A reduction in brain mass and cortical volume seen in the adult <i>Cln6^nclf</i> mouse.

<p>Brain mass was assessed in the <i>Cln6^nclf</i>, as a decrease in brain mass is often seen in vLINCL patients. (<b>A</b>) Brain mass was reduced beginning at 5 months when compared to age matched controls. (<b>B</b>) Hippocampal and cerebral cortex volume were further assessed revealing a decrease in cortical volume at 9 months in the <i>Cln6^nclf</i> mouse. (<b>C</b>) No difference in mean body weight is seen between adult WT and <i>Cln6^nclf</i> mice up to 12 months of age. [Mean +/− SEM, <i>n</i> = 3 (**<i>p</i>≤0.01, ***<i>p</i>≤0.0001) for brain mass (A) and cortical volume measurements; <i>n</i> = 7–12 for body weight measurements]].</p

Learning and memory deficits are associated with mutation in <i>Cln6</i>.

<p>Following a period of habituation and training, memory and learning performance were tested in a radial arm maze task. <b>(A–B)</b> In an assay of memory, <i>Cln6^nclf</i> mice displayed a significant increase in the latency (time in seconds) to complete an 8-arm radial maze task during a 5-trial session. <b>(C–D)</b> Deficits in the mean amount of time required to complete the task <b>(C)</b> and the mean total distance traveled (in meters) to complete the correct maze sequence <b>(D)</b> occurred in <i>Cln6^nclfmice</i> when the maze task was altered 24-hours later to assay learning. [Mean +/− SEM, <i>n</i> = 6–9 (**<i>p</i>≤0.01, ***<i>p</i>≤0.0001)].</p

Loss of specific subpopulations of interneurons seen within various regions of the hippocampus.

<p>Loss of interneurons is a characteristic often seen in the hippocampus of the NCLs. We divided the hippocampus into defined regions and stained sections for specific subpopulations of interneurons and subsequently counted the subpopulations within the sections. (<b>A</b>) PV+ interneurons decreased in the dentate gyrus (DG) at 9 months but remain unchanged in other areas where PV+ interneurons were present. (<b>B</b>) SOM+ interneurons were first decreased at 5 months in the hilus and stratum oriens with further decline observed at 9 months. Decline of SOM+ interneurons was also seen at 9 months in the stratum radiatum as well as the combined CA fields of the hippocampus. (<b>C–D</b>) A significant reduction in the number of CB+ interneurons was observed within the stratum oriens at 9 months when compared to WT. [Mean +/− SEM, <i>n</i> = 3 (*<i>p</i>≤0.05, **<i>p</i>≤0.01, ***p≤0.001, ****<i>p</i>≤0.0001)].</p

Decreased dendritic spine density in the mature cortex of <i>Cln6^nclf</i> mice.

<p>Golgi impregnation was used to label dendritic spines on the primary dendrite of excitatory cortical projection neurons in age-matched 2 month controls (<b>A</b>) and <i>Cln6^nclf</i> (<b>B</b>) mice. <b>(C)</b> The <i>Cln6^nclf</i> mutant mouse excitatory pyramidal neurons have reduced spine density. [Mean spine density +/− SEM, <i>n</i> = 40 neurons counted per genotype (***<i>p</i>≤0.0001)].</p

Decreased motor coordination deficits in <i>Cln6^nclf</i> mice.

<p><b>(A)</b> Rotarod testing was performed on postnatal day 14, 28, 90, and 270 old WT and <i>Cln6^nclf</i> mice. Data are plotted as average latency to fall from the rotating rod during a 240 second trial period (3 trials per mouse per time point). <i>Cln6^nclf</i> mice had a significant reduction in their ability to remain on the rod as it accelerated, starting at P90 and continuing at P270. <b>(B–C)</b> At six months of age, no difference was noted in additional motor performance measures including the time required to descend in a pole climb test <b>(B)</b> or the mean distance traveled (in meters) over a 15-min test period in an open field activity test <b>(C)</b>. [Mean +/− SEM, <i>n</i> = 6–9 mice per group (**<i>p</i>≤0.01, ***<i>p</i>≤0.0001)].</p

Retinal degeneration and vision loss in the <i>Cln6^nclf</i> mouse.

