Evaluating responses by ChatGPT to farmers' questions on irrigated lowland rice cultivation in Nigeria.

The limited number of agricultural extension agents (EAs) in sub-Saharan Africa limits farmers' access to extension services. Artificial intelligence (AI) assistants could potentially aid in providing answers to farmers' questions. The objective of this study was to evaluate the ability of an AI chatbot assistant (ChatGPT) to provide quality responses to farmers' questions. We compiled a list of 32 questions related to irrigated rice cultivation from farmers in Kano State, Nigeria. Six EAs from the state were randomly selected to answer these questions. Their answers, along with those of ChatGPT, were assessed by four evaluators in terms of quality and local relevancy. Overall, chatbot responses were rated significantly higher quality than EAs' responses. Chatbot responses received the best score nearly six times as often as the EAs' (40% vs. 7%). The evaluators preferred chatbot responses to EAs in 78% of cases. The topics for which the chatbot responses received poorer scores than those by EAs included planting time, seed rate, and fertilizer application rate and timing. In conclusion, while the chatbot could offer an alternative source for providing agricultural advisory services to farmers, incorporating site-specific input rate-and-timing agronomic practices into AI assistants is critical for their direct use by farmers.

Inorganic fertilizer use and its association with rice yield gaps in sub-Saharan Africa.

Where and which countries should receive higher priority for improving inorganic fertilizer use in rice fields in sub-Saharan Africa (SSA)? This study addressed this question by assessing the spatial variation in fertilizer use and its association with rice yield and yield gap in 24 SSA countries through a systematic literature review of peer-reviewed papers, theses, and grey literature published between 1995 and 2021. The results showed a large variation in N, P, and K fertilizer application rates and rice yield and an opportunity for narrowing the yield gap by increasing N and P rates, especially in irrigated rice systems. We identified clusters of sites/countries based on nutrient input and yield and suggested research and development strategies for improving yields and optimizing nutrient use efficiencies. Further research is essential to identify the factors causing low fertilizer use and the poor association between its use and yield in rainfed systems.

Estimating nutrient concentrations and uptake in rice grain in sub-Saharan Africa using linear mixed-effects regression.

Context or problem: Quantification of nutrient concentrations in rice grain is essential for evaluating nutrient uptake, use efficiency, and balance to develop fertilizer recommendation guidelines. Accurate estimation of nutrient concentrations without relying on plant laboratory analysis is needed in sub-Saharan Africa (SSA), where farmers do not generally have access to laboratories. Objective or research question: The objectives are to 1) examine if the concentrations of macro- (N, P, K, Ca, Mg, S) and micronutrients (Fe, Mn, B, Cu) in rice grain can be estimated using agro-ecological zones (AEZ), production systems, soil properties, and mineral fertilizer application (N, P, and K) rates as predictor variables, and 2) to identify if nutrient uptakes estimated by best-fitted models with above variables provide improved prediction of actual nutrient uptakes (predicted nutrient concentrations x grain yield) compared to average-based uptakes (average nutrient concentrations in SSA x grain yield). Methods: Cross-sectional data from 998 farmers' fields across 20 countries across 4 AEZs (arid/semi-arid, humid, sub-humid, and highlands) in SSA and 3 different production systems: irrigated lowland, rainfed lowland, and rainfed upland were used to test hypotheses of nutrient concentration being estimable with a set of predictor variables among above-cited factors using linear mixed-effects regression models. Results: All 10 nutrients were reasonably predicted [Nakagawa's R2 ranging from 0.27 (Ca) to 0.79 (B), and modeling efficiency ranging from 0.178 (Ca) to 0.584 (B)]. However, only the estimation of K and B concentrations was satisfactory with a modeling efficiency superior to 0.5. The country variable contributed more to the variation of concentrations of these nutrients than AEZ and production systems in our best predictive models. There were greater positive relationships (up to 0.18 of difference in correlation coefficient R) between actual nutrient uptakes and model estimation-based uptakes than those between actual nutrient uptakes and average-based uptakes. Nevertheless, only the estimation of B uptake had significant improvement among all nutrients investigated. Conclusions: Our findings suggest that with the exception of B associated with high model EF and an improved uptake over the average-based uptake, estimates of the macronutrient and micronutrient uptakes in rice grain can be obtained simply by using average concentrations of each nutrient at the regional scale for SSA. Implications: Further investigation of other factors such as the timing of fertilizer applications, rice variety, occurrence of drought periods, and atmospheric CO2 concentration is warranted for improved prediction accuracy of nutrient concentrations.

