Nova Biologica Reperta

Abstract Objective: In this study, pure poly lactic acid (PLA) and glutaraldehyde-modified PLA films were prepared using the solvent casting method.
Method: Their structural characteristics, physiological stability, and biocompatibility were evaluated. The structure and properties of the samples were characterized using tensile testing, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). Furthermore, stability in a physiological environment and biocompatibility with Hu02 cells were assessed using the MTT assay.
Results: FTIR and XRD results confirmed the structural modifications. Mechanically, modification with glutaraldehyde led to an increase in tensile strength by approximately 15.26%. Physiological stability analysis revealed that after 28 days, the mass loss percentage was 5.18% for pure PLA films and 5.99% for PLA-glutaraldehyde films. In the biocompatibility assay, the cell viability of the PLA-glutaraldehyde films decreased to below 50% after 5 days.
Conclusions: The results demonstrated that modifying PLA with glutaraldehyde can improve the structural and mechanical properties of the films; however, this improvement is accompanied by potential challenges related to the release of glutaraldehyde residues. Therefore, these materials are potentially suitable for biomedical and biodegradable packaging applications, provided that formulation optimization is conducted to control potential toxicity.

Abstract Objective: The genus Linaria Mill. (Plantaginaceae) represents a taxonomically complex group characterized by high morphological similarity and extensive intraspecific variation, which often complicates the delimitation of species.
Method: In this study, four taxa belonging to the Linaria lineolata s.s. complex (Linaria lineolata, Linaria elymaitica, Linaria karajensis, and Linaria khalkhalensis) were investigated using an integrative approach that combined morphological and molecular data. A total of 20 quantitative and qualitative morphological characters were analyzed using univariate statistics and Principal Component Analysis (PCA). To investigate molecular relationships, sequences of the low-copy nuclear gene AGT1 and the chloroplast intergenic spacer rpl32–trnL were analyzed, and phylogenetic relationships were reconstructed using Bayesian inference.
Results: The PCA results revealed a clear separation among the studied taxa, with the first two principal components explaining a substantial proportion of the total variance. Molecular phylogenetic analyses based on the nuclear gene AGT1 and the chloroplast intergenic spacer rpl32–trnL further supported the delimitation of the studied taxa, revealing distinct and well-supported clades.
Conclusions: A dichotomous identification key was also constructed using the most diagnostic morphological characters. The congruence between morphological and molecular datasets indicates clear taxonomic independence among the investigated species. These findings provide evidence for the delimitation of species boundaries within the L. lineolata s.s. complex and highlight the effectiveness of an integrative taxonomic approach in resolving species complexes within Linaria.

Abstract Clustered regularly interspaced short palindromic repeats (CRISPR) were discovered with the discovery of prokaryotic CRISPR-related proteins called Cas, which are capable of creating double-strand breaks in target sequences in the mammalian genome with the help of guide RNA. This discovery led to the development of CRISPR systems that harness natural DNA repair mechanisms to precisely create mutations in defective genes. CRISPR has been used to remove deleterious mutated genes and correct errors in protein-coding sequences to rescue disease phenotypes in preclinical studies and in several clinical trials. However, most genetic disorders result from a combination of other mutations involving deletions and duplications in both coding and non-coding regions of the genome and therefore require complex genome engineering strategies beyond simple gene deletion. To overcome this limitation, the toolbox of natural and engineered CRISPR-Cas systems has expanded dramatically to include a variety of tools. The use of CRISPR technology to edit non-coding genomes, regulate gene expression, make precise genetic changes, and target infectious diseases has the potential to lead to therapeutic solutions for many incurable diseases.

