Comprehensive deciphering the alternative splicing patterns involved in leaf morphogenesis of Liriodendron chinense.

Alternative splicing (AS), a pivotal post-transcriptional regulatory mechanism, profoundly amplifies diversity and complexity of transcriptome and proteome. Liriodendron chinense (Hemsl.) Sarg., an excellent ornamental tree species renowned for its distinctive leaf shape, which resembles the mandarin jacket. Despite the documented potential genes related to leaf development of L. chinense, the underlying post-transcriptional regulatory mechanisms remain veiled. Here, we conducted a comprehensive analysis of the transcriptome to clarify the genome-wide landscape of the AS pattern and the spectrum of spliced isoforms during leaf developmental stages in L. chinense. Our investigation unveiled 50,259 AS events, involving 10,685 genes (32.9%), with intron retention as the most prevalent events. Notably, the initial stage of leaf development witnessed the detection of 804 differentially AS events affiliated with 548 genes. Although both differentially alternative splicing genes (DASGs) and differentially expressed genes (DEGs) were enriched into morphogenetic related pathways during the transition from fishhook (P2) to lobed (P7) leaves, there was only a modest degree of overlap between DASGs and DEGs. Furthermore, we conducted a comprehensively AS analysis on homologous genes involved in leaf morphogenesis, and most of which are subject to post-transcriptional regulation of AS. Among them, the AINTEGUMENTA-LIKE transcript factor LcAIL5 was characterization in detailed, which experiences skipping exon (SE), and two transcripts displayed disparate expression patterns across multiple stages. Overall, these findings yield a comprehensive understanding of leaf development regulation via AS, offering a novel perspective for further deciphering the mechanism of plant leaf morphogenesis.

The hydrophobic cluster on the surface of protein is the key structural basis for the SDS-resistance of chondroitinase VhChlABC.

The application of chondroitinase requires consideration of the complex microenvironment of the target. Our previous research reported a marine-derived sodium dodecyl sulfate (SDS)-resistant chondroitinase VhChlABC. This study further investigated the mechanism of VhChlABC resistance to SDS. Focusing on the hydrophobic cluster on its strong hydrophilic surface, it was found that the reduction of hydrophobicity of surface residues Ala181, Met182, Met183, Ala184, Val185, and Ile305 significantly reduced the SDS resistance and stability. Molecular dynamics (MD) simulation and molecular docking analysis showed that I305G had more conformational flexibility around residue 305 than wild type (WT), which was more conducive to SDS insertion and binding. The affinity of A181G, M182A, M183A, V185A and I305G to SDS was significantly higher than that of WT. In conclusion, the surface hydrophobic microenvironment composed of six residues was the structural basis for SDS resistance. This feature could prevent the binding of SDS and the destruction of hydrophobic packaging by increasing the rigid conformation of protein and reducing the binding force of SDS-protein. The study provides a new idea for the rational design of SDS-resistant proteins and may further promote chondroitinase research in the targeted therapy of lung diseases under the pressure of pulmonary surfactant. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-023-00201-1.

Metabolic profiles of Fufang Xiling Jiedu capsule in rats by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry.

Fufang Xiling Jiedu capsule (FXJC), a traditional Chinese medicine that evolved from "Yinqiao Powder", is widely used for the treatment of cold and influenza. However, due to a lack of in vivo metabolism research, the chemical components responsible for the therapeutic effects still remain unclear. Hence, this study aimed to describe the metabolic profiles of the FXJC in rat plasma, urine, and feces. A combined data mining strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry was employed and 201 xenobiotics, including 117 prototype components and 84 metabolites were detected. Phenolic acids, flavonoids, triterpenes, and lignans were prominent ingredients absorbed in vivo, and the major metabolic pathways of the detected metabolites were glucuronidation, sulfation, methylation, and oxidation. This is the first systematic study on the metabolism of the FXJC in vivo, providing valuable information for future studies on the efficacy, toxicity, and mechanism of the FXJC.

Adenine nucleotide translocase 2 (Ant2) is required for individualization of spermatogenesis of Drosophila melanogaster.