Deep Learning Enables Instant and Versatile Estimation of Rice Yield Using Ground-Based RGB Images.

Rice (Oryza sativa L.) is one of the most important cereals, which provides 20% of the world's food energy. However, its productivity is poorly assessed especially in the global South. Here, we provide a first study to perform a deep-learning-based approach for instantaneously estimating rice yield using red-green-blue images. During ripening stage and at harvest, over 22,000 digital images were captured vertically downward over the rice canopy from a distance of 0.8 to 0.9 m at 4,820 harvesting plots having the yield of 0.1 to 16.1 t·ha-1 across 6 countries in Africa and Japan. A convolutional neural network applied to these data at harvest predicted 68% variation in yield with a relative root mean square error of 0.22. The developed model successfully detected genotypic difference and impact of agronomic interventions on yield in the independent dataset. The model also demonstrated robustness against the images acquired at different shooting angles up to 30° from right angle, diverse light environments, and shooting date during late ripening stage. Even when the resolution of images was reduced (from 0.2 to 3.2 cm·pixel-1 of ground sampling distance), the model could predict 57% variation in yield, implying that this approach can be scaled by the use of unmanned aerial vehicles. Our work offers low-cost, hands-on, and rapid approach for high-throughput phenotyping and can lead to impact assessment of productivity-enhancing interventions, detection of fields where these are needed to sustainably increase crop production, and yield forecast at several weeks before harvesting.

Remineralizing Potential of Natural Nano-Hydroxyapatite Obtained from Epinephelus chlorostigma in Artificially Induced Early Enamel Lesion: An In Vitro Study.

Dental caries is a common problem in adolescents, leading to permanent loss of teeth or cavitation. Caries is a continuous process wherein demineralization and remineralization occur regularly. Hydroxyapatite (HA) is one of the most biocompatible and bioactive materials, as it closely resembles the mineral composition of teeth. The present study deals with isolating hydroxyapatite from fish bone (Epinephelus chlorostigma) by alkaline hydrolysis and thermal calcination. The isolated nano HA was characterized using FT-IR, XRD, TGA, FE-SEM-EDX, and HR-TEM analysis. The nano HA isolated by alkaline hydrolysis is nontoxic, and the cells are viable. The isolated HA enhances the proliferation of L929 cells. The remineralization potential of the extracted nano HA was evaluated in healthy premolars by DIAGNOdent/laser fluorescence quantification, surface microhardness test, and SEM-EDX analysis. Surface morphological observations in SEM and EDX analyses show that thermally calcined HA and alkali-treated HA can induce mineralization and deposit minerals. Therefore, HA obtained from Epinephelus chlorostigma could be a potential biomaterial for treating early caries.

Anatomical root responses of rice to combined phosphorus and water stress - relations to tolerance and breeding opportunities.

Drought and low P availability are major limitations for rainfed rice (Oryza spp.) production. Root anatomy plays a key role in resource acquisition and tolerance to P and water limitations. Root anatomical responses of three contrasting rice varieties to combinations of different levels of P (deficient to non-limiting) and water availability (water stress to submergence) were evaluated in two pot trials. P availability was the dominant growth-limiting factor, but anatomical root responses to water availability were more prominent than responses to P availability. Cortical cell file number and number of xylem vessels decreased as a response to water stress, but stele and xylem diameter increased. Low P availability induced thinner xylem vessels and a thinner stele. Drought tolerance related to an overall thicker root stele, thicker xylem vessels and a larger water conductance. Some root traits were observed to be more responsive to water and P availability, whereas other traits were more robust to these environmental factors but highly determined by variety. The observed genotypic variation in root anatomy provides opportunities for trait-based breeding. The plasticity of several traits to multiple environmental factors highlights the need for strategic trait selection or breeding adapted to specific target environments.

