The Society cited two pillars of Puglisi’s scientific career as the basis for the honor. The first is a body of sophisticated nuclear magnetic resonance, or NMR, studies that have advanced fundamental concepts of how RNA is recognized by other molecules. The second is a series of groundbreaking single-molecule experiments using fluorescence resonance energy transfer, commonly known as FRET, that have illuminated the inner workings of the ribosome, the molecular machine that translates genetic information into proteins. Together, these contributions have shaped how an entire generation of researchers thinks about RNA structure, dynamics, and function.

The award’s namesake carries deep personal significance in this case. Ignacio “Nacho” Tinoco, after whom the prize is named, was Puglisi’s mentor, and his contributions to the spectroscopic, thermodynamic, structural, and single-molecule study of RNA are considered foundational to the modern understanding of the physical principles governing macromolecules. Tinoco challenged the biophysics community to continually push the boundaries of fundamental understanding, and the award created in his memory recognizes scientists who carry that spirit forward. That Puglisi, a direct scientific descendant of Tinoco, should now receive the honor bearing his mentor’s name gives the 2027 announcement a rare narrative symmetry.

Biophysical Society President Karen Fleming of Johns Hopkins University praised Puglisi’s originality and the depth of his influence. “Jody is one of the most creative and innovative scientists in the field of translation,” Fleming said. “His recognition aptly honors the legacy of Ignacio “Nacho” Tinoco, Jody’s mentor, who challenged our community to continually push our fundamental understanding of biophysics.” Fleming added that Puglisi’s contributions are unique, noting that his research over the past decades has provided physical and structural foundations for RNA’s dynamic role in biology and has revealed many of the most complex yet fundamental properties of RNA-mediated biology.

To appreciate why Puglisi’s NMR work earned such recognition, it helps to understand what the technique makes possible. NMR spectroscopy allows researchers to probe the structure and motion of molecules at atomic resolution in solution, closer to the conditions of a living cell than many other structural methods. RNA, however, is notoriously difficult to study this way. Unlike proteins, many RNA molecules are flexible, dynamic, and conformationally heterogeneous, adopting ensembles of shapes rather than a single rigid structure. Extracting meaningful structural and dynamic information from such systems demands both methodological ingenuity and deep physical insight, qualities that have characterized Puglisi’s approach throughout his career.

Through decades of NMR studies, Puglisi and his collaborators helped establish how RNA molecules are recognized by binding partners, including proteins and small molecules. RNA recognition underlies a vast range of biological processes, from the regulation of gene expression to the action of antibiotics that target bacterial ribosomes. By defining the structural principles that govern these interactions, his work provided a conceptual framework that other laboratories have built upon, connecting atomic-level detail to the broader mechanics of molecular biology. This kind of fundamental knowledge is also a starting point for practical applications, since many drugs work by binding to RNA or to the molecular machines that interact with it.

The second strand of Puglisi’s recognized work, single-molecule FRET, represents a different but complementary way of seeing biology. In a FRET experiment, two fluorescent tags are placed on a molecule or molecular complex at carefully chosen positions. When the tags come close together, energy transfers between them, producing a signal that reports on the distance between the labeled sites. By monitoring individual molecules one at a time, researchers can watch molecular machines move in real time, capturing transient states and rare events that are invisible to methods that average signals over enormous populations of molecules. For something as dynamic as the ribosome, this capability is transformative.

Using these single-molecule approaches, Puglisi’s laboratory has dissected some of the most intricate behaviors of the ribosome during translation, the process by which messenger RNA is decoded into amino acid chains. The Society’s announcement specifically highlighted his studies of translational frameshifting, bypassing, stalling, and pausing during elongation. Frameshifting occurs when the ribosome slips by one or more nucleotides, reading the genetic message in a different frame and producing a different protein, a mechanism that certain viruses exploit and that cells use for regulation. Bypassing, meanwhile, allows a ribosome to skip over stretches of mRNA, while stalling and pausing during elongation influence how quickly and accurately proteins are made. Observing these events directly, one ribosome at a time, revealed kinetic details and intermediate states that population-level experiments simply could not resolve.

