Structural and dynamic characterization of the heterodimeric and homodimeric complexes of distamycin and 1-methylimidazole-2-carboxamide-netropsin bound to the minor groove of DNA

NMR spectroscopy combined with molecular modeling was used to characterize a heterodimeric complex with Dst and 2-ImN bound in the minor groove of d(GCCTAACAAGG)•d(CCTTGTTAGGC) (1:1:1 2-ImN•Dst•DNA complex). The imidazole-pyrrole-pyrrole ligand 2-ImN spans 5'-GTTA-3' of the TAACA•TGTTA binding site with the imidazole nitrogen specifically recognizing the guanine amino group. The Dst ligand lies along the 5'-AACA-3' sequence and complements the 2-ImN ligand in the formation of the antiparallel side-by-side heterodimeric complex. Titrations of the same site with Dst or 2-ImN alone yield homodimeric complexes (2:1 ligand.DNA) of lower stability than the 1:1:1 2-ImN•Dst•DNA complex. Dst and 2-ImN binding to d(CGCAAACTGGC)•d(GCCAGTTTGCG) was also investigated. The 1:1:1 2-ImN•Dst•DNA complex is again the most stable complex with the AAACT•AGTTT site and is similar to the TAACA•TGTTA complex. No monomeric binding of either 2-ImN or Dst was observed to either site

NMR characterization of hairpin polyamide complexes with the minor groove of DNA

Polyamides containing N-methylimidazole (Im) and N-methylpyrrole (Py) amino acids can be combined in antiparallel side-by-side dimeric complexes for sequence-specific recognition in the minor groove of DNA. Covalently linking polyamide subunits has led to designed ligands with both increased affinity and specificity. Simple aliphatic amino acid linkers serve as internal guide residues for turn vs extended binding in a head-to-tail-linked polyamide motif. Polyamides of sequence composition ImPyPy-X-PyPyPy containing linkers of incremental length (X = 3-aminopropionic acid (β), 4-aminobutyric acid (γ), or 5-aminovaleric acid (δ)) in complex with an undecamer DNA duplex containing a 5'-(A,T)G(A,T)(3)-3' target site were structurally characterized using NMR spectroscopy. Previous quantitative DNase I footprinting studies identified gamma as the highest affinity of these "turn" linkers. NMR titrations and 2D NOESY data combined with restrained molecular modeling reveal that polyamides with β, γ, and δ linkers all may adopt a hairpin structure. Modeling supports the idea that the linkers in the βand δcomplexes adopt an energetically less favorable turn geometry than the γlinker and confirms that the three-carbon γ linker is sufficient and optimal for the hairpin conformation

Youth Mental Health First Aid Training: Impact on the Ability to Recognize and Support Youth Needs.

Youth Mental Health First Aid (YMHFA) trains individuals who regularly interact with youth to identify youth experiencing mental health challenges. Several studies demonstrate positive training impacts, but few assess whether the training equally impacts participants of different demographic and professional backgrounds or those who participate in different training modalities. Using a pre-post follow-up design with a comparison group, this study examined changes in participants confidence in their ability to recognize and support youth mental health needs 1 to 2 months after training. Data were collected over two years (2021-2023) from training participants (n = 480) and comparable non-participants (n = 51). The authors examined whether changes in confidence varied by participant race/ethnicity, professional role in the education or mental health fields, and training modality (online versus hybrid). Training participants confidence in supporting youth mental health increased significantly compared to non-participants. Although the training was effective for all participants, those with less mental health experience benefited more, consistent with previous research. While both in-person and hybrid training were effective, in-person training participants reported slightly higher confidence scores than virtual at follow-up. Study findings suggest that educational and social service organizations should offer this training to their staff and community members who interact with youth, prioritizing participants with less prior mental health training and delivering training through an in-person training modality when possible. However, additional research is needed to explore how aspects of in-person training, such as trainer characteristics and group dynamics, impact outcomes

Developing National Standardized Performance Measures for School‐Based Health Centers: The National Quality Initiative*

BackgroundDespite extensive literature on school-based health center (SBHC) characteristics and outcomes, their quality of care has not been examined nationally. Standardized quality metrics can inform health care delivery and improvement.MethodsSBHC national performance measures (NPMs) were developed by reviewing measures from national child health quality initiatives and engaging stakeholders in a consensus-building process. NPMs were pilot-tested with 73 SBHCs and SBHCs nationally subsequently reported data.ResultsFive NPMs were selected including the percentage of clients annually who received at least one: (1) well-child visit, whether administered in the SBHC or elsewhere; (2) risk assessment; (3) body mass index screen with nutrition and physical activity counseling; and, if age-appropriate, (4) depression screening with follow-up treatment plan; and (5) chlamydia screening among sexually active clients. SBHCs experienced challenges with reporting during pilot-testing, particularly related to extracting data from electronic health records, and identified strategies to address challenges. Approximately 20% of SBHCs nationally voluntarily reported data during the initial year.Implications for school healthStandardized performance measures can help SBHCs monitor and improve care delivery and demonstrate effectiveness compared to other child health delivery systems.ConclusionOngoing data collection will help examine whether measure adoption drives quality improvement for SBHCs nationwide

