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Cytokines are critical mediators that oversee and regulate immune and inflammatory responses via complex networks and serve as biomarkers for many diseases. Quantification of cytokines has significant value in both clinical medicine and biology as the levels provide insights into physiological and pathological processes and can be used to aid diagnosis and treatment. Cytokines and their clinical significance are introduced from the perspective of their pro‐ and anti‐inflammatory effects. Factors affecting cytokines quantification in biological fluids, native levels in different body fluids, sample processing and storage conditions, sensitivity to freeze‐thaw, and soluble cytokine receptors are discussed. In addition, recent advances in in vitro and in vivo assays, biosensors based on different signal outputs and intracellular to extracellular protein expression are summarized. Various quantification platforms for high‐sensitivity and reliable measurement of cytokines in different scenarios are discussed, and commercially available cytokine assays are compared. A discussion of challenges in the development and advancement of technologies for cytokine quantification that aim to achieve real‐time multiplex cytokine analysis for point‐of‐care situations applicable for both biomedical research and clinical practice are discussed. Cytokines are important cellular signaling molecules and immune system mediators. Abnormal cytokine levels may cause cytokine storm and diseases. Consequently, quantification of cytokines is valuable for diseases diagnosisand therapy. The clinical significance of cytokines, factors affecting cytokine quantification, and advances of cytokine detection are summarized, providing a prospective for real‐time quantification of multiplex cytokines in the clinic.

Although drugs currently used for the various types of diseases (e.g., antiparasitic, antiviral, antibacterial, etc.) are effective, they present several undesirable pharmacological and pharmaceutical properties. Most of the drugs have low bioavailability, lack of sensitivity, and do not target only the damaged cells, thus also affecting normal cells. Moreover, there is the risk of developing resistance against drugs upon chronic treatment. Consequently, their potential clinical applications might be limited and therefore, it is mandatory to find strategies that improve those properties of therapeutic agents. The development of prodrugs using amino acids as moieties has resulted in improvements in several properties, namely increased bioavailability, decreased toxicity of the parent drug, accurate delivery to target tissues or organs, and prevention of fast metabolism. Herein, we provide an overview of models currently in use of prodrug design with amino acids. Furthermore, we review the challenges related to the permeability of poorly absorbed drugs and transport and deliver on target organs.

Seaweed produces metabolites aiding in the protection against different environmental stresses. These compounds show antiviral, antiprotozoal, antifungal, and antibacterial properties. Macroalgae can be cultured in high volumes and would represent an attractive source of potential compounds useful for unconventional drugs able to control new diseases or multiresistant strains of pathogenic microorganisms. The substances isolated from green, brown and red algae showing potent antimicrobial activity belong to polysaccharides, fatty acids, phlorotannins, pigments, lectins, alkaloids, terpenoids and halogenated compounds. This review presents the major compounds found in macroalga showing antimicrobial activities and their most promising applications.

Leishmaniasis is a neglected tropical disease caused by protozoan parasites of the genus Leishmania. An estimated 700,000 to 1 million new cases occur annually. Current therapies are limited by high toxicity, cost, prolonged treatment period, and rising resistance in endemic regions. The Asteraceae family has emerged as a promising source of bioactive compounds with proven leishmanicidal activity. In this study, the assessment of the antileishmanial activity of Helianthus tuberosus and Vernonanthura squamulosa extracts, the isolation of the sesquiterpene lactones heliangin and glaucolide A, respectively, and the evaluation of the activity of the compounds were conducted. Dichloromethane extracts of H. tuberosus and V. squamulosa were active on Leishmania amazonensis promastigotes, inhibiting the replication of the parasites in 97.2 ± 3.1% and 89.1 ± 1.1%, respectively, at 100 μg/mL. Heliangin was active against promastigotes of L. amazonensis (IC50 = 9.3 μM) and intracellular amastigotes (IC50 = 0.8 μM), while glaucolide A exhibited moderate activity against promastigotes (IC50 = 46.7 μM) and did not show activity against intracellular amastigotes. Based on these results, heliangin was further evaluated in an animal model of cutaneous leishmaniasis using BALB/c mice infected with L. amazonensis. Heliangin (8 mg/Kg), when administered in combination with Glucantime, significantly reduced lesion progression and parasite load compared to the vehicle-treated group (p < 0.001). These findings show that heliangin is a potential candidate for leishmaniasis treatment, especially in combination with therapeutic drugs.