What makes this body of work especially significant is how it reframed the ribosome from a static structure into a dynamic machine. High-resolution structures had provided stunning snapshots of the ribosome at work, but the transitions between those states, their timing, and their probabilities remained largely hidden. Single-molecule FRET brought those dynamics into view, showing how the ribosome samples different conformations, how transfer RNAs move through the machine, and how the messenger RNA itself can be handled in unexpected ways. In doing so, Puglisi’s research connected the physical chemistry of molecular motion to one of biology’s most essential processes, the synthesis of every protein in every living cell.

The Biophysical Society, founded in 1958, is a professional scientific society established to lead a global community working at the interface of the physical and life sciences, across all levels of complexity, and to foster the dissemination of that knowledge. With roughly 6,000 members located around the world, the Society promotes growth in the field through its Annual Meeting, its publications, and its outreach activities, and its members teach and conduct research in colleges, universities, laboratories, government agencies, and industry. The Tinoco Award, presented within that community, honors a legacy of rigorous, quantitative inquiry into the molecules of life. When Puglisi accepts the 2027 prize in Philadelphia, the moment will recognize not only an individual career of technical brilliance and conceptual clarity, but also a scientific lineage running from Tinoco’s pioneering studies of RNA physical chemistry to the single-molecule era in which the movements of individual ribosomes can be watched as they read the genetic code.

Subject of Research: Physical chemistry of RNA and ribosome function recognized by the Biophysical Society's 2027 Ignacio Tinoco Award

Article Title: Joseph D. (Jody) Puglisi to receive the 2027 Ignacio Tinoco Award in Physical Chemistry of Macromolecules

Article References: Joseph D. (Jody) Puglisi to receive the 2027 Ignacio Tinoco Award in Physical Chemistry of Macromolecules. (n.d.). Original publication

Image Credits: AI Generated

DOI: Not provided

Keywords: Joseph Puglisi, Biophysical Society, Ignacio Tinoco Award, NMR spectroscopy, single-molecule FRET, RNA recognition, ribosome, translation, translational frameshifting, physical chemistry, macromolecules, Stanford University

The research team, led by G. Osthoff of the University of the Free State with collaborators from North-West University and the University of Pretoria, set out with three objectives: to describe quantified blood metabolites in healthy giraffes and southern white rhinoceroses, to test for sex-associated differences within each species, and to compare the two species using matrix-matched samples. The choice of animals was deliberate. Giraffes are foregut-fermenting ruminant browsers, while white rhinoceroses are hindgut-fermenting grazers, meaning their blood chemistry could plausibly differ because of species physiology, diet, and the microbial fermentation that occurs at opposite ends of the digestive tract.

The rhinoceros dataset was substantial: 45 apparently healthy, free-ranging white rhinoceroses, 31 females and 14 males, were sampled during routine management procedures on a single semi-extensive wildlife property in the Northern Cape Province over five days in May 2019. Each animal was immobilised with etorphine, hyaluronidase and azaperone, and blood was drawn from the posterior auricular vein into EDTA tubes, centrifuged within two hours, and stored at minus 80 degrees Celsius. The giraffe data came from 24 valid sampling occasions involving 19 animals at Rooipoort and Sandveld Nature Reserves, with serum collected in 2017 and EDTA plasma from six animals in 2018.

The analytical pipeline was exacting. Samples were filtered through 10 kDa membranes to remove proteins, mixed with a phosphate buffer in deuterium oxide containing an internal standard, and run on a Bruker Avance III HD 500 MHz spectrometer. Metabolites were annotated against pure-compound spectral libraries supplemented with two-dimensional J-resolved and COSY spectra, and identities were graded according to the Metabolomics Standards Initiative, with Level 1 assignments confirmed against reference standards. Concentrations were reported in micromoles per litre, and blank cells where no resonance was discernible were treated as non-detections rather than zeros, a distinction the authors stress is critical for honest interpretation.

The statistics were equally careful. Concentrations were log2-transformed and tested with Welch’s t-test, which does not assume equal variances, followed by Benjamini-Hochberg false-discovery correction across each family of metabolites. Effect sizes were expressed as ratios of geometric means with Hedges’ g confidence intervals, and robustness was probed with equal-variance t-tests, Mann-Whitney U tests, Monte Carlo permutation tests with 50,000 label permutations, and leave-one-out analyses that removed each animal in turn. Notably, the team explicitly declined to use PLS-DA, OPLS-DA, VIP-based feature selection or pathway enrichment as inferential evidence, positioning the study as a model of statistical restraint in a field often criticised for overinterpretation.