<p>Cell loss and structural degenerative changes occur in the retina of <i>Cln6^nclf</i> mice. (<b>A</b>) Comparison of gross morphological changes over time in retina of <i>Cln6^nclf</i> mice and their respective age-matched WT controls was done to determine mechanism of degeneration. (<b>B</b>) Micrographs (4X) show a section of one retinal hemisphere. (<b>C</b>) At P0 all layers are present and of equal thickness. By 3 months of age the <i>Cln6^nclf</i> retina has begun to narrow and shows a distinct loss of the rods and cones while the overall cytoarchitecture remains intact. By 9 months the rods and cones are nearly absent and the outer plexiform layer is virtually nonexistent with the merging of a much thinned outer and inner nuclear layers. Additionally, there is a distinct narrowing of the inner plexiform layer. [RC-Rode/Cone layer; ONL-Outer nuclear layer; OPL-Outer plexiform later; INL-Inner nuclear layer; IPL-Inner plexiform layer; GCL-Ganglion cell layer]. (D) At 8 months of age, <i>Cln6^nclf</i> mice displayed a significant reduction in visual acuity in a visual cliff assay. Mutant mice were unable to distinguish between a “safe” region of the visual cliff box versus the “unsafe” cliffed portion, spending equal time between the two regions. [Mean (in seconds) +/- SEM, <i>n</i> = 6–9 mice per group (**<i>p</i>≤0.01)].</p

Upregulation of microglial marker CD68 in the thalamus and cerebral cortex of <i>Cln6^nclf</i> mice.

<p>Quantitative thresholding analysis in <i>Cln6^nclf</i> mice was compared to age matched WT, revealing a significant increase in the expression of the microglia marker CD68 in the VPM/VPL (<b>A</b>), M1 (<b>C</b>), S1BF (<b>B</b>), and V1 (<b>D</b>) regions in the <i>Cln6^nclf</i> mouse over WT. [Mean% immunoreactivity +/− SEM, <i>n</i> = 3 (*<i>p</i>≤0.05, **<i>p</i>≤0.01, ***p≤0.001, ****<i>p</i>≤0.0001)].</p

Cortical atrophy limited in <i>Cln6^nclf</i> mice.

<p>Cortical thickness was evaluated in age matched WT and <i>Cln6^nclf</i> mice. (<b>A</b>) In the S1BF, thinning begins to appear at 6 months. (<b>B</b>) Thinning in the M1 region is seen at 11 months. (<b>C</b>) Cortical thickness was unchanged in the LEnt (<b>C</b>) region of the cerebral cortex. (D–E) The S1BF (<b>D</b>) and M1 (<b>E</b>) regions were analyzed for possible laminar specific atrophy. Thinning is seen in lamina V of the M1 region at 11 months as well as an apparent thickening in lamina IV. Thinning is seen in the S1BF region in lamina VI at 6 months. [Mean +/− SEM (<i>n</i> = 3,*<i>p</i>≤0.05, **<i>p</i>≤0.01)].</p

Upregulation of astrocyte marker GFAP in the thalamus and cerebral cortex of <i>Cln6^nclf</i> mice.

<p>(<b>A</b>) Quantitative thresholding analysis revealed a significant increase in the expression of astrocytic markers in the <i>Cln6^nclf</i> mouse at 4 months in the thalamus. (<b>B–C</b>) Elevations in astrocytic activation, marked by GFAP labeling, became apparent in the S1BF region at 4 months and in the V1 region of the cerebral cortex at 6 months in the <i>Cln6^nclf</i> mouse. [Mean% immunoreactivity +/− SEM, <i>n</i> = 3 (*<i>p</i>≤0.05, **<i>p</i>≤0.01, ***<i>p</i>≤0.0001)].</p

Association of Medication Adherence with HbA1c Control among American Indian Adults with Type 2 Diabetes Using Tribal Health Services

   OBJECTIVE To examine HbA1c levels and adherence to oral glucose-lowering medication and their association with future HbA1c levels among American Indian adults with type 2 diabetes (T2D) receiving medication at no-cost from a tribal healthcare system.  RESEARCH DESIGN AND METHODS Tribal citizens with T2D and who used Choctaw Nation Health Services Authority (CNHSA) and Pharmacies and had HbA1c data during 2017-2018 were included in this study. Medication adherence (proportion of days covered [PDC] ≥ 0.80) was calculated using 2017 CNHSA electronic health record data.  RESULTS Of the 74,000 tribal citizens living on tribal lands, 4,560 were eligible. 32% had HbA1c at or below target (≤ 7%); 36% were above target (> 7% to ≤ 9%); 32% were uncontrolled (> 9%) in 2017. Percentage of patients with PDC ≥.80 was 66% for Biguanides, 72% for Sulfonylureas, 75% for DPP-4 inhibitors, and 83% for SGLT-2 inhibitors. The proportion of patients with HbA1c at or below target increased slightly from 32% in 2017 to 42% in 2018. Higher average PDC in 2017 was associated with lower HbA1c levels in 2018 (β=-1.143, p CONCLUSION Medication adherence was higher than found in previous studies that used self-report methods in American Indian populations, though a smaller proportion of patients had an HbA1c at or below target relative to US adults with T2D. Mediation adherence was associated with improved HbA1c level for most oral glucose-lowering medication classes. Future studies of American Indians should use both longitudinal prescription data from both electronic health record and pharmacy refill data. </p