Successful completion of spermatogenesis is crucial for the perpetuation of the species. In Drosophila, spermatid individualization, a process involving changes in mitochondrial structure and function is critical to produce functional mature sperm. Ant2, encoding a mitochondrial adenine nucleotide translocase, is highly expressed in male testes and plays a role in energy metabolism in the mitochondria. However, its molecular function remains unclear. Here, we identified an important role of Ant2 in spermatid individualization. In Ant2 knockdown testes, spermatid individualization complexes composed of F-actin cones exhibited a diffuse distribution, and mature sperms were absent in the seminal vesicle, thus leading to male sterility. The most striking effects in Ant2-knockdown spermatids were decrease in tubulin polyglycylation and disruption of proper mitochondria derivatives function. Excessive apoptotic cells were also observed in Ant2-knockdown testes. To further investigate the phenotype of Ant2 knockdown in testes at the molecular level, complementary transcriptome and proteome analyses were performed. At the mRNA level, 868 differentially expressed genes were identified, of which 229 genes were upregulated and 639 were downregulated induced via Ant2 knockdown. iTRAQ-labeling proteome analysis revealed 350 differentially expressed proteins, of which 117 proteins were upregulated and 233 were downregulated. The expression of glutathione transferase (GstD5, GstE5, GstE8, and GstD3), proteins involved in reproduction were significantly regulated at both the mRNA and protein levels. These results indicate that Ant2 is crucial for spermatid maturation by affecting mitochondrial morphogenesis.

RpS3 Is Required for Spermatogenesis of Drosophila melanogaster.

Ribosomal proteins (RPs) constitute the ribosome, thus participating in the protein biosynthesis process. Emerging studies have suggested that many RPs exhibit different expression levels across various tissues and function in a context-dependent manner for animal development. Drosophila melanogaster RpS3 encodes the ribosomal protein S3, one component of the 40S subunit of ribosomes. We found that RpS3 is highly expressed in the reproductive organs of adult flies and its depletion in male germline cells led to severe defects in sperm production and male fertility. Immunofluorescence staining showed that RpS3 knockdown had little effect on early germ cell differentiation, but strongly disrupted the spermatid elongation and individualization processes. Furthermore, we observed abnormal morphology and activity of mitochondrial derivatives in the elongating spermatids of RpS3-knockdown testes, which could cause the failure of axoneme elongation. We also found that RpS3 RNAi inhibited the formation of the individualization complex that takes charge of disassociating the spermatid bundle. In addition, excessive apoptotic cells were detected in the RpS3-knockdown testes, possibly to clean the defective spermatids. Together, our data demonstrated that RpS3 plays an important role in regulating spermatid elongation and individualization processes and, therefore, is required for normal Drosophila spermatogenesis.

LcCCL28-25, Derived from Piscine Chemokine, Exhibits Antimicrobial Activity against Gram-Negative and Gram-Positive Bacteria In Vitro and In Vivo.

Antimicrobial peptides (AMPs) are currently recognized as potentially promising antibiotic substitutes. Fish are an important seawater/freshwater medicinal biological resource, and the antimicrobial peptides and proteins that are key components of their innate immune systems are potential candidates for the development of novel antibacterial agents. The rainbow trout Oncorhynchus mykiss chemokine CK11 (omCK11), classified in the C-C motif chemokine ligand 27/28 (CCL27/28) family, is the only CC-type chemokine reported to play a direct antibacterial role in the immune response; however, its antibacterial domain remains unknown. In this study, we analyzed the structure-activity relationship of omCK11 and identified the antibacterial C-terminal domain. Additionally, we performed structure-function analyses of CCL27/28 proteins from different, representative freshwater and seawater fishes, revealing their shared C-terminal antibacterial domains. Surprisingly, a synthesized cationic peptide (named lcCCL28-25), derived from the large yellow croaker Larimichthys crocea CCL28, exhibited broad-spectrum and the most acceptable bactericidal activity, as well as antibiofilm activity and negligible hemolytic and cytotoxic activity in vitro. Additionally, lcCCL28-25 conferred a protective effect in the thighs of neutropenic mice infected with Staphylococcus aureus. SYTOX green fluorescence and electron microscopy experiments revealed that lcCCL28-25 was capable of rapidly destroying the integrity and permeability of the bacterial cell membrane. Overall, this study aided in the advancement of antibacterial CC-type chemokine research and also suggested a new strategy for exploring novel AMPs. Additionally, the efficacy of lcCCL28-25 in in vivo antibacterial activity in a mammalian model revealed that this compound could be a promising agent for the development of peptide-based antibacterial therapeutics. IMPORTANCE The primary function of chemokines has been described as recruiting and activating leukocytes to participate in the immune response. Some chemokines are also broad-spectrum antibacterial proteins in mammals. The Oncorhynchus mykiss chemokine CK11 (omCK11) is the first reported and currently the only CC-type antibacterial chemokine. The present study identified the antibacterial domain of omCK11. Structure-function analysis of various fish CCL27/28 proteins identified a novel antibacterial peptide (lcCCL28-25) from Larimichthys crocea CCL28 that exhibited broad-spectrum and the most acceptable bactericidal activity in vitro, as well as a protective effect in a Staphylococcus aureus infection mouse model. The antibacterial mechanisms included membrane disruption and permeation. This study advanced the field of antibacterial chemokine research in fish and also suggested a new strategy for exploring novel AMPs. The novel peptide lcCCL28-25 may prove to be an effective antibacterial agent.