This study reports on the pharmacological characterization of ZP120, a novel ligand of the nociceptin/orphanin FQ (N/OFQ) peptide receptor, NOP. ZP120 is a structure inducing probes modified NOP ligand: Zealand Pharma proprietary SIP technology was used to increase the enzymatic stability and half‐life of peptide. In vitro, ZP120 mimicked the inhibitory effects of N/OFQ in the electrically stimulated mouse vas deferens, showing however higher potency (pEC50 8.88 vs 7.74), lower maximal effects (Emax 69±5% vs 91±2%), and slower onset of action. Like N/OFQ, the effects of ZP120 were not modified by 1 μM naloxone, but they were antagonized by the NOP receptor selective antagonist J‐113397 (pA2 7.80 vs ZP120, 7.81 vs N/OFQ). In vivo, ZP120 mimicked the effects of N/OFQ, producing pronociceptive effects in the tail withdrawal assay and decreased locomotor activity after i.c.v., but not after i.v. administration in mice. ZP120 elicited similar maximal effects as N/OFQ, but it was about 10 fold more potent and its effects lasted longer. In conclusion, the novel NOP receptor ligand ZP120 is a highly potent and selective partial agonist of the NOP receptor with prolonged effects in vivo. British Journal of Pharmacology (2002) 137, 369–374. doi:10.1038/sj.bjp.0704894

Available treatments for hepatitis B can control the virus but are rarely curative. This led to a global initiative to design new curative therapies for the 257 million patients affected. Discovery and development of these new therapies is contingent upon functional in vitro and in vivo hepatitis B virus (HBV) infection models. However, low titer and impurity of conventional HBV stocks reduce significance of in vitro infections and moreover limit challenge doses in current in vivo models. Therefore, there is a critical need for a robust, simple and reproducible protocol to generate high-purity and high-titer infectious HBV stocks. Here, we outline a three-step protocol for continuous production of high-quality HBV stocks from supernatants of HBV-replicating cell lines. This purification process takes less than 6 h, yields to high-titer stocks (up to 1 × 1011 enveloped, DNA-containing HBV particles/mL each week), and is with minimal equipment easily adaptable to most laboratory settings.

Finding new, safe strategies to prevent and control rheumatoid arthritis is an urgent task. Bioactive peptides and peptide‐rich protein hydrolyzate represent a new trend in the development of functional foods and nutraceuticals. The resulting tissue hydrolyzate of the chicken embryo (CETH) has been evaluated for acute toxicity and tested against chronic arthritis induced by Freund's full adjuvant (modified Mycobacterium butyricum) in rats. The antiarthritic effect of CETH was studied on the 28th day of the experiment after 2 weeks of oral administration of CETH at doses of 60 and 120 mg/kg body weight. Arthritis was evaluated on the last day of the experiment on the injected animal paw using X‐ray computerized microtomography and histopathology analysis methods. The CETH effect was compared with the non‐steroidal anti‐inflammatory drug diclofenac sodium (5 mg/kg). Oral administration of CETH was accompanied by effective dose‐dependent correction of morphological changes caused by the adjuvant injection. CETH had relatively high recovery effects in terms of parameters for reducing inflammation, inhibition of osteolysis, reduction in the inflammatory reaction of periarticular tissues, and cartilage degeneration. This study presents for the first time that CETH may be a powerful potential nutraceutical agent or bioactive component in the treatment of rheumatoid arthritis. Finding new, safe strategies to prevent and control rheumatoid arthritis is an urgent task. Bioactive peptides and peptide‐rich protein hydrolyzate represent a new trend in the development of functional foods and nutraceuticals. This study presents for the first time that CETH may be a powerful potential nutraceutical agent or bioactive component in the treatment of rheumatoid arthritis.