Genotypic Variation in Grain P Loading across Diverse Rice Growing Environments and Implications for Field P Balances.

More than 60% of phosphorus (P) taken up by rice (Oryza spp.) is accumulated in the grains at harvest and hence exported from fields, leading to a continuous removal of P. If P removed from fields is not replaced by P inputs then soil P stocks decline, with consequences for subsequent crops. Breeding rice genotypes with a low concentration of P in the grains could be a strategy to reduce maintenance fertilizer needs and slow soil P depletion in low input systems. This study aimed to assess variation in grain P concentrations among rice genotypes across diverse environments and evaluate the implications for field P balances at various grain yield levels. Multi-location screening experiments were conducted at different sites across Africa and Asia and yield components and grain P concentrations were determined at harvest. Genotypic variation in grain P concentration was evaluated while considering differences in P supply and grain yield using cluster analysis to group environments and boundary line analysis to determine minimum grain P concentrations at various yield levels. Average grain P concentrations across genotypes varied almost 3-fold among environments, from 1.4 to 3.9 mg g-1. Minimum grain P concentrations associated with grain yields of 150, 300, and 500 g m-2 varied between 1.2 and 1.7, 1.3 and 1.8, and 1.7 and 2.2 mg g-1 among genotypes respectively. Two genotypes, Santhi Sufaid and DJ123, were identified as potential donors for breeding for low grain P concentration. Improvements in P balances that could be achieved by exploiting this genotypic variation are in the range of less than 0.10 g P m-2 (1 kg P ha-1) in low yielding systems, and 0.15-0.50 g P m-2 (1.5-5.0 kg P ha-1) in higher yielding systems. Improved crop management and alternative breeding approaches may be required to achieve larger reductions in grain P concentrations in rice.

Enhancement of human sodium iodide symporter gene therapy for breast cancer by HDAC inhibitor mediated transcriptional modulation.

The aberrant expression of human sodium iodide symporter (NIS) in breast cancer (BC) has raised the possibility of using targeted radioiodide therapy. Here we investigate modulation of endogenous, functional NIS expression by histone deacetylase inhibitors (HDACi) in vitro and in vivo. Luciferase reporter based initial screening of six different HDACi shows 2-10 fold enhancement of NIS promoter activity in majority of the cell types tested. As a result of drug treatment, endogenous NIS transcript and protein shows profound induction in BC cells. To get an insight on the mechanism of such transcriptional activation, role of Stat4, CREB and other transcription factors are revealed by transcription factor profiling array. Further, NIS-mediated intracellular iodide uptake also enhances substantially (p < 0.05) signifying functional relevance of the transcriptional modulation strategy. Gamma camera imaging confirms 30% higher uptake in VPA or NaB treated BC tumor xenograft. Corroborating with such functional impact of NIS, significant reduction in cell survival (p < 0.005) is observed in VPA, NaB or CI994 drug and (131)I combination treatment in vivo indicating effective radioablation. Thus, for the first time this study reveals the mechanistic basis and demonstrates functional relevance of HDACi pre-treatment strategy in elevating NIS gene therapy approach for BC management in clinic.

Anticancer effects of fucoidan.

Recently, there has been an increased interest in the pharmacologically active natural compounds isolated and used for remedies of various kinds of diseases, including cancer. The great deal of interest has been developed to isolate bioactive compounds from marine resources because of their numerous health beneficial effects. Among marine resources, marine algae are valuable sources of structurally diverse bioactive compounds. Fucoidan is a sulfated polysaccharide derived from brown seaweeds and has been used as an ingredient in some dietary supplement products. Fucoidan has various biological activities including antibacterial, antioxidant, anti-inflammatory, anticoagulant, and antitumor activities. So this chapter deals with anticancer effects of fucoidan.

