Objective assessment of stored blood quality by deep learning.

Stored red blood cells (RBCs) are needed for life-saving blood transfusions, but they undergo continuous degradation. RBC storage lesions are often assessed by microscopic examination or biochemical and biophysical assays, which are complex, time-consuming, and destructive to fragile cells. Here we demonstrate the use of label-free imaging flow cytometry and deep learning to characterize RBC lesions. Using brightfield images, a trained neural network achieved 76.7% agreement with experts in classifying seven clinically relevant RBC morphologies associated with storage lesions, comparable to 82.5% agreement between different experts. Given that human observation and classification may not optimally discern RBC quality, we went further and eliminated subjective human annotation in the training step by training a weakly supervised neural network using only storage duration times. The feature space extracted by this network revealed a chronological progression of morphological changes that better predicted blood quality, as measured by physiological hemolytic assay readouts, than the conventional expert-assessed morphology classification system. With further training and clinical testing across multiple sites, protocols, and instruments, deep learning and label-free imaging flow cytometry might be used to routinely and objectively assess RBC storage lesions. This would automate a complex protocol, minimize laboratory sample handling and preparation, and reduce the impact of procedural errors and discrepancies between facilities and blood donors. The chronology-based machine-learning approach may also improve upon humans' assessment of morphological changes in other biomedically important progressions, such as differentiation and metastasis.

Nano-technological interventions in crop production-a review.

Agricultural industry is facing huge crisis due to fast changing climate, decreased soil fertility, macro and micronutrient insufficiency, misuse of chemical fertilizers and pesticides, and heavy metal presence in soil. With exponential increase in world's population, food consumption has increased significantly. Maintaining the production to consumption ratio is a significant challenge due to shortage caused by various issues faced by agricultural industry even with the improved agricultural practices. Recent scientific evidence suggests that nanotechnology can positively impact the agriculture sector by reducing the harmful effects of farming operations on human health and nature, as well as improving food productivity and security. Farmers are combining improved agricultural practices like usage of fertilizers, pesticides etc. with nano-based materials to improve the efficiency and productivity of crops. Nano technology is also playing a significant role improving animal health products, food packaging materials, and nanosensors for detecting pathogens, toxins, and heavy metals in soil among others. The nanobased materials have improved the productivity twice with half the resources being utilized. Nanoparticles that are currently in use include titanium dioxide, zinc oxide, silicon oxide, magnesium oxide, gold, and silver used for increasing soil fertility and plant growth. Crop growth, yield, and productivity are improved by controlled release nanofertilizers. In this review we elaborate on the recent developments in the agricultural sector by the usage of nanomaterial based composites which has significantly improved the agricultural sector especially how nanoparticles play an important role in plant growth and soil fertility, in controlling plant diseases by the use of nanopesticides, nanoinsecticides, nanofertilizers, Nanoherbicides, nanobionics, nanobiosensors. The review also highlights the mechanism of migration of nanoparticles in plants and most importantly the effects of nanoparticles in causing plant and soil toxicity.

Unlocking the potential of biosynthesized zinc oxide nanoparticles for degradation of synthetic organic dyes as wastewater pollutants.

The azo dyes released into water from different industries are accumulating in the water bodies and bioaccumulating within living systems thereby affecting environmental health. This is a major concern in developing countries where stringent regulations are not followed for the discharge of industrial waste into water bodies. This has led to the accumulation of various pollutants including dyes. As these developing countries also face acute water shortages and due to the lack of cost-effective systems to remove these pollutants, it is essential to remove these toxic dyes from water bodies, eradicate dyes, or generate fewer toxic derivatives. The photocatalysis mechanism of degradation of azo dyes has gained importance due to its eco-friendly and non-toxic roles in the environment. The zinc nanoparticles act as photocatalysts in combination with plant extracts. Plant-based nanoparticles over the years have shown the potential to degrade dyes efficiently. This is carried out by adjusting the dye and nanoparticle concentrations and combinations of nanoparticles. Our review article considers increasing the efficiency of degradation of dyes using zinc oxide (ZnO) nanoparticles and understanding the photocatalytic mechanisms in the degradation of dyes and the toxic effects of these dyes and nanoparticles in different tropic levels.

Human Immunodeficiency Virus Type 1 Phylodynamics to Detect and Characterize Active Transmission Clusters in North Carolina.

BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) phylodynamics can be used to monitor epidemic trends and help target prevention through identification and characterization of transmission clusters. METHODS: We analyzed HIV-1 pol sequences sampled in North Carolina from 1997 to 2014. Putative clusters were identified using maximum-likelihood trees and dated using Bayesian Markov Chain Monte Carlo inference. Active clusters were defined as clusters including internal nodes from 2009 to 2014. Effective reproductive numbers (Re) were estimated using birth-death models for large clusters that expanded ≥2-fold from 2009 to 2014. RESULTS: Of 14 921 persons, 7508 (50%) sequences were identified in 2264 clusters. Only 288 (13%) clusters were active from 2009 to 2014; 37 were large (10-36 members). Compared to smaller clusters, large clusters were increasingly populated by men and younger persons; however, nearly 60% included ≥1 women. Clusters with ≥3 members demonstrated assortative mixing by sex, age, and sample region. Of 15 large clusters with ≥2-fold expansion, nearly all had Re approximately 1 by 2014. CONCLUSIONS: Phylodynamics revealed transmission cluster expansion in this densely sampled region and allowed estimates of Re to monitor active clusters, showing the propensity for steady, onward propagation. Associations with clustering and cluster characteristics vary by cluster size. Harnessing sequence-derived epidemiologic parameters within routine surveillance could allow refined monitoring of local subepidemics.

Factors Associated With Human Immunodeficiency Virus Infections Linked in Genetic Clusters But Disconnected in Partner Tracing.

BACKGROUND: Successful partner notification can improve community-level outcomes by increasing the proportion of persons living with human immunodeficiency virus (HIV) who are linked to HIV care and virally suppressed, but it is resource intensive. Understanding where HIV transmission pathways may be undetected by routine partner notification may help improve case finding strategies. METHODS: We combined partner notification interview and HIV sequence data for persons diagnosed with HIV in Wake County, NC in 2012 to 2013 to evaluate partner contact networks among persons with HIV pol gene sequences 2% or less pairwise genetic distance. We applied a set of multivariable generalized estimating equations to identify correlates of disparate membership in genetic versus partner contact networks. RESULTS: In the multivariable model, being in a male-male pair (adjusted odds ratio [AOR], 16.7; P = 0.01), chronic HIV infection status (AOR, 4.5; P < 0.01), and increasing percent genetic distance between each dyad member's HIV pol gene sequence (AOR, 8.3 per each 1% increase, P < 0.01) were all associated with persons with HIV clustering but not being identified in the partner notification network component. Having anonymous partners or other factors typically associated with risk behavior were not associated. CONCLUSIONS: Based on genetic networks, partnerships which may be stigmatized, may have occurred farther back in time or may have an intervening partner were more likely to be unobserved in the partner contact network. The HIV genetic cluster information contributes to public health understanding of HIV transmission networks in these settings where partner identifying information is not available.

Emerging use of machine learning and advanced technologies to assess red cell quality.

Improving blood product quality and patient outcomes is an accepted goal in transfusion medicine research. Thus, there is an urgent need to understand the potential adverse effects on red blood cells (RBCs) during pre-transfusion storage. Current assessment techniques of these degradation events, termed "storage lesions", are subjective, labor-intensive, and complex. Here we describe emerging technologies that assess the biochemical, biophysical, and morphological characteristics of RBC storage lesions. Of these emerging techniques, machine learning (ML) has shown potential to overcome the limitations of conventional RBC assessment methods. Our previous work has shown that neural networks can extract chronological progressions of morphological changes in RBCs during storage without human input. We hypothesize that, with broader training and testing of multivariate data (e.g., varying donor factors and manufacturing methods), ML can further our understanding of clinical transfusion outcomes in multiple patient groups.

Botulinum Toxin Type-A for the Treatment of Excessive Gingival Display on Smiling.

