Description of Specialty Practice-10 Years Onward: The Changes in Geriatric Physical Therapy.

BACKGROUND: An analysis of practice is conducted by the American Board of Physical Therapy Specialties (ABPTS) every 10 years to revalidate, update, and revise the description of specialty practice (DSP) for each specialty. The Geriatric Specialty Council recently conducted an analysis of practice and revised its content consistent with established procedures by the ABPTS. PURPOSE: The purpose of this article is threefold: first, to describe the process of the most recent practice analysis; second, to report revisions to the description of specialty practice based on the analysis of practice; and third, to identify elements of practice that define current specialist practice in geriatric physical therapy. METHODS: A 10-member committee of subject matter experts (SMEs) and a psychometric consultant developed a survey instrument addressing geriatric physical therapy specialty practice areas. The survey was initially pilot-tested and subsequently administered online to a sample of 801 board-certified geriatric clinical specialists. The consultant facilitated the consensus process to determine decision rules in selecting the final competencies describing current geriatric physical therapy specialty practice. RESULTS: A total of 372 respondents fully or partially completed the survey, resulting in a response rate of 46.4%. Based on a priori decision rules regarding survey data, consensus of the group of SMEs, and input from the ABPTS, the DSP for geriatric physical therapy specialty practice was revised. Revisions (elimination [-] of prior items and addition [+] of new items) were made in Section 1: Knowledge Areas (-8 and +6), in Section 2: Professional Roles, Responsibilities, and Values (-14 and +4), and Section 3: Practice Expectations (-53 and +28). CONCLUSION: The revised DSP will be used as the basis for the development of the examination blueprint for the specialist examination in geriatric physical therapy and the curricula for residency programs in geriatric physical therapy.

Opportunities and Challenges to Linkage to Housing in the Context of a Sexual and Reproductive Health Program for Youth Experiencing Homelessness.

Youth homelessness remains a major public health issue in the USA, and youth experiencing homelessness (YEH) are still one of the more understudied and underserved groups. Comprehensive sexual and reproductive health (SRH) programs for YEH are rare. Yet, such programs are potentially effective settings through which to link YEH with housing services. Wahine ("woman") Talk is one such comprehensive program for YEH, and is a multilevel intervention delivered out of a youth drop-in center in Honolulu, Hawai 'i. One of Wahine Talk's core components is addressing basic needs, including providing linkages to housing services. Little research exists on SRH programs' opportunities and challenges to providing linkage to housing for YEH. The current study is an exploratory study asking, "What are opportunities and challenges to linking young women experiencing homelessness to housing services through a comprehensive sexual and reproductive health program?" The study team collected in-depth qualitative data through seven focus groups and 25 individual interviews with Wahine Talk staff and youth participants aged 14-22 years. Multiple team members analyzed the data using template analysis. The analysis revealed that while comprehensive SRH programs may have some opportunities and challenges to linking YEH to housing services that are consistent with traditional housing assistance programs, there are also factors specific to SRH programs. In particular, opportunities would be SRH programs employing a housing staff person and bolstering staff-youth meetings and communication. A challenge specific to SRH programs would be prioritizing youth's reproductive justice (i.e., choice) in lieu of solely prioritizing pregnancy reductions and delays; thus, it is recommended to train staff to prioritize youth's reproductive justice. The findings highlight the importance of SRH programs having staff focused on housing, sufficient opportunities for youth and staff to communicate with each other, and staff trained to prioritize youth's reproductive justice.

Effect of Variation in Early Rehabilitation on Hospital Readmission After Hip Fracture.

OBJECTIVE: Provision of early rehabilitation services during acute hospitalization after a hip fracture is vital for improving patient outcomes. The purpose of this study was to examine the association between the amount of rehabilitation services received during the acute care stay and hospital readmission in older patients after a hip fracture. METHODS: Medicare claims data (2016-2017) for older adults admitted to acute hospitals for a hip fracture (n = 131,127) were used. Hospital-based rehabilitation (physical therapy, occupational therapy, or both) was categorized into tertiles by minutes per day as low (median = 17.5), middle (median = 30.0), and high (median = 48.8). The study outcome was risk-adjusted 7-day and 30-day all-cause hospital readmission. RESULTS: The median hospital stay was 5 days (interquartile range [IQR] = 4-6 days). The median rehabilitation minutes per day was 30 (IQR = 21-42.5 minutes), with 17 (IQR = 12.6-20.6 minutes) in the low tertile, 30 (IQR = 12.6-20.6 minutes) in the middle tertile, and 48.8 (IQR = 42.8-60.0 minutes) in the high tertile. Compared with high therapy minutes groups, those in the low and middle tertiles had higher odds of a 30-day readmission (low tertile: odds ratio [OR] = 1.11, 95% CI = 1.06-1.17; middle tertile: OR = 1.07, 95% CI = 1.02-1.12). In addition, patients who received low rehabilitation volume had higher odds of a 7-day readmission (OR = 1.20; 95% CI = 1.10-1.30) compared with high volume. CONCLUSION: Elderly patients with hip fractures who received less rehabilitation were at higher risk of readmission within 7 and 30 days. IMPACT: These findings confirm the need to update clinical guidelines in the provision of early rehabilitation services to improve patient outcomes during acute hospital stays for individuals with hip fracture. LAY SUMMARY: There is significant individual- and hospital-level variation in the amount of hospital-based rehabilitation delivered to older adults during hip fracture hospitalization. Higher intensity of hospital-based rehabilitation care was associated with a lower risk of hospital readmission within 7 and 30 days.