Abstract Jujube (Ziziphus jujuba Mill.) is a valuable medicinal plant belonging to the family Rhamnaceae. Owing to its rich content of vitamins, pectin, alkaloids, phenolic compounds, and fatty acids, it is of considerable importance in the pharmaceutical industry. Despite the economic and medicinal significance of jujube, information on the diversity of its chemical profile and variations in fruit metabolite composition among different populations in Iran remains limited. In the present study, the chemical profiles of 11 jujube populations collected from eight provinces of Iran were investigated and compared using gas chromatography–mass spectrometry (GC–MS). The obtained data were evaluated using multivariate statistical methods and bioinformatics analyses to determine the patterns of similarity and differentiation among populations based on their chemical composition. The results were evaluated using multivariate statistical and chemometric analyses. Analysis of the GC–MS results revealed significant differences in the chemical composition of fruits among different regions. Based on the similarities and differences in their chemical compositions, the geographical populations were classified into two distinct clusters using Ward's hierarchical clustering method. The results of this study demonstrated that comparing the chemical profiles of different jujube populations can provide valuable information on the metabolic diversity of this species and may contribute to the identification of germplasm resources, the selection of superior populations, and breeding and conservation programs.

Abstract Objective: Dracocephalum kotschyi Boiss. has medicinal properties due to its valuable compounds. The aim of this study was to investigate the effects of red or blue light and sodium nitroprusside (SNP) on enhancing secondary metabolites in D. kotschyi
Method: Seven-day-old seedlings treated under white and red-blue combined spectrum, and were irrigated with Hoagland's solution. After five weeks, half of the plants were harvested, while the others were subjected to SNP treatment at concentrations of 0, 0.5, and 1 µM. The experiments were conducted in three replicates using a completely randomized block design, and data were analyzed using one-way analysis of variance.
Results: Under blue light and in the absence of SNP, the levels of H₂O₂ and MDA of the plants were lower than those in samples grown under white light. Under both white and blue light, SNP application, in a concentration-dependent manner, reduced pigment contents while increasing peroxidase and phenylalanine ammonia-lyase activities. Additionally, accumulation of proline, polysaccharides, phenolic compounds, and flavonoids was observed. Under white light, chlorophyll b content, reducing sugars, polysaccharides, and superoxide dismutase activity increased in the presence of SNP. Red-blue light (at a 1:1 ratio) treatment resulted in increased chlorophyll and carotenoid contents, along with decreased levels of H₂O₂, proline, reducing sugars, polysaccharides, phenolic compounds, flavonoids, and reduced activities of antioxidant enzymes and PAL, even in the presence of SNP.
Conclusions: Cultivation of D. kotschyi under blue light together with SNP enhanced secondary metabolite content by activating the plant's antioxidant defense system, PAL, and modulating carbon metabolism.

Abstract Objective: Helicobacter pylori infection is a major cause of chronic gastritis, peptic ulcer disease, and gastric cancer, highlighting the importance of rapid and reliable diagnostic methods. In the present study, a lateral flow immunoassay (LFIA)-based strip immunobiosensor was developed for the rapid detection of H. pylori using colloidal gold nanoparticles as visual signal reporters.
Method: Colloidal gold nanoparticles (approx. 40 nm) were conjugated with a monoclonal antibody specific to H. pylori to form a nanobioprobe. The conjugation was confirmed using UV–Vis spectroscopy, showing characteristic absorption peaks and indicating stable colloidal behavior after antibody immobilization. The LFIA strip was assembled using a sample pad, conjugate pad, nitrocellulose membrane (with test and control lines), and an absorbent pad. Human serum samples were applied to the strip to evaluate the biosensor’s performance.
Results: The developed diagnostic strip successfully identified positive samples at concentrations as low as 0.02 µg/mL, as indicated by the appearance of distinct colored lines in both the test and control regions. Furthermore, the assay maintained reliable detection performance over different time intervals, indicating acceptable stability of the system.
Conclusions: Overall, the developed LFIA strip represents a simple, rapid, and cost‑effective diagnostic platform for the detection of H. pylori and shows potential for use in point‑of‑care testing.