AIM AND OBJECTIVE: To determine the effect of botulinum toxin type-A (BTX-A) in reducing gummy smile. MATERIALS AND METHODS: This was an interventional case series conducted at a tertiary care setting in Mangaluru, Karnataka, India. Twenty patients with the gingival display of >3 mm were selected through convenience sampling technique. 2.5 units of BTX-A were injected under sterile conditions on either side of the face. The effect of the injection on gingival display was measured at the interval of 2, 4, 12, and 24 weeks. At the end of the treatment, the patient was asked to smile and was photographed and was compared to pretreatment photo. Repeated measures analysis of variance (ANOVA) was used to detect the change at each interval and intergroup comparisons were performed using post hoc Bonferroni test. RESULTS: The mean gingival exposure reduction from 4.93 to 3.705 mm at week 2. At the end of 24 weeks, the evaluation showed the mean gingival display of 4.755 mm on administration of 2.5 units on either side. The photographs also showed the change in smile at the end of the treatment. CONCLUSION: From the study, it was inferred that botulinum toxin was effective in eliminating gummy smile along with noticeable decrease in the nasolabial fold despite for the effect lasting only about 24 weeks. CLINICAL SIGNIFICANCE: A nonsurgical method of correction of gummy smiles when compared to surgical intervention is readily accepted by patients. Botulinum toxin type-A for gummy smiles is an effective, conservative, and satisfactory technique.

Label-Free Analysis of Red Blood Cell Storage Lesions Using Imaging Flow Cytometry.

Deleterious changes, collectively termed as storage lesions, alter the characteristics of red blood cell (RBC) morphology during in vitro storage. Due to gradual loss of cellular membrane, RBCs lose their original biconcave disk shape and begin a process of spherical deformation that is characterized by well-defined morphological criteria. At the spheroechinocyte stage, the structure of RBC is irreversibly damaged and lacks the elasticity necessary to efficiently deliver oxygen. Quantifying the prevalence of spheroechinocytes could provide an important morphological measure of the quality of stored blood products. Unlike the conventional RBC morphology characterization assay involving light microscopy, we introduce a label-free assay using imaging flow cytometry (IFC). The technique captures 100,000 images of a sample and calculates a relative measure of spheroechinocyte population in a fraction of the time required by the conventional method. A comparative method study, measuring a morphological index for 11 RCC units through storage, found that the two techniques measured similar trends with IFC reporting the metric at an average of 3.9% higher. We monitored 18 RCC units between Weeks 1 and 6 of storage and found that the spheroechinocyte population increased by an average of 26.2%. The large (3.5-64.1%) variation between the units' spheroechinocyte population percentage at Week 1 suggests a possible dependence of blood product quality on donor characteristics. Given our method's ability to rapidly monitor large samples and refine morphological characterization beyond conventional methods, we believe our technique offers good potential for studying the underlying relationships between RBC morphology and blood storage lesions. © 2019 International Society for Advancement of Cytometry.

Bioproduction of fumaric acid: an insight into microbial strain improvement strategies.

Fumaric acid (FA), a metabolic intermediate, has been identified as an important carbohydrate derived platform chemical. Currently, it is commercially sourced from petrochemicals by chemical conversion. The shift to biochemical synthesis has become essential for sustainable development and for the transition to a biobased economy from a petroleum-based economy. The main limitation is that the concentrations of FA achieved during bioproduction are lower than that from a chemical process. Moreover, the high cost associated with bioproduction necessitates a higher yield to improve the feasibility of the process. To this effect, genetic modification of microorganism can be considered as an important tool to improve FA yield. This review discusses various genetic modifications strategies that have been studied in order to improve FA production. These strategies include the development of recombinant strains of Rhizopus oryzae, Escherichia coli, Saccharomyces cerevisiae, and Torulopsis glabrata as well as their mutants. The transformed strains were able to accumulate fumaric acid at a higher concentration than the corresponding wild strains but the fumaric acid titers obtained were lower than that reported with native fumaric acid producing R. oryzae strains. Moreover, one plausible adoption of gene editing tools, such as Agrobacterium-mediated transformation (AMT), CRISPR CAS-9 and RNA interference (RNAi) mediated knockout and silencing, have been proposed in order to improve fumaric acid yield. Additionally, the introduction of the glyoxylate pathway in R. oryzae to improve fumaric acid yield as well as the biosynthesis of fumarate esters have been proposed to improve the economic feasibility of the bioprocess. The adoption of some of these genetic engineering strategies may be essential to enable the development of a feasible bioproduction process.

Feasibility of photoacoustic imaging for the non-invasive quality management of stored blood bags.