The influence of dexmedetomidine on the emergence agitation of pediatric patients after the operations of sense organs under general anesthesia using sevoflurane.

BACKGROUND: We investigated the influence of dexmedetomidine on the emergence agitation of pediatric patients after ophthalmologic operation under general anesthesia using sevoflurane. METHODS: We selected 90 patients that were administered pediatric ophthalmologic operation for the study. The patients were randomly divided into 3 groups according to the administration way of drugs, i.e. the normal saline group (group S, N.=30), the midazolam group (group M, N.=30) and the dexmedetomidine group (group D, N.=30). For all patients, anesthesia induction was performed using sevoflurane before anesthesia, and the anesthesia was maintained in the operation with a combination of sevoflurane and remifentanil; laryngeal mask airway (LMA) was used for assisted ventilation. Ten minutes before the end of operation, 15 mL of 0.9% normal saline, 0.05 mg/kg of midazolam and 0.5 µg/kg of dexmedetomidine were administered to group S, group M and group D, respectively. After the operation, we observed the awakening time, time of the LMA removal as well as the recovery time in the Post Anesthesia Care Unit (PACU) of patients in all three groups. We evaluated the postoperative condition of sedation and agitation of the patients using Ramsay Sedation Scale, 5-point scale and Pediatric Anesthesia Emergence Deliriums Scale (PAED) and performed statistical analysis. RESULTS: In the comparisons of awakening time, time of the LMA removal as well as the recovery time, we found that group M was the longest sequentially followed by group D and group S with statistically significant differences (P<0.05). While the comparison of the scores of Ramsay Sedation Scale revealed that group D scored highest followed by group M and group S with statistically significant differences (P<0.05), both of the comparisons of the scores of 5-point scale and PAED Scale showed that group D scored the lowest, followed by group M and group S in sequence with statistically significant differences (P<0.05). CONCLUSIONS: Dexmedetomidine can significantly lower the incidence of emergence agitation of pediatric patients after the ophthalmologic operation under sevoflurane anesthesia.

The Ubiquitin Conjugating Enzyme UbcD1 is Required for Notch Signaling Activation During Drosophila Wing Development.

Notch signaling pathway plays crucial roles in animal development. Protein ubiquitination contributes to Notch signaling regulation by governing the stability and activity of major signaling components. Studies in Drosophila have identified multiple ubiquitin ligases and deubiquitinating enzymes that modify Notch ligand and receptor proteins. The fate of ubiquitinated substrates depend on topologies of the attached ubiquitin chains, which are determined by the ubiquitin conjugating enzymes (E2 enzymes). However, which E2 enzymes participate in Notch signal transduction remain elusive. Here, we report that the E2 enzyme UbcD1 is required for Notch signaling activation during Drosophila wing development. Mutations of UbcD1 lead to marginal nicks in the adult wing and reduction of Notch signaling targets expression in the wing imaginal disc. Genetic analysis reveal that UbcD1 functions in the signaling receiving cells prior to cleavage of the Notch protein. We provide further evidence suggesting that UbcD1 is likely involved in endocytic trafficking of Notch protein. Our results demonstrate that UbcD1 positively regulates Notch signaling and thus reveal a novel role of UbcD1 in development.

Biochemical Characterization of a Novel Exo-Type PL7 Alginate Lyase VsAly7D from Marine Vibrio sp. QY108.