Anti-proliferative and apoptosis inducing effect of nimbolide by altering molecules involved in apoptosis and IGF signalling via PI3K/Akt in prostate cancer (PC-3) cell line.

Prostate cancer is responsible for major deaths globally after lung cancer. Nimbolide is an important constituent of neem, and it acts as a potent inhibitor for many cancer cells. The present study was designed to evaluate the effects of nimbolide on apoptosis and insulin-like growth factor (IGF) signalling molecules in androgen-independent prostate cancer (PC-3) cells line. Nimbolide (0.5-2 µM) treatment resulted in 50% inhibition at a dose of 2 µM in the PC-3 cell line. The mRNA expression of Fas ligand, Fas-associated death domain receptor (FADDR), Bcl-2-associated X protein (Bax), Bcl-2-associated death promoter (Bad), phosphatidylinositide 3-kinases (PI3K), Akt, IGF1, IGF1 receptor (IGF1R) and IGF binding protein 3 were quantified by reverse transcription polymerase chain reaction and protein expression of Bax, cytochrome c, X-linked inhibitor of apoptosis protein (XIAP), B-Cell Lymphoma 2 (Bcl-2), caspases -8, -9, -10 and -3, poly(ADP-ribose) polymerase (PARP), cleaved PARP, IGF1R, PI3K, Akt, p-Akt was determined by western blot analysis, in nimbolide-treated PC-3 cell line. Nimbolide-induced apoptosis by activating DNA fragmentation in PC-3 cells. Nimbolide treatment increased the mRNA of Fas ligand, FADDR, Bax, Bad and IGF binding protein 3, decreased PI3K, Akt, IGF1 and IGF1R, increased protein expression of caspases 8, 3, 10, 9, Bax and cytochrome c and decreased the expression of XIAP, Bcl2, cleaved PARP, p-Akt and IGF1R. The results suggest that nimbolide acts as a potent anti-cancer agent by inducing apoptosis and inhibiting cell proliferation via PI3K/Akt pathway in PC-3 cells.

Chemopreventive effect of quercetin in MNU and testosterone induced prostate cancer of Sprague-Dawley rats.

Prostate cancer becomes an ideal target for chemoprevention because of its high incidence and extended natural history. The consumption of quercetin (plant flavonoid) in diet is associated with decreased risk of disease and many cancers but then this was not elucidated in prostate malignancy. Hence, a study in which the male Sprague-Dawley rats were induced prostate cancer by hormone (testosterone) and carcinogen (MNU) and simultaneously supplemented with quercetin (200 mg/Kg body weight) thrice a week, was conducted. After the treatment period, rats were killed; ventral and dorsolateral lobes of the prostate were dissected. Histology and oxidative stress markers LPO, H2O2, and antioxidant GSH level were measured in both lobes. The lipid peroxidation, H2O2, in (MNU+T) treated rats were increased and GSH level was decreased, whereas simultaneous quercetin-treated rats reverted back to normal level in both ventral and dorsolateral regions. The different patterns of PIN were observed with associated hyperplasia and dysplasia; changes in these regions and the occurrence of this lesion were reduced in simultaneous quercetin-treated rats. The study concluded that dietary quercetin prevented MNU + T-induced prostate carcinogenesis on both ventral and dorsolateral lobes of Sprague-Dawley rats.

Antiangiogenic effects of marine sponge derived compounds on cancer.

The term "angiogenic switch" refers to a time-restricted event during tumor progression where the balance between pro- and anti-angiogenic factors, resulting in the transition from dormant avascularized hyperplasia to outgrowing vascularized tumor and eventually to malignant tumor progression. Targeting angiogenesis and its mechanistic pathways are critical target for cancer therapy. Recently, marine derived compounds, plays major role in cancer research. Several sponge derived compounds such as alkaloids, terpenes, macrocylic lactone and polyketide are leading drugs in the treatment of different types of diseases including cancer. Those marine sponge compounds inhibit cancer cell proliferation and tumor angiogenesis. Hence, this review sheds light on angiogenic regulators and marine sponge derived antiangiogenic compounds for cancer.