BACKGROUND AND OBJECTIVES: During the in vitro storage of red blood cells (RBCs), unfavourable changes (storage lesions) cause a rapid consumption of intracellular diphosphoglycerate. The latter deregulates the oxygen-haemoglobin binding potential, subsequently increasing oxygen saturation (SO2 ) and membrane degradation, transforming RBCs from biconcave discs to rigid spherical bodies (spheroechinocytes). Current laboratory techniques invasively extract RBC samples to assess the quality of red cell concentrate (RCC) units. Optical technologies could provide a means of assessing quality non-invasively. MATERIALS AND METHODS: A photoacoustic (PA) imaging technique was developed for acquiring the SO2 of blood bags non-invasively. Seven RCC units were monitored every 3-5 days until expiry (6 weeks). Measurements were validated against a conventional blood gas analyzer (BGA). Using an image flow cytometry assay, morphological profile trends were compared against the SO2 trends during blood bag storage. RESULTS: A strong correlation (r2  ≥ 0·95) was found when comparing temporal data between PA and BGA SO2 measurements. Inter-sample PA variability was found to be similar to that produced by BGA (±0·8%). A strong correlation was found to exist between the temporal changes in SO2 and relative spheroechinocyte population (0·79 ≤ r2  ≤ 0·97). CONCLUSION: This study suggests that PA imaging can non-invasively track the SO2 of stored RBCs non-invasively. By longitudinally monitoring the change in SO2 , it is possible to infer the effects of the storage lesion on RBC morphology. This non-invasive monitoring technique allows for the assessment of blood bags, without compromising sterility pre-transfusion.

Integration of Contact Tracing and Phylogenetics in an Investigation of Acute HIV Infection.

BACKGROUND: The integration of traditional contact tracing with HIV sequence analyses offers opportunities to mitigate some of the barriers to effective network construction. We used combined analyses during an outbreak investigation of spatiotemporally clustered acute HIV infections to evaluate if the observed clustering was the product of a single outbreak. METHODS: We investigated acute and recent HIV index cases reported in North Carolina from 2013 to 2014 and their reported contacts. Contact tracing networks were constructed with surveillance data and compared with phylogenetic transmission clusters involving an index case using available HIV-1 pol sequences including 1672 references. Clusters were defined as clades of 2 or more sequences with a less than 1.5% genetic distance and a bootstrap of at least 98% on maximum-likelihood phylogenies. RESULTS: In total, 68 index cases and 210 contacts (71 HIV infected) were reported. The contact tracing network involved 58 components with low overall density (1.2% statewide); 33% of first-degree contacts could not be located. Among 38 (56%) of 68 index cases and 34 (48%) of 71 contacts with sequences, 13 phylogenetic clusters were identified (size 2-4 members). Four clusters connected network components that were not linked in contact tracing. The largest component (n = 28 cases) included 2 distinct phylogenetic clusters and spanned 2 regions. CONCLUSIONS: We identified the concurrent expansion of multiple small transmission clusters rather than a single outbreak in a largely disconnected contact tracing network. Integration of phylogenetic analyses provided timely information on transmission networks during the investigation. Our findings highlight the potential of combined methods to better identify high-risk networks for intervention.

Efficient in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood) and assessment of genetic fidelity of micropropagated plants.

A reliable protocol has been established for in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood), a valuable medicinal plant from India. A highly proliferating organogenic callus was obtained on Murashige and Skoog (MS) medium supplemented with 2.5 µM IAA when nodal explants were cultured on MS medium supplemented with various growth regulators. Further, highest regeneration frequency (83.3 %) of adventitious shoots was observed, when the callus was sub-cultured on MS medium supplemented with 6-benzylaminopurine (BAP; 2.5 µM) along with 7.5 µM 2-isopentenyl adenine (2-iP). An optimal of 10.16 ± 2.24 shoots were regenerated on medium supplemented with 2.5 µM BAP + 7.5 µM 2-iP. Quarter strength MS medium supplemented with 10 µM IBA was effective for rooting of the shoots. Ex-vitro plants were normal and were established successfully. Cytological and molecular marker studies showed that regenerated plants showed genetic stability in micro-propagated plants.

An intraoral epitheloid hemangioendothelioma masquerading clinically as pyogenic granuloma.

Epitheloid hemangioendothelioma (EHE) is an uncommon angiocentric neoplasm of intermediate malignant potential. This tumor is frequently seen in the lungs and liver, but its presentation in the oral cavity is rare. In the oral cavity, gingival region is the common sites of occurrence. We report a rare case of epitheloid hemangioendothelioma in a 48-year-old male, presenting as a growth in the upper anterior gingiva of five months duration along with a review of its clinicopathological and immunohistochemical characteristics.