The first headline result concerned sex. In the design-compatible giraffe comparison, restricted to 2017 Rooipoort serum from 10 females and six males, no metabolite differed significantly even before correction for multiple testing; the smallest p value, for 3-hydroxyisobutyric acid, was just 0.070. The authors caution that this null result should not be read as proof that the sexes are metabolically equivalent, given the small sample and the absence of a predefined equivalence margin, but it aligns with growing evidence that sex effects are not universal across wildlife metabolomes.

The rhinoceros sex comparison told a more nuanced story. Nine metabolites met the primary false-discovery criterion, with females showing higher N-acetylglucosamine, creatinine, alanine, cholic acid, glycocholic acid, acetoacetic acid, isoleucine and valine, and lower allantoin than males. Yet only N-acetylglucosamine survived every sensitivity analysis, remaining significant under Welch, permutation and rank-based testing and in all 45 leave-one-out runs. Alanine and creatinine showed moderate support, while the remaining six findings were threshold-sensitive, hovering near the significance boundary. The result is a textbook demonstration of why multiplicity control and transparent robustness categories matter: without them, six fragile findings might have been reported alongside one genuinely solid one.

The interspecies comparison produced the study’s most dramatic pattern. Comparing EDTA plasma from six verified 2018 giraffes with all 45 rhinoceroses, 36 of 50 directly comparable metabolites differed significantly, and 28 formed a conservative core supported by every statistical test and every leave-one-giraffe-out analysis. Giraffe plasma contained markedly more pyruvic acid, glycine, allantoin and trimethylamine, and far less methanol, N-acetylglucosamine, acetic acid, 3-hydroxyisovaleric acid and methylamine. Most intriguingly, a coherent pattern emerged among branched-chain amino-acid-related compounds: giraffes showed lower leucine, 2-oxoisocaproic acid, 3-methyl-2-oxovaleric acid, 3-hydroxyisobutyric acid and 3-hydroxyisovaleric acid, consistent with a genuine difference in circulating branched-chain amino-acid chemistry between the species. Lower acetate and propionate in giraffes is biologically compatible with the contrast between foregut and hindgut fermentation, though the study did not measure feed intake or fermentation products directly.

The authors are admirably candid about the limits of their data. Species was perfectly confounded with NMR acquisition batch, since giraffe spectra were acquired in 2018 and rhinoceros spectra in 2021 with no shared quality-control material, so not every difference can be attributed to biology. Collection year was completely confounded with serum versus plasma in the giraffe dataset, precluding any inferential temporal comparison. The six-animal giraffe plasma group, while shown by leave-one-out analysis not to be driven by a single outlier, is small, and the concentrations are offered as exploratory baseline data rather than formal veterinary reference intervals, which would require dedicated reference-interval methodology. Adjustment for sex did not materially change the conservative core, ruling out the different sex ratios as an explanation for the broad pattern.

Why does this matter beyond the curiosity factor? Blood metabolomics is increasingly used in conservation medicine to detect disease, nutritional stress and environmental exposure in endangered species, from iron storage disease in Sumatran rhinoceroses to mitochondrial dysfunction in black rhinoceroses. Reliable baseline data from healthy animals are the foundation on which such diagnostic applications must be built, and they are scarce for wildlife. This study provides carefully curated concentration data for two keystone African herbivores, along with a methodological template, predefined contrasts, multiplicity correction, permutation testing and leave-one-out stability checks, that future wildlife metabolomics studies would do well to follow. The authors call for prospective work sampling both species with the same specimen matrix, balanced sexes, contemporaneous acquisition and shared quality controls. Until then, the branched-chain amino-acid signature separating the world’s tallest ruminant from its second-largest land mammal stands as an intriguing candidate for targeted investigation into how gut architecture writes itself into the blood.