Abstract Objective: COVID-19, caused by SARS-CoV-2, highlighted the need for the development of therapeutic strategies. Since drug discovery processes are time-consuming and costly, the use of bioinformatics approaches and molecular modeling has gained considerable attention. In this context, therapeutic peptides, due to their high specificity and flexibility, are considered promising candidates for the development of antiviral drugs.
Method: Inhibitory peptides targeting the SARS-CoV-2 main protease (Mpro) were designed and evaluated. Molecular docking was performed using AutoDock, and optimized grid parameters were used in PyRx for virtual screening. Peptides of 2–10 amino acids were generated using unidirectional and bidirectional incremental construction approaches. After structural optimization, peptides were evaluated with a reference ligand and analyzed for binding interactions.
Results: Bidirectionally designed peptides showed more favorable binding energies and better performance than unidirectional ones. Increasing peptide length generally improved binding affinity and complex stability. Decapeptides exhibited the strongest interactions. Several peptides showed lower binding energies than the crystallographic ligand and RMSD values close to zero, indicating stable complexes.
Conclusions: Longer peptides, especially those designed via bidirectional construction, enhance binding to Mpro and improve inhibitory potential. These results provide a rational basis for designing novel peptide-based inhibitors against SARS-CoV-2.

Abstract Finger length ratios are organized during embryonic development of fingers as they exposed to sex steroid hormones, and may show varying degrees of sexual dimorphism between males and females in different animal groups. Among all the finger length ratios calculated in a sample, the ratio between the second to fourth fingers (2D: 4D) is the most important one. In this study, the 2D:4D ratios in both sides of the body (right and left) in all limbs were investigated to determine if sexual dimorphism is present in the 2D: 4D ratios in 44 specimens studied (20 males and 24 females) of Acanthodactylus blanfordi. Other morphological traits of the two sexes were also examined (23 metric and meristic traits), as a result, sexual dimorphism was observed in five metric and meristic morphological traits. In terms of the ratio of the size of the fingers and toes, sexual dimorphism in the length of the fingers was observed only in 2D: 4D on the right side of the body in forelimbs and hindlimbs. The value of this trait was higher in males than females and this difference was statistically significant P ≤0.05).

Abstract Aloe vera L. is one of the most valuable plants in the pharmaceutical, cosmetic, sanitary and food industries. In vitro culture is used for commercial production and due to the abundant application of this plant, extensive research has been performed on the in vitro culture of Aloe vera. For this purpose, the present study was conducted at two stages. At the first stage, the best method of sterilization of explants derived from Aloe vera offshoots was investigated. At the second stage, the effect of the type of explants, the light condition (dark and bright) and the effect of BAP (Benzyl Amino Purine) and NAA (α-Naphthalene acetic acid)) growth regulators on regeneration and the amount of phenolic compounds were studied. A factorial experiment was executed on the basis of a completely randomized design with three replications. The best sterilization protocol was 0.1% mercuric chloride (for 2 minutes), 70% ethanol (for 30 seconds) and 15% sodium hypochlorite (for 5 minutes). The little white explant derived from the base of leaves, with the lowest percentage of phenol and the highest survival rate (67.5%) in darkness, was found to be the best candidate. MS medium supplemented with 0.75 mg / L BAP and 0.25 mg / L NAA resulted in the highest stem number (2.5) and stem length (42.107 mm), establishment percentage (73%), leaf number (6.33), leaf diameter (4.8 mm), chlorophyll b (9.216 mg/g) and carotenoids (4.81 mg/g). The highest content of chlorophyll a (56.07 mg/g) and total chlorophyll (61.35 mg/g) were found in samples treated with hormonal medium, supplemented with 1.5 mg / L of BAP with 0.5 mg / L of NAA. The maximum number (3) and average length (33.3 mm) of roots were observed in samples treated with the hormone-free medium.