Resistance to HIV integrase strand transfer inhibitors among clinical specimens in the United States, 2009-2012.

BACKGROUND: Data on integrase inhibitor resistance come primarily from clinical trials and in vitro studies. We examined results of all clinically indicated integrase genotypic resistance tests (GRTs) performed at a US national referral lab from 2009 through 2012. METHODS: Integrase sequences and demographic data were compiled with paired protease-reverse transcriptase (PR-RT) GRT results, when available. Analyses utilized the Stanford HIV Drug Resistance Database. "Major" integrase mutations included T66AIK, E92QV, F121Y, Y143CHR, S147G, Q148HKR, and N155H; multiple accessory mutations were also assessed. RESULTS: Among 3294 sequences from 3012 patients, 471 patients had viruses with ≥ 1 raltegravir or elvitegravir resistance mutation (15.6%). Q148 and N155 pathways were equally represented (both n = 197); 84 had Y143 mutations. Q148 rarely occurred without accessory mutations (n = 3). Among 224 patients with serial integrase GRTs, 22 with baseline wild-type acquired a major mutation, after a median 224 days between tests (interquartile range, 148-335 days). Major mutations were observed to persist up to 462 days. Most (62%) had paired PR-RT results. Patients with integrase-resistant viruses were older and more likely to have PR-RT mutations (both P < .001). Among those with PR-RT data, 42 patients had 4-class resistance (2.3%). Sex, geographic region, and test year were not associated with integrase resistance. High-level dolutegravir resistance was predicted in 12% of patients with raltegravir- or elvitegravir-resistant viruses (2% of all patients). CONCLUSIONS: Approximately 1 in 6 US patients undergoing integrase GRT for clinical decision making harbors significant resistance, with Q148 and N155 pathways equally common. Dolutegravir is likely to have full or partial activity against most variants observed.

From Psoriatic Arthritis to Takayasu Arteritis and Stroke: A Challenging Journey of Overlapping Autoimmune Rheumatic Diseases in a Young Female.

This case report details the clinical presentation, diagnosis, and management of a 23-year-old female with a unique medical history, including psoriatic arthritis (PsA) and Takayasu arteritis (TAK) and ultimately presenting with acute ischemic young stroke. The patient initially presented with right hip and buttocks pain, multiple itchy skin lesions, and right sacroiliac joint pain in 2017. She was diagnosed with psoriatic arthritis and treated effectively, achieving complete remission by 2020. In August 2023, she presented with acute-onset neurological deficits and vascular symptoms leading to a diagnosis of TAK. This case highlights the challenges in diagnosing and managing a complicated case with overlapping autoimmune rheumatic diseases.

A Comparative Evaluation of Surface Hardness and Nanomechanical Properties of Nickel-Titanium Orthodontic Wires: An In Vitro Study.

OBJECTIVE: The study aimed to evaluate the influence of various surface coatings (epoxy, Teflon, and rhodium) on the surface roughness (SR) and nanomechanical characteristics of nickel-titanium (NiTi) archwires. The study compared these coated archwires to uncoated ones from a single manufacturer, which served as a control. MATERIALS AND METHODS: There were 15 rectangular samples of four distinct archwires measuring 0.17 × 0.25. These were ultrasonically treated with an alkaline solution at 60°C for 15 minutes before being rinsed with distilled water to remove precipitates. With an orthodontic soft wire cutter, the straight buccal sections of coated and uncoated archwires were cut into 20 mm lengths. A three-dimensional optical noncontact surface profilometer evaluated the surface. Profilometers use contact scanning white light interferometry. Using the Vision64 software (Bruker Corporation, San Jose, CA), the profilometer's nanolens atomic force microscopy module has a completely automated turret with programmed X, Y, and Z motions. Images were taken in five random locations. Five average measurements matched specimen SR. A nanoindenter with a Berkovich diamond indenter measured nanohardness (NH) and elastic modulus (EM). The experimental results were analyzed using the Statistical Package for the Social Sciences software version 26.0 (SPSS Inc., Chicago, IL). To examine mean differences at 5% significance, analysis of variance and Tukey's post hoc test were applied for SR, NH, and EM. RESULTS: Wires coated with epoxy had the highest SR (1.499 ± 0.082), followed by Teflon (0.811 ± 0.023) and rhodium (0.308 ± 0.024). The SR of the control group was 0.289 ± 0.027. Significant differences in SR were found (p < 0.0001). Except for the comparison between rhodium and the control group (p = 0.684), all intergroup comparisons of SR showed significant differences (p < 0.0001). The rhodium-coated wires exhibited the highest NH (0.185 ± 0.014), and the epoxy group had the lowest (0.147 ± 0.017). Variations in NH were significant between the study groups (p < 0.0001). The epoxy, Teflon, and rhodium groups showed significant differences against the control group (p < 0.0001) in intergroup comparisons for NH. The Teflon group had the highest EM (5.367 ± 0.379), and the epoxy group had the lowest (5.012 ± 0.498). The EM of the control group was 56.946 ± 0.737. Results indicate considerable EM changes between the groups (p < 0.05). Comparisons between experimental and control groups showed significant differences (p < 0.0001). CONCLUSION: The study's findings indicate that the SR of rhodium-coated archwires is substantially comparable to that of uncoated archwires. However, Teflon-, rhodium-, and epoxy-coated archwires had significantly different NH and EM compared to uncoated ones. Further, uncoated archwires have higher NH and EM.