Subject of Research: Comparative 1H-NMR blood metabolite profiling of giraffes and white rhinoceroses

Article Title: Comparative 1H-NMR profiling of plasma and serum metabolites in giraffes and white rhinoceroses

Article References: Osthoff, G., Mason, S., Schmidt, L., Tordiffe, A., & Deacon, F. (2026). Comparative 1H-NMR profiling of plasma and serum metabolites in giraffes and white rhinoceroses. Metabolomics, 22(5), Article 159. https://doi.org/10.1007/s11306-026-02535-0

Image Credits: AI Generated

DOI: 10.1007/s11306-026-02535-0

Keywords: metabolomics, giraffe, white rhinoceros, NMR spectroscopy, blood plasma, serum, branched-chain amino acids, wildlife physiology, conservation medicine, foregut fermentation, hindgut fermentation, South Africa

Pyrazoles are five-membered rings containing two adjacent nitrogen atoms, and they have been a mainstay of medicinal chemistry since their discovery by Ludwig Knorr in 1883. Their ring positions can be readily functionalized, allowing chemists to tune properties such as lipophilicity, metabolic stability, and target binding. Pyrazole derivatives have been investigated as anti-inflammatory, antifungal, antidiabetic, and anticancer agents, and they also serve as ligands in catalysis and materials science. Yet despite this rich history, relatively few studies have combined the pyrazole framework with organophosphorus groups, a class of substituents known for their unique electronic and steric properties and their proven value in pharmaceuticals and agrochemicals.

The research team, Dhiab Jabli and Mohamed Lotfi Efrit of the Laboratory of Selective Organic and Heterocyclic Synthesis at El Manar University in Tunis, addressed this gap by exploiting phosphonohydrazines as versatile building blocks. These reagents, which pair a nucleophilic hydrazine unit with a phosphoryl or thiophosphoryl group, react efficiently with activated alpha,beta-unsaturated nitriles under mild conditions. In the reported synthesis, two phosphonohydrazine precursors, a diethyl phosphonate and an O,O-diphenyl phosphonothioate, were reacted with a series of five activated nitrile substrates in ethanol with a catalytic amount of acetic acid under reflux for nine hours. The result was a library of ten N-phosphorylated 5-aminopyrazoles isolated in good to excellent yields ranging from 64 to 83 percent.

The proposed mechanism follows a well-established logic for electron-deficient olefins. The terminal amino group of the phosphonohydrazine first performs a Michael-type nucleophilic attack on the beta-carbon of the unsaturated nitrile. The remaining nitrogen then cyclizes intramolecularly onto the cyano group, closing the pyrazole ring, and a final tautomerization delivers the thermodynamically favored 5-amino-4-cyanopyrazole product. The authors are careful to note that this pathway is a plausible proposal grounded in literature precedents rather than definitive mechanistic proof, since no intermediates were isolated or directly observed. The absence of splitting in the nuclear magnetic resonance spectra, however, indicates that each reaction produces a single isomer, validating the selectivity of the route.

Structural confirmation relied on a comprehensive battery of analytical techniques. Fourier-transform infrared spectroscopy revealed the characteristic nitrile stretch near 2240 wavenumbers, amino absorptions between 3435 and 3355 wavenumbers, and phosphoryl bands consistent with P=O or P=S functionality. Proton and carbon-13 NMR spectra in DMSO-d6 mapped every substituent, with carbon-phosphorus coupling constants of roughly 23 to 25 hertz providing direct evidence of bonding between the ring framework and the phosphoryl substituent. Most diagnostic were the phosphorus-31 NMR chemical shifts: the phosphonate derivatives resonated between -1.34 and -2.63 parts per million, while the thiophosphonate analogues appeared far downfield between 49.78 and 61.63 parts per million, a deshielding attributed to the anisotropic environment of the pyrazole ring and to intramolecular hydrogen bonding between the phosphoryl group and the primary amine. Liquid chromatography tandem mass spectrometry and elemental analysis completed the structural picture, with characteristic fragment ions at mass-to-charge ratios of 137 and 248 corresponding to the phosphonate and thiophosphonate cores respectively.