Abstract Nerve growth factor (NGF) is a neurotrophic factor that is functional in the survival, maintenance and differentiation of nervous system cells. This protein has three subunits, of which the beta subunit has the main activity. Its structure consists of a cysteine knot motif made up of beta strands linked by disulfide bonds. It can be used as a therapeutic agent in the treatment of many diseases. As NGF extracted from natural sources is unsuitable for therapeutic goals, many studies have attempted to produce recombinant β-NGF. In this study, Trigger Factor (TF) chaperone was expressed simultaneously with β-NGF in E. coli in order to obtain increased yield of soluble recombinant human β-NGF. For this purpose, pET39b (+)::β-NGF and chaperone plasmid pTf16 were transferred to E.coli (DE3 strain). After the induction of each promoter, the total proteins and periplasmic proteins were extracted. To confirm the effects of TF on total protein and soluble β-NGF expression level, Bradford and Dot blot techniques and ImageJ software were used. Then, β-NGF was purified using affinity chromatography column (Ni⁺²-NTA). Also, the PC12 cells were treated with the protein for one week in order to study the function of purified NGF. Our data indicated that co-expression of TF could increase the soluble and periplasmic production of β-NGF but not total proteins. Also, the treatment of PC12 cell line with purified β-NGF, co-expressed with TF chaperone, showed differentiation of these cells to nerve cells. This indicated that the purified NGF is fully functional. Our data suggest that the co-expression of cytoplasmic chaperone (TF) with recombinant nerve growth factor might be an efficient approach to produce a proper quantity of soluble and active rhNGF.

Abstract Introduction: Due to the emergence of antibiotic-resistant strains of bacteria such as Klebsiella pneumoniae, the use of antibacterial potential of bacteriophages has been considered as an effective approach in controlling the infections.
Material and Method: After isolating lytic bacteriophage (PKpMa1/19) against K.pneumoniae PTCC1290 from Maragheh urban sewage by agar bilayer method, purification, titration and enrichment of the bacteriophage were performed. Morphological characteristics of PKpMa1/19 were determined using electron microscopy. One step growth, latent period, burst size, stability and finally host range of the bacteriophage was evaluated using spot test.
Results: A bacteriophage with the morphological features of Tectiviridae family was isolated and purified. Its latent period and burst size were 20 min and 311 PFU respectively. Appropriate lytic effect of the bacteriophage was shown between -22 and 37 . Stability of the phage was high in pH= 4-10. The phage (PKpMa1/19), was effective against one of the 20 clinical isolates of K.pneumoniae (5% of K. pneumoniae isolates), but it showed antibacterial effect on 10 Pseudomonas aeroginosa ( 50% of P. aeruginosa isolates) and 16 Staphylococcus aureus isolates (80% of S. aureus isolates).
Discussion and Conclusion: In spite of appropriate lytic effect and also, high stability to environmental conditions, Due to its narrow host range to K. pneumoniae clinical isolates, it needs to exactly determination of its host range and efficacy of the phage against problematic strains of bacteria for using it as phage therapy candidate. Detailed survey of molecular characteristics of the bacteriophage is necessary.

Abstract Objective: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder among reproductive-age women, with a prevalence of 6–13%. The precise molecular network structure and central driver genes in PCOS pathogenesis remain incompletely understood.

Method: A total of 200 differentially expressed genes linked to PCOS were retrieved from multiple transcriptomic studies. Protein-protein interaction (PPI) networks were constructed using the STRING database (confidence score >0.40) and further analyzed in Cytoscape version 3.10.2. Hub genes were identified by CytoHubba plugin based on degree, betweenness, and closeness centrality measures. Pathway enrichment analysis was conducted using KEGG, Gene Ontology, Reactome, and WikiPathways databases; only terms with FDR <0.05 were considered significant.