CLINICAL PROFILES AND OUTCOMES OF ADMISSIONS FOLLOWING COVID-19 ADMISSIONS DURING THREE WAVES OF THE PANDEMIC: EXPERIENCE OF A TERTIARY HOSPITAL IN THE EASTERN CAPE PROVINCE OF SOUTH AFRICA.

Background: South Africa was the country worst affected by the Covid-19 pandemic in Africa. There is a paucity of data on the clinical characteristics and mortality of Covid-19 from the Eastern Cape province of South Africa. We report on the demographic and clinical characteristics as well as the mortality of patients admitted to the Covid-19 ward of Nelson Mandela Academic Hospital (NMAH), Mthatha, during three waves of the Covid-19 pandemic in South Africa. Materials and Methods: We conducted a single centre retrospective observational study of patients admitted for Covid-19 in a tertiary hospital in the rural Eastern Cape of South Africa. Data were collected from patient files, electronic databases and the National Health Laboratory Services (NHLS) database. The outcomes were duration of admission and in-hospital mortality. Results: There were 371 patients admitted across all three waves with a mean age of 52.2 ± 16.3 years. The proportion of females across the three waves is 61.2%. The commonly associated comorbidities, irrespective of the wave, were hypertension, diabetes and HIV infection. The median duration of admission was six days, with an overall mortality of 31%. The mortality for first, second and third wave were 29.3%, 31.5% and 37.9% respectively. Conclusion: Admissions for Covid-19 were predominantly in females and middle-aged. One third of the admitted patients died. Diabetes, hypertension and HIV infection were the most commonly associated comorbidities.

Conservation of Endangered Cordyceps sinensis Through Artificial Cultivation Strategies of C. militaris, an Alternate.

Cordyceps, an entomopathogenic fungus belonging to the Ascomycota phylum, is a familiar remedial mushroom that is extensively used in the traditional medicinal system, especially in South Asian nations. The significance of this genus' members in a range of therapeutic and biotechnological applications has long been acknowledged. The exceedingly valuable fungus Ophiocordyceps sinensis (Cordyceps sinensis) is found in the alpine meadows of Bhutan, Nepal, Tibet, and India, where it is severely harvested. Driven by market demand and ecological concerns, the study highlights challenges in natural C. sinensis collection and emphasizes the shift towards sustainable artificial cultivation methods. This in-depth review navigates Cordyceps cultivation strategies, focusing on C. sinensis and the viable alternative, C. militaris. The escalating demand for Cordyceps fruiting bodies and bioactive compounds prompts a shift toward sustainable artificial cultivation. While solid-state fermentation on brown rice remains a traditional method, liquid culture, especially submerged and surface/static techniques, emerges as a key industrial approach, offering shorter cultivation periods and enhanced cordycepin production. The review accentuates the adaptability and scalability of liquid culture, providing valuable insights for large-scale Cordyceps production. The future prospects of Cordyceps cultivation require a holistic approach, combining scientific understanding, technological innovation, and sustainable practices to meet the demand for bioactive metabolites while ensuring the conservation of natural Cordyceps populations.