Brown algae is a kind of renewable resource for biofuels production. As the major component of carbohydrate in the cell walls of brown algae, alginate can be degraded into unsaturated monosaccharide by exo-type alginate lyases, then converted into 4-deoxy-L-erythro-5-hexoseulose uronate (DEH) by a non-enzyme reaction, which is an important raw material for the preparation of bioethanol. In our research, a novel exo-type alginate lyase, VsAly7D, belonging to the PL7 family was isolated from marine bacterium Vibrio sp. QY108 and recombinantly expressed in Escherichia coli. The purified VsAly7D demonstrated the highest activity at 35 °C, whereas it still maintained 46.5% and 83.1% of its initial activity at 20 °C and 30 °C, respectively. In addition, VsAly7D exhibited the maximum activity under alkaline conditions (pH 8.0), with the simultaneously remaining stability between pH 8.0 and 10.0. Compared with other reported exo-type enzymes, VsAly7D could efficiently degrade alginate, poly-ß-D-mannuronate (polyM) and poly-α-L-guluronate (polyG) with highest specific activities (663.0 U/mg, 913.6 U/mg and 894.4 U/mg, respectively). These results showed that recombinant VsAly7D is a suitable tool enzyme for unsaturated alginate monosaccharide preparation and holds great promise for producing bioethanol from brown algae.

Selection of Muscle-Activity-Based Cost Function in Human-in-the-Loop Optimization of Multi-Gait Ankle Exoskeleton Assistance.

Using "human-in-the-loop" (HIL) optimization can obtain suitable exoskeleton assistance patterns to improve walking economy. However, there are differences in these patterns under different gait conditions, and currently most HIL optimizations use metabolic cost, which requires long periods to be estimated for each control law, as the physiological objective to minimize. We aimed to construct a muscle-activity-based cost function and to find the appropriate initial assistance patterns in HIL optimization of multi-gait ankle exoskeleton assistance. One healthy subject walked assisted by an ankle exoskeleton under nine gait conditions and each condition was the combination of different walking speeds, ground slopes and load weights. Ten assistance patterns were provided for the subject under each gait condition. Then we constructed a cost function based on surface electromyography signals of four lower leg muscles and select the muscle weight combination by using particle swarm optimization algorithm to compose the cost function with maximum differences between different assistance patterns. The mean weights of medial gastrocnemius, lateral gastrocnemius, soleus and tibialis anterior activity under all gait conditions are 0.153, 0.104, 0.953 and 0.145, respectively. Then we verified the effectiveness of this cost function by optimization and validation experiments conducted on four subjects. Our results are expected to guide the selection of muscle-activity-based cost functions and improve the time efficiency of HIL optimization.

Investigation of the role of miR-221 in diabetic peripheral neuropathy and related molecular mechanisms.

BACKGROUND: Diabetic peripheral neuropathy (DPN) is one of the most common complications of diabetes, but the molecular mechanisms of DPN are still unclear. OBJECTIVES: To investigate the role of miR-221 in DPN and the related molecular mechanisms. MATERIAL AND METHODS: Streptozotocin (STZ) was used to establish an in vivo DPN model. An in vitro DPN model was established using high glucose-induced SH-SY5Y cells. The pain condition of rats was measured by evaluating the 50% paw withdrawal threshold (PWT) and paw withdrawal latency (PWL). Serum exosomes were extracted and identified. Expression of miR-221 in serum exosomes and serum SOCS3 expression were determined using reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Western blotting was used to measure the protein levels of SOCS3, bradykinin (BK) and prostaglandin E2 (PEG2). The dual luciferase reporter assay was performed to confirm SOCS3 3'-UTR as a target of miR-221. The serum or cell supernatant levels of PEG2, BK, interleukin (IL)-6, IL-1ß, and tumor necrosis factor alpha (TNF-α) were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Induction of the lenti-miR-221 inhibitor significantly decreased the expression of miR-221 in DPN rats. Both 50% PWT and PWL values were markedly decreased in DPN rats. When miR-221 was inhibited, the 50% PWT and PWL values were both significantly increased. Knockdown of miR-221 significantly increased the expression of SOCS3 and decreased the expression of NF-κB. Furthermore, knockdown of miR-221 remarkably decreased the expression of PEG2, BK, IL-6, IL-1ß, and TNF-α in both STZ-treated DPN rats and high glucose-induced SH-SY5Y cells, which was reversed by inhibition of SOCS3. The dual luciferase reporter assay showed that miR-221 directly targeted and negatively regulated SOCS3. CONCLUSIONS: Inhibition of miR-221 can reduce pain and decrease expression of inflammatory factors through targeting SOCS3 in DPN.