Biosynthesis, antimicrobial and cytotoxic effect of silver nanoparticles using a novel Nocardiopsis sp. MBRC-1.

The biosynthesis of nanoparticles has been proposed as a cost effective environmental friendly alternative to chemical and physical methods. Microbial synthesis of nanoparticles is under exploration due to wide biomedical applications, research interest in nanotechnology and microbial biotechnology. In the present study, an ecofriendly process for the synthesis of nanoparticles using a novel Nocardiopsis sp. MBRC-1 has been attempted. We used culture supernatant of Nocardiopsis sp. MBRC-1 for the simple and cost effective green synthesis of silver nanoparticles. The reduction of silver ions occurred when silver nitrate solution was treated with the Nocardiopsis sp. MBRC-1 culture supernatant at room temperature. The nanoparticles were characterized by UV-visible, TEM, FE-SEM, EDX, FTIR, and XRD spectroscopy. The nanoparticles exhibited an absorption peak around 420 nm, a characteristic surface plasmon resonance band of silver nanoparticles. They were spherical in shape with an average particle size of 45 ± 0.15 nm. The EDX analysis showed the presence of elemental silver signal in the synthesized nanoparticles. The FTIR analysis revealed that the protein component in the form of enzyme nitrate reductase produced by the isolate in the culture supernatant may be responsible for reduction and as capping agents. The XRD spectrum showed the characteristic Bragg peaks of 1 2 3, 2 0 4, 0 4 3, 1 4 4, and 3 1 1 facets of the face centered cubic silver nanoparticles and confirms that these nanoparticles are crystalline in nature. The prepared silver nanoparticles exhibited strong antimicrobial activity against bacteria and fungi. Cytotoxicity of biosynthesized AgNPs against in vitro human cervical cancer cell line (HeLa) showed a dose-response activity. IC50 value was found to be 200 µg/mL of AgNPs against HeLa cancer cells. Further studies are needed to elucidate the toxicity and the mechanism involved with antimicrobial and anticancer activity of the synthesized AgNPs as nanomedicine.

Brown seaweed fucoidan: biological activity and apoptosis, growth signaling mechanism in cancer.

Seaweeds, being abundant sources of bioactive components have much interest in recent times. The complex polysaccharides from the brown, red and green seaweeds possess broad spectrum therapeutic properties. The sulfated polysaccharides are routinely used in biomedical research and have known biological activities. Fucoidan, a fucose-rich polysaccharide extracted from brown seaweed has various biological functions including anticancer effects. Cellular damage induces growth arrest and tumor suppression by inducing apoptosis, the mechanism of cell death depends on the magnitude of DNA damage following exposure to anticancer agents. Apoptosis is mainly regulated by cell growth signaling molecules. Number of research studies evidenced that fucoidan shown to induce cytotoxicity of various cancer cells, induces apoptosis, and inhibits invasion, metastasis and angiogenesis of cancer cells. There are few articles discussing on fucoidan biological activity but no specific review on cancer and its signaling mechanism. Hence, this review discusses the brown seaweed fucoidan structure and some biological function and role in apoptosis, invasion, metastasis, angiogenesis and growth signal mechanism on cancer.

Isolation and characterization of biologically active melanin from Actinoalloteichus sp. MA-32.