To probe biological activity, the team selected four representative compounds, 3a, 3e, 4a, and 4e, chosen to capture the principal structural variations in the series: phosphonate versus phosphonothioate functionality and different substituents at the C-3 position of the ring. Using standard disc diffusion assays on Mueller-Hinton agar with bacterial suspensions standardized to 0.5 McFarland, the compounds were tested at 100, 150, and 200 micrograms per milliliter against four clinically relevant strains: Salmonella typhimurium, Enterococcus faecium, Staphylococcus aureus, and Escherichia coli. Ampicillin served as the reference antibiotic and DMSO-treated discs as negative controls, with all experiments performed in triplicate and analyzed by one-way ANOVA followed by Tukey’s post hoc test.

The results were striking. Activity was consistently stronger against Gram-negative bacteria than Gram-positive ones, with a sensitivity order of S. typhimurium, S. aureus, E. faecium, and finally E. coli, which showed the largest inhibition zones, reaching up to 22 millimeters at the highest concentration. This Gram-negative potency is noteworthy because the outer membrane of these bacteria typically blocks many conventional antibiotics. The fact that the N-phosphorylated aminopyrazoles retained measurable activity suggests they may bypass or disrupt this defensive barrier. While the compounds did not surpass ampicillin in overall potency, their activity profile points to a mechanism of action potentially complementary to beta-lactam drugs.

Structure-activity analysis revealed clear trends. Phosphonothioate derivatives generally outperformed their phosphonate analogues, a difference the authors attribute to the higher polarizability of sulfur, which may strengthen hydrophobic interactions with bacterial membranes and facilitate cellular penetration. The single most active compound was 4e, which combines the thiophosphoryl group with a methylthio substituent at C-3, a pairing that increases lipophilicity and reshapes the electronic distribution of the scaffold. Its minimum bactericidal concentration to minimum inhibitory concentration ratio fell below 2, indicating genuinely bactericidal rather than merely bacteriostatic action, and its MIC and MBC values approached those of ampicillin. By contrast, compound 3e showed reduced activity, likely due to steric hindrance and diminished aqueous solubility.

The molecular explanation for this potency is multifactorial. The nitrogen-rich pyrazole ring serves as a hydrogen-bond donor and acceptor network capable of engaging bacterial enzymes, nucleic acids, and membrane phospholipids, potentially disrupting DNA replication, cell-wall biosynthesis, and metabolic function. The amino and imine functionalities enhance electrostatic complementarity with polar enzyme cavities, while the phosphoryl group can coordinate essential metal cofactors such as zinc and magnesium ions, inhibiting metalloenzymes involved in oxidative stress regulation. Additionally, the conjugated pi-system of the heteroaromatic scaffold may stack with nucleobases in microbial DNA, interfering with replication and transcription. In compound 4e, these electronic effects, hydrogen-bonding capacity, and balanced lipophilicity appear to act synergistically, maximizing membrane permeability, enzyme affinity, and intracellular stability.

The authors emphasize that this biological evaluation is a preliminary screening, but it establishes an initial structure-activity relationship that can guide rational design. Future work will extend testing to the remaining derivatives, employ molecular docking and computational studies against bacterial targets to clarify the mechanism of action, and evaluate activity against multidrug-resistant strains. For a field urgently seeking alternatives to failing antibiotics, phosphorus-functionalized pyrazoles have now earned a place on the shortlist of promising scaffolds, and the Tunisian team’s efficient synthetic route provides the practical chemistry needed to explore that potential at scale.

Subject of Research: Synthesis of N-phosphorylated 5-aminopyrazole derivatives and evaluation of their antimicrobial activity

Article Title: Phosphonohydrazine mediated synthesis of N phosphorylated 5 aminopyrazoles with structural characterization and antimicrobial activity

Article References: Phosphonohydrazine mediated synthesis of N phosphorylated 5 aminopyrazoles with structural characterization and antimicrobial activity. (n.d.). https://doi.org/10.1007/s44371-026-00949-2

Image Credits: AI Generated

DOI: 10.1007/s44371-026-00949-2

Keywords: pyrazole, organophosphorus, antimicrobial activity, Escherichia coli, phosphonohydrazine, heterocyclic synthesis, antibiotic resistance, NMR spectroscopy, structure-activity relationship, medicinal chemistry, Gram-negative bacteria, thiophosphonate