Characterization of a new mastermind allele identified from somatic mosaic screen.

The Notch signaling pathway is highly conserved and regulates various fundamental development events. Activation of Notch signaling relies on production of the Notch intracellular domain (NICD), which assembles a transcription factor complex to turn on down-stream targets expression. The mastermind (mam) gene encodes an essential co-activator that permits NICD activity in the cell nucleus. During a somatic mosaic screen in Drosophila, an uncharacterized gene l(2)S9998 is identified as a positive regulator of the Notch signaling pathway. Genetic analysis demonstrates that l(2)S9998 functions at the level of transcriptional activation of Notch targets in the signal receiving cells. Whole genome sequencing reveals that l(2)S9998 is a novel allele of the mam gene, which is further confirmed by complementation tests. Along with three molecularly defined transposon insertions isolated from the screen, four mutants of mam are shown to modulate Notch signaling during fly wing development. Our analysis provides additional genetic resources for understanding mam function and Notch signaling regulation.

Decapentaplegic signaling regulates Wingless ligand production and target activation during Drosophila wing development.

The Decapentaplegic (Dpp) and Wingless (Wg) signaling pathways are essential for animal development. However, how these two signals are integrated in distinct tissues is not fully understood. Here, we describe a novel mode of Dpp-Wg crosstalk during Drosophila wing development. We show that the canonical Dpp signaling is required for Wg target gene activation. In addition, Dpp signaling inhibits the transcription of wg through the schnurri (shn) repressor complex. A Dpp-responsive shn/pMad/Med silencer element (SSE) is identified in the genomic loci of the wg gene. ChIP analysis suggests that shn interacts with this element in vivo. Our findings support a model in which Dpp signaling plays a dual role in transcriptional regulation of both the wg gene and downstream targets.

The conserved mitochondrial genomes of Drosophila mercatorum (Diptera: Drosophilidae) with different reproductive modes and phylogenetic implications.

Fruit flies (Drosophilidae: Drosophila) are commonly found in daily life and have long been used as model organisms in biology researches. Drosophila mercatorum is one important member of the Drosophila genus and has been used to study centrosome assembly of cells. In this study, we sequenced and analyzed the mitochondrial genome (mitogenome) of D. mercatorum, finding that it contains the typical structure of 37 genes and a control region. The arrangement of mitochondrial genes is in accordance with that in other Drosophila species, which is considered the ancestral organization of insects' mitogenomes. Phylogenetic analyses were performed based on 23 species of Drosophila. Our results supported two monophyletic subgenera, Drosophila and Sophophora, except for D. willistoni which was presented as an early offshoot of Drosophila. The topology ((D. yakuba + D. erecta) + D. melanogaster) was supported. We further compared the mitogenomes of parthenogenesis and sexual reproduction strains of D. mercatorum. However, only one synonymous mutation in COI gene was identified, indicating mitogenomic evolution is not strongly correlated with the different reproductive modes of this species. Taken together, our results demonstrate that mitogenome is an effective molecular marker that can be further used in phylogenetic studies of Drosophila and other organisms.

Comparison of desflurane and sevoflurane on postoperative recovery quality after tonsillectomy and adenoidectomy in children.

Comparison of desflurane and sevoflurane on the postoperative recovery quality after tonsillectomy and adenoidectomy in children was carried out. A retrospective analysis was performed on the medical records of 165 children who underwent tonsil and adenoid radiofrequency ablation under low-temperature plasma and were admitted to the Xuzhou Children's Hospital, Xuzhou Medical University from February 2014 to May 2017. In total, 79 children with sevoflurane anesthesia were in the sevoflurane group, and 86 children with desflurane anesthesia in the desflurane group. The non-invasive blood pressure (NIBP), heart rate (HR) and oxygen saturation (SpO2) level, the postoperative sedation (Ramsay) scores, the modified objective pain score (MOPS) of children were recorded. The pediatric anesthesia emergence delirium (PAED) scores of children were recorded. Children in the sevoflurane group had longer operation time, anesthesia time, extubation time and coincidence time than those in the desflurane group (P<0.05). At the beginning of operation (t1), 10 min after operation (t2), at the time of entering anesthesia recovery room (t3), at the time of tracheal catheter extubated (t4), 10 min after extubation (t5), and at the time of leaving the anesthesia recovery room (t6), children in the sevoflurane had higher NISBP and NIDBP, lower HR than those in the desflurane group (P<0.05). At the time of the tracheal catheter extubation (c2), 10 min after extubation (c3), 30 min after extubation (c4), children in the sevoflurane group had lower Ramsay scores and higher PAED scores than those in the desflurane group (P<0.05). More suitable as an anesthetic maintenance drug for tonsillectomy and adenoidectomy in children, desflurane has a better anesthetic effect and is safer. In addition, children with desflurane anesthesia have high postoperative recovery quality and quick recovery in the short term, with better sedative and analgesic effects. Therefore, it is worthy of promotion in clinic practice.