Results: The resulting network consisted of 200 nodes and 4,979 edges. Four core genes—PIK3CA, PIK3CD, AKT1, and PIK3CB—were present in all eight highly dysregulated pathways (PI3K-Akt, AMPK, longevity regulating, AGE-RAGE, FoxO, insulin resistance, HIF-1, and TNF signaling), with PI3K-Akt showing the strongest enrichment (29 genes, FDR=3.28×10⁻¹â°). Three additional biomarkers (INSR, INS, IL6) overlapped in six pathways.

Conclusions: PIK3CA, PIK3CD, AKT1, and PIK3CB emerge as central molecular hubs and promising therapeutic targets in PCOS. Extensive pathway convergence on these nodes highlights the systemic metabolic-inflammatory basis of the disorder.

Abstract Nutlets of 20 taxa of Phlomoides, representing most of the currently recognized sections were examined by scanning electron microscopy. The basic shape of nutlets in most taxa studied is broadly ovate, but ovate, triangular and oblong ones can also be found in few species. Regarding the sculpturing pattern of nutlet surface, four basic types can be distinguished: reticulate, scalariform, ruminate and rugose. The reticulate type is the most common among the studied species, but the variation in alignment, size and shape of its composing cells provide further evidence that are useful as diagnostic characteristics. However, the type of sculpturing is more useful for separating species within the sections, rather than correlating them to each other. For example, sect. Filipendula show four different types of sculpturing. Therefore, nutlet microsculpturing is not useful in separating large natural groups like sections in this genus. It seems also that contrary to other genera of Lamiaceae, nutlet characters are of low phylogenetic value in this genus.

Abstract Ecosystems conservation requires knowing and monitoring habitats and study on their species. This goal is achieved when there is a suitable understanding of the current state of the environment and the living conditions of the species. In the present study, the distribution and biometry of adult and hatchlings of hawksbill turtles were investigated on Hendorabi Island, from March 2017 to July 2018. After observing turtles, species identification, biometry of adults including CCL and CCW and egg biometry including diameter and weight were measured. During this study, 28 turtles were examined biometrically. Some areas in the north and south of Hendorabi were identified as high-density nesting areas, while eastern part of the island was low-density nesting area. The average of CCL and CCW of the turtles were measured 70.38±3.27 and 64.84±2.53cm, respectively. On average, 87.6±21.7 eggs were laid per nest, and the highest and lowest numbers of eggs were recorded 110 and44, respectively. The number of normal and abnormal eggs were 74.2±9.02 and 13.6±5.81, respectively. Normal and abnormal eggs have an average diameters of 38.66±2.02 and 24.87±4.43mm and weights of 32.69±4.27 and 11.21±6.26gr respectively. In conclusion the hawksbill turtles of Hendorabi are smaller than those of other parts of the world.

Abstract Technological advances in three-dimensional (3D) cell culture have reshaped both basic research and translational applications in biomedicine. Unlike conventional two-dimensional (2D) systems, which cannot reproduce tissue architecture, biochemical gradients, or complex cell–cell and cell–matrix interactions, 3D platforms better recapitulate the native microenvironment and physiological dynamics that drive differentiation, gene expression, and drug responses. Within this family of models, spheroids—relatively simple, homogeneous multicellular aggregates—serve as practical and cost-effective tools for drug screening and cancer biology, whereas organoids derived from differentiated stem cells capture higher-order organotypic structure and function, making them especially valuable for disease modeling and precision medicine. Emerging technologies such as 3D bioprinting and organ-on-a-chip systems extend these capabilities by enabling precise architectural control, programmable microenvironments, and dynamic physiological cues under in vitro conditions. This review provides a critical, integrated analysis of principal 3D culture classes, compares their strengths and limitations, and examines recent progress in 3D bioprinting and organ-on-a-chip technologies with particular emphasis on bioink engineering, functional vascularization, and strategies to promote tissue maturation. By synthesizing current evidence, we outline a practical framework to guide the translation from simple laboratory constructs toward robust, human-relevant tissue models for research and therapeutic development.