXCL-8 and CCL2 release in LPS-stimulated monocyte-derived macrophages and suppressed the production of IL-6 and CXCL1 in these cells. CSE also decreased microsphere uptake capacity by MDMs. Then, CSE + LPS-stimulated MDMs were treated with two different JAK inhibitors. AG490 (specific inhibitor of JAK2) and ruxolitinib (inhibitor of JAK1 and JAK2). JAK/STAT inhibitors, particularly ruxolitinib, attenuated in cytokine production without completely inhibiting when compared with dexamethasone. On the other hand, the cells exposed to dexamethasone are nearly unable to capture the microspheres, while both JAK inhibitors do not affect the uptake capacity. In summary, our results showed the versatility of ruxolitinib which might bring a better balance disturbance of cytokine release and uptake capacity. The information regarding the distinctive effect of JAK/STAT inhibitors may be useful in the development of novel treatments for COPD.

"The COVID 19 pandemic worsened my living and working conditions". A qualitative research study of female Brazilian immigrants in Oporto.

Immigrant women face a double vulnerability, being a woman and an immigrant, a situation that hinders their entry into the labor market and worsens their working conditions and occupational health with respect to the native population. The objective of these women is to seek employment or educational opportunities and improving their own lives and those of their families. All this is affected by the axes of gender, ethnicity, migration and socioeconomic level, as well as being confronted with the patriarchal structure of exercise of power and domination. These situations of inequality have aggravated due to the COVID-19 pandemic, worsening working conditions and their occupational health. This study aims to explore the working conditions of the female Brazilian immigrant population living in Porto (Portugal) and how these conditions may affect their health. Qualitative research through semi-structured interviews conducted during the second wave of the COVID-19 pandemic was used. The results show of them had work overload, manifesting anxiety and stress. Physical affectations related to poor work ergonomics and the lack of occupational health examinations in working immigrant women is highlighted. The importance of strengthening migration policies related to occupational health is highlighted. In pandemic situations, the vulnerability of these women increases, worsening their overall health.

Cellular Model of Malignant Transformation of Primary Human Astrocytes Induced by Deadhesion/Readhesion Cycles.

Astrocytoma is the most common and aggressive tumor of the central nervous system. Genetic and environmental factors, bacterial infection, and several other factors are known to be involved in gliomagenesis, although the complete underlying molecular mechanism is not fully understood. Tumorigenesis is a multistep process involving initiation, promotion, and progression. We present a human model of malignant astrocyte transformation established by subjecting primary astrocytes from healthy adults to four sequential cycles of forced anchorage impediment (deadhesion). After limiting dilution of the surviving cells obtained after the fourth deadhesion/readhesion cycle, three clones were randomly selected, and exhibited malignant characteristics, including increased proliferation rate and capacity for colony formation, migration, and anchorage-independent growth in soft agar. Functional assay results for these clonal cells, including response to temozolomide, were comparable to U87MG-a human glioblastoma-derived cell lineage-reinforcing malignant cell transformation. RNA-Seq analysis by next-generation sequencing of the transformed clones relative to the primary astrocytes revealed upregulation of genes involved in the PI3K/AKT and Wnt/ß-catenin signaling pathways, in addition to upregulation of genes related to epithelial-mesenchymal transition, and downregulation of genes related to aerobic respiration. These findings, at a molecular level, corroborate the change in cell behavior towards mesenchymal-like cell dedifferentiation. This linear progressive model of malignant human astrocyte transformation is unique in that neither genetic manipulation nor treatment with carcinogens are used, representing a promising tool for testing combined therapeutic strategies for glioblastoma patients, and furthering knowledge of astrocytoma transformation and progression.