Efficacy and safety of propofol in preventing emergence agitation after sevoflurane anesthesia for children.

Application of propofol in preventing emergence agitation after sevoflurane anesthesia in children was evaluated. Clinical data of 200 children who received sevoflurane anesthesia in Children's Hospital of Xuzhou Medical University were retrospectively analyzed. Among them, 120 patients who received inhaled sevoflurane for pediatric anesthesia and intravenous infusion of propofol (2 mg/kg) were included in observation group. The remaining 80 cases who were directly anesthetized by sevoflurane alone were the control group. T PAED scores, modified Aldrete scores, extubation time, PACU time and adverse reactions (gastrointestinal tract and respiratory response) were analyzed and compared between the control and observation group. PAED scores, extubation time, PACU time and incidence of adverse reactions were significantly lower in observation than in control group, and the modified Aldrete scores were higher in observation than in control group (P<0.05). Spearman's correlation analysis showed that the PAED scores were negatively correlated with modified Aldrete scores and positively correlated with extubation time. There was positive correlation between the PACU time and incidence of adverse reactions and between the PAED scores and extubation time. There was negative correlation between PACU time and incidence of adverse reactions and between Aldrete scores and extubation time (P<0.05). Therefore, we conclude that propofol can be used to prevent agitation after sevoflurane anesthesia in children.

Antiretroviral therapy in HIV-infected children.

The pediatric highly-active antiretroviral therapy (HAART) has shown enormous progress in recent years but still, a high percentage of children continue to experience treatment failure. The main causes found responsible are the development of drug resistance, inadequate dosing and poor adherence. This review is aimed at exploring the current status of antiretroviral therapy in children with focus on the interaction between disease, drug and patient behavior, all of which are strongly correlated .An overview of HIV viral characteristics and of the available antiretroviral drug classes currently combined to avoid development of resistance has been also discussed.

Broadband quantitative NQR for authentication of vitamins and dietary supplements.

We describe hardware, pulse sequences, and algorithms for nuclear quadrupole resonance (NQR) spectroscopy of medicines and dietary supplements. Medicine and food safety is a pressing problem that has drawn more and more attention. NQR is an ideal technique for authenticating these substances because it is a non-invasive method for chemical identification. We have recently developed a broadband NQR front-end that can excite and detect 14N NQR signals over a wide frequency range; its operating frequency can be rapidly set by software, while sensitivity is comparable to conventional narrowband front-ends over the entire range. This front-end improves the accuracy of authentication by enabling multiple-frequency experiments. We have also developed calibration and signal processing techniques to convert measured NQR signal amplitudes into nuclear spin densities, thus enabling its use as a quantitative technique. Experimental results from several samples are used to illustrate the proposed methods.

Authentication of Medicines Using Nuclear Quadrupole Resonance Spectroscopy.

The production and sale of counterfeit and substandard pharmaceutical products, such as essential medicines, is an important global public health problem. We describe a chemometric passport-based approach to improve the security of the pharmaceutical supply chain. Our method is based on applying nuclear quadrupole resonance (NQR) spectroscopy to authenticate the contents of medicine packets. NQR is a non-invasive, non-destructive, and quantitative radio frequency (RF) spectroscopic technique. It is sensitive to subtle features of the solid-state chemical environment and thus generates unique chemical fingerprints that are intrinsically difficult to replicate. We describe several advanced NQR techniques, including two-dimensional measurements, polarization enhancement, and spin density imaging, that further improve the security of our authentication approach. We also present experimental results that confirm the specificity and sensitivity of NQR and its ability to detect counterfeit medicines.