Mind the GAP: Purification and characterization of urea resistant GAPDH during extreme dehydration.

The African clawed frog (Xenopus laevis) withstands prolonged periods of extreme whole-body dehydration that lead to impaired blood flow, global hypoxia, and ischemic stress. During dehydration, these frogs shift from oxidative metabolism to a reliance on anaerobic glycolysis. In this study, we purified the central glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to electrophoretic homogeneity and investigated structural, kinetic, subcellular localization, and post-translational modification properties between control and 30% dehydrated X. laevis liver. GAPDH from dehydrated liver displayed a 25.4% reduction in maximal velocity and a 55.7% increase in its affinity for GAP, as compared to enzyme from hydrated frogs. Under dehydration mimicking conditions (150 mM urea and 1% PEG), GAP affinity was reduced with a Km value 53.8% higher than controls. Frog dehydration also induced a significant increase in serine phosphorylation, methylation, acetylation, beta-N-acetylglucosamination, and cysteine nitrosylation, post-translational modifications (PTMs). These modifications were bioinformatically predicted and experimentally validated to govern protein stability, enzymatic activity, and nuclear translocation, which increased during dehydration. These dehydration-responsive protein modifications, however, did not appear to affect enzymatic thermostability as GAPDH melting temperatures remained unchanged when tested with differential scanning fluorimetry. PTMs could promote extreme urea resistance in dehydrated GAPDH since the enzyme from dehydrated animals had a urea I50 of 7.3 M, while the I50 from the hydrated enzyme was 5.3 M. The physiological consequences of these dehydration-induced molecular modifications of GAPDH likely suppress GADPH glycolytic functions during the reduced circulation and global hypoxia experienced in dehydrated X. laevis.

"Having a Baby Can Wait": Experiences of a Sexual and Reproductive Health Promotion Program in the Context of Homelessness among Asian American, Native Hawaiian, and Other Pacific Islander Youth Captured Through PhotoVoice.

Nearly half of female youth experiencing homelessness (YEH) become pregnant due to myriad individual, family, community, and structural factors. In response, our team developed and tested Wahine ("woman") Talk, a multilevel, trauma-informed sexual and reproductive health intervention created with and for female YEH aged 14 to 22. After Wahine Talk, youth were invited to participate in a participatory action PhotoVoice project regarding experiences of the program, waiting to start or expand their families, and homelessness. Photographs were taken and captioned by youth, and then further examined through Thematic Analysis. Final project themes include (a) Youth-Driven Birth Timing Decisions; (b) A Sense of Place: Finding Safe Spaces; and (c) Glimpsing Hope. Because YEH live under society's radar, it is critical to understand their experiences from their own perspectives to improve interventions at multiple levels. Implications for meeting the needs of YEH in the areas of reproductive justice, financial stability, and affordable housing are discussed.

Differential protein phosphorylation is responsible for hypoxia-induced regulation of the Akt/mTOR pathway in naked mole rats.

Naked mole rats (NMRs, Heterocephalus glaber) are among the most hypoxia-tolerant mammals known. They can reduce their metabolic rate (>85%) under severe hypoxia, remain moderately active and recover with no obvious signs of damage. Hence, NMRs are an excellent model for studying mammalian hypoxia tolerance. The current study characterized the involvement of posttranslational modifications in regulating the Akt/mTOR pathway that regulates protein synthesis, and the responses of key ribosomal proteins in order to assess tissue-specific responses to 4 h exposure to 7% O2 (compared to controls at 21% O2). Results showed a tissue-specific regulation of the Akt/mTOR pathway via differential phosphorylation. Relative amounts of p-TSC(S939) in brain and of p-TSC(S939), p-Akt(473) and p-PTEN(S380) in liver increased under hypoxia, whereas levels of IGF1R(Y1135/1136) in liver decreased. In skeletal muscle, levels of p-Akt(S473) and p-PTEN(S380) decreased during hypoxia, whereas lungs showed an increase in p-mTOR(S2884) content but a decrease in p-RPS6(S235-236) under the same conditions. Analysis of the phosphorylation states of ribosomal proteins revealed increases in p-4E-BP1(T37/46) content in brain and lungs under hypoxia, as well as a rise in total 4E-BP1 protein level in liver. Phosphorylated eIF-4B(S422) content also increased in liver while levels of p-eIF-2α(S51), and eIF-4E(S209) decreased during hypoxia in liver. Overall, hypoxia altered the Akt/mTOR pathway, which correlated with a general decrease in activity of the ribosomal protein biosynthesis machinery in muscle, lung, and brain of NMRs. However, the increase in eIF-