Melanins are enigmatic pigments and biological macromolecules that are produced by a wide variety of microorganisms including several species of bacteria and fungi. The present study was carried out on isolation and characterization of melanin from marine actinobacteria, Actinoalloteichus sp. MA-32. Medium composition and culture conditions for the melanin production by Actinoalloteichus sp. MA-32 were optimized using two statistical methods: Plackett-Burman design applied to find the key ingredients and conditions for the best yield of melanin production and central composite design used to optimize the concentration of the four significant variables: glycerol, L-tyrosine, NaCl and trace salt solution. The melanin was optimally active at pH 7-9 and temperature 45-60°C and it was most stable up to pH 11 and 4% of NaCl concentration. Melanin was examined by UV-vis absorption spectroscopy and infrared spectrometry. Melanin has potential antibacterial activity as it showed greater antagonistic and it has a strong antioxidant potential observed in the in vitro evaluation of its DPPH radical-scavenging activity, superoxide radical-scavenging activity, nitric oxide-scavenging activity, reducing power and metal chelating activity. The observed activities indicate that melanin might be a novel potential antioxidant. This study suggested that the melanin could potentially be used as a natural antioxidant in the food, cosmetic and pharmaceutical industries.

Production and characterization of an extracellular polysaccharide from Streptomyces violaceus MM72.

The isolation, optimization, purification and characterization of an extracellular polysaccharide (EPS) from a marine actinobacterium, Streptomyces violaceus MM72 were investigated. Medium composition and culture conditions for the EPS production by S. violaceus MM72 were optimized using two statistical methods: Plackett-Burman design applied to find the key ingredients and conditions for the best yield of EPS production and central composite design used to optimize the concentration of the three significant variables: glucose, tryptone and NaCl. The preferable culture conditions for EPS production were pH 7.0, temperature 35°C and NaCl concentration 2.0% for 120h with fructose and yeast extract as best carbon and nitrogen sources, respectively. The results showed that S. violaceus MM72 produced a kind of EPS having molecular weight of 8.96×10(5)Da. In addition, the EPS showed strong DPPH radical-scavenging activity, superoxide scavenging and metal chelating activities while moderate inhibition of lipid peroxidation and reducing activities determined in this study. These results showed the great potential of EPS produced by S. violaceus MM72 could be used in industry in place of synthetic compounds. The EPS from S. violaceus MM72 may be a new source of natural antioxidants with potential value for health, food and therapeutics.

Marine invertebrate natural products for anti-inflammatory and chronic diseases.

The marine environment represents a relatively available source of functional ingredients that can be applied to various aspects of food processing, storage, and fortification. Moreover, numerous marine invertebrates based compounds have biological activities and also interfere with the pathogenesis of diseases. Isolated compounds from marine invertebrates have been shown to pharmacological activities and are helpful for the invention and discovery of bioactive compounds, primarily for deadly diseases like cancer, acquired immunodeficiency syndrome (AIDS), osteoporosis, and so forth. Extensive research within the last decade has revealed that most chronic illnesses such as cancer, neurological diseases, diabetes, and autoimmune diseases exhibit dysregulation of multiple cell signaling pathways that have been linked to inflammation. On the basis of their bioactive properties, this review focuses on the potential use of marine invertebrate derived compounds on anti-inflammatory and some chronic diseases such as cardiovascular disease, osteoporosis, diabetes, HIV, and cancer.

Protective effect of zinc on N-methyl-N-nitrosourea and testosterone-induced prostatic intraepithelial neoplasia in the dorsolateral prostate of Sprague Dawley rats.

Previous studies have suggested that zinc exerts anticarcinogenic and antiproliferative effects against prostate cancer both in vitro and in rat ventral prostate. Zinc accumulation diminishes early in the course of prostate malignancy and it inhibits the growth of several carcinoma cells through induction of cell cycle arrest and apoptosis. In this study, we have investigated the influence of zinc on N-methyl-N-nitrosourea (MNU) and testosterone (T)-induced prostatic intraepithelial neoplasia in the dorsolateral prostate of Sprague Dawley (SD) rats. The results indicate that zinc plays an important role in prostate carcinogenesis. Increased tumor incidence was accompanied by a decrease in prostatic acid phosphatase activity, citrate, zinc, glutathione-S-transferase, reduced glutathione, p53, B-cell lymphoma protein (Bcl-2)-associated X protein and caspase-3 levels in MNU + T-treated rats. On the contrary, significantly increased phase I drug metabolizing enzyme activities, lipid peroxide, hydrogen peroxide, proliferating cell nuclear antigen, Bcl-2 and Bcl-X(L) protein levels were observed in the dorsolateral prostate of MNU + T-treated rats. Simultaneous zinc supplementation significantly reversed these effects in MNU + T-treated rats. Signs of dysplasia, a characteristic of prostatic intraepithelial neoplasia, were evident in the dorsolateral prostatic tissue sections by MNU + T administration. However, zinc supplementation has reversed these effects in the dorsolateral prostatic histoarchitecture. These results suggest that zinc may act as an essential trace element against MNU and testosterone-induced prostatic preneoplastic progression in SD rats.