Patent: Methods of Treating FGF21-Associated Disorders

The invention relates to the identification of new polypeptide and protein variants of fibroblast growth factor 21 (FGF21) that have improved pharmaceutical properties. Also disclosed are methods for treating FGF21-associated disorders, includ ing metabolic conditions

Patent: Dual Function Proteins for Treating Metabolic Disorders

The present invention relates to new proteins comprising fibroblast growth factor 2 1 (FGF21 ) and other metabolic regulators known to improve metabolic profiles in subjects to whom they are administered

Structural analysis of covalent peptide dimers, bis(pyridine-2-carboxamidonetropsin)(CH_2)_(3-6), in complex with 5'-TGACT-3' sites by two-dimensional NMR

The peptide pyridine-2-carboxamidonetropsin (2-PyN) binds specifically in the minor groove of 5'-(A,T)G(A,T)C(A,T)-3' sequences as a side-by-side antiparallel dimer. Tethering two 2-PyN ligands through the nitrogens of the central pyrrole rings with propyl, butyl, pentyl and hexyl linkers affords covalent peptide dimers, bis(pyridine-2-carboxamide-netropsin)(CH_2)_(3-6), which bind in the minor groove of DNA with increased binding affinities and improved sequence specificities. Two-dimensional NMR studies of the complexes formed upon binding of these covalent peptide dimers to an oligonucleotide containing a 5'-TGACT-3' site reveal that the dimeric peptides bind as intramolecular dimers with nearly identical geometry and peptide-DNA contacts as in the (2-PyN)_2•5'-TGACT-3' complex

Structural and dynamic characterization of the heterodimeric and homodimeric complexes of distamycin and 1-methylimidazole-2-carboxamide-netropsin bound to the minor groove of DNA

NMR spectroscopy combined with molecular modeling was used to characterize a heterodimeric complex with Dst and 2-ImN bound in the minor groove of d(GCCTAACAAGG)•d(CCTTGTTAGGC) (1:1:1 2-ImN•Dst•DNA complex). The imidazole-pyrrole-pyrrole ligand 2-ImN spans 5'-GTTA-3' of the TAACA•TGTTA binding site with the imidazole nitrogen specifically recognizing the guanine amino group. The Dst ligand lies along the 5'-AACA-3' sequence and complements the 2-ImN ligand in the formation of the antiparallel side-by-side heterodimeric complex. Titrations of the same site with Dst or 2-ImN alone yield homodimeric complexes (2:1 ligand.DNA) of lower stability than the 1:1:1 2-ImN•Dst•DNA complex. Dst and 2-ImN binding to d(CGCAAACTGGC)•d(GCCAGTTTGCG) was also investigated. The 1:1:1 2-ImN•Dst•DNA complex is again the most stable complex with the AAACT•AGTTT site and is similar to the TAACA•TGTTA complex. No monomeric binding of either 2-ImN or Dst was observed to either site

NMR characterization of hairpin polyamide complexes with the minor groove of DNA

Polyamides containing N-methylimidazole (Im) and N-methylpyrrole (Py) amino acids can be combined in antiparallel side-by-side dimeric complexes for sequence-specific recognition in the minor groove of DNA. Covalently linking polyamide subunits has led to designed ligands with both increased affinity and specificity. Simple aliphatic amino acid linkers serve as internal guide residues for turn vs extended binding in a head-to-tail-linked polyamide motif. Polyamides of sequence composition ImPyPy-X-PyPyPy containing linkers of incremental length (X = 3-aminopropionic acid (β), 4-aminobutyric acid (γ), or 5-aminovaleric acid (δ)) in complex with an undecamer DNA duplex containing a 5'-(A,T)G(A,T)(3)-3' target site were structurally characterized using NMR spectroscopy. Previous quantitative DNase I footprinting studies identified gamma as the highest affinity of these "turn" linkers. NMR titrations and 2D NOESY data combined with restrained molecular modeling reveal that polyamides with β, γ, and δ linkers all may adopt a hairpin structure. Modeling supports the idea that the linkers in the βand δcomplexes adopt an energetically less favorable turn geometry than the γlinker and confirms that the three-carbon γ linker is sufficient and optimal for the hairpin conformation