To determine Fusarium species and their pathogenicity in maize-production areas of the Tepebaşı, Odunpazarı, Alpu, and Seyitgazi districts of Eskişehir province, Türkiye, 180 samples were collected from 45 fields during survey studies conducted in 2023–2024. A total of 110 Fusarium isolates were obtained from the collected plant samples. The isolates were identified as F. verticillioides, F. culmorum, F. proliferatum, F. graminearum, F. sambucinum, F. acuminatum, F. chlamydosporum, and F. equiseti. The most common species was F. verticillioides, while the most virulent species was F. graminearum, with a disease severity of 96.67%. The effects of different doses of soil-applied herbicides containing the active ingredients Isoxaflutole + Thiencarbazone-methyl + Cyprosulfamide, Dimethenamid-P + Saflufenacil, and S-Metolachlor + Terbuthylazine on F. graminearum were evaluated under both in vitro and in vivo conditions. Under in vitro conditions, the highest inhibition rate (57.23%) was observed in the double-dose application of the herbicide containing S-Metolachlor + Terbuthylazine. This was followed by the upper and recommended doses of the same herbicide with inhibition rates of 47.16% and 39.46%, respectively. For the other herbicides, inhibition rates increased with increasing herbicide dose. In field trials, the highest suppression of the pathogen was also observed with the herbicide containing S-Metolachlor + Terbuthylazine. While the recommended dose showed a 38.6% effect against the pathogen, the upper dose resulted in a 45.31% effect. This study suggests that herbicide applications may be associated with improved plant growth, likely due to reduced pathogen pressure and decreased weed competition. The findings highlight the complex interactions between soil-applied herbicides, soil-borne pathogens, and host plants, and provide insights into the development of integrated disease management strategies in maize-production systems.

Based on a publication of Tomasetti and Vogelstein in 2015, in which the risk of cancer development is postulated to be just one-third caused by genetic predisposition and environmental factors, it seemed worth focusing again on the value of test systems for screening chemicals for their carcinogenicity. This review aims to firstly summarize data on a host-mediated in vivo/in vitro assay developed by our working group to screen the tumorigenic potential of chemicals. Subsequently, in this article the importance and advantages of host-mediated in vivo/in vitro assays in general have been compared with in vivo and in vitro tests. The applicability of the host-mediated in vivo/in vitro assay system within broad screening strategies is discussed. The main intention of this review is to stimulate developments of newer approaches in the field of carcinogenic testing.

The efficacy of different fungicides, belonging to different chemical groups, was in vitro and in vivo tested in glasshouse trials against Allophoma tropica, the causal agent of a new leaf spot on lettuce. The in vitro trials showed that only a few fungicides were effective in inhibiting the mycelial growth of the four tested strains of A. tropica: the most effective fungicide, with the lowest ED₅₀ and MIC values, was penconazole. Prochloraz, propiconazole, boscalid and fludioxonil showed low ED₅₀ values, but high MIC values. The results from the in vivo trials showed that all the tested fungicides significantly reduced Phoma leaf spot incidence and severity, although differences were observed in the achieved disease pressure in the four trials. Generally, in the presence of an average disease severity in the untreated control, the best disease severity reduction (up to 90%) was provided 9 days after the application of fludioxonil, mancozeb, pyraclostrobin + boscalid, propiconazole and iprodione. Fludioxonil and iprodione showed a persistent effect (up to 90%) still being evident 12 days after the application. Generally, in the presence of a high disease severity in the untreated control, 9 days after the application of fludioxonil, mancozeb, iprodione and pyraclostrobin + boscalid provided a disease severity reduction up to 80%. Moreover, copper provided a disease severity reduction from 84 to 89%, applied as preventative treatment, resulting of special interest, particularly for crops grown under organic farming rules.