Quercetin inhibits invasion, migration and signalling molecules involved in cell survival and proliferation of prostate cancer cell line (PC-3).

Urokinase-type plasminogen activator (uPA) is a serine protease that is involved in cancer progression, especially invasion and metastasis including prostate cancer. uPA activation is mediated by transactivation of uPAR and epidermal growth factor receptor (EGF-R) in prostate cancer progression. Prostate cancer (PC-3) cells have highly invasive capacity and they express uPA and uPAR gene. PC-3 cells are treated with quercetin, which inhibits invasion and migration of PC-3 cells. Quercetin downregulates uPA, uPAR and EGF, EGF-R mRNA expressions. Quercetin inhibits cell survival factor ß-catenin, NF-κB and also proliferative signalling molecules such as p-EGF-R, N-Ras, Raf-1, c.Fos c.Jun and p-c.Jun protein expressions. But quercetin increased p38 mitogen-activated protein kinase protein expression. Our results suggest that quercetin inhibit migration and invasion of prostate cancer cells. It shows the value for treatment of invasive and metastasis type of prostate cancer.

Induction of apoptosis and inhibition of PI3K/Akt pathway in PC-3 and LNCaP prostate cancer cells by ethanolic neem leaf extract.

AIM OF THE STUDY: The present study is aimed to delineate the effect of ethanolic neem leaf extract on PI3K/Akt and apoptotic pathway in prostate cancer cell lines (PC-3 and LNCaP). MATERIALS AND METHODS: To test the hypothesis, two different prostate cancer cell lines LNCaP (androgen dependent) and PC-3 (androgen independent) were taken. Cells were exposed to various concentrations of ethanolic neem leaf extract (ENLE) (25-125 µg/ml). The doses were fixed by cell viability (MTT) assay. For apoptotic detection in situ apoptosis assay, caspase-3 activity and protein expression of cytochrome c and Poly-ADP Ribose Polymerase (PARP) were analysed as well as mRNA expression of Bcl-2 family proteins was studied by RT-PCR. The phosphoinositide 3-kinase (PI3K) and p-Akt were analysed by western blotting and mRNA expression of Akt 1 and 2, PTEN was performed by RT-PCR. Immunoblotting of cyclin D1 and p21 was done to access the inhibition of cell proliferation. RESULTS: ENLE gives 50% inhibition at a dose of 100 µg/ml in both PC-3 and LNCaP cells and considered as effective dose. ENLE decreased the protein expression of PI3K as well as p-Akt and the mRNA expression of Akt 1and 2 in both the cells. There was a significant decrease in mRNA expression of PTEN in LNCaP cells. ENLE induced apoptosis and inhibited cell proliferation by inhibiting PI3K/Akt pathway. Decrease in p-Akt leads to increase in the protein level of Bad, p21 and decrease in the cyclin D1, respectively. ENLE treatment increased the cytochrome c expression and caspase-3 activity as well as regulated the mRNA expression of Bcl-2 family proteins thereby inducing apoptosis to both the cell lines. In situ apoptosis assay showed increased red fluorescence in 100 µg/ml of ENLE in both PC-3 and LNCaP cell lines. CONCLUSIONS: The results suggested that ENLE induces apoptosis and inhibits cell proliferation through inhibiting PI3K/Akt pathway in both PC-3 and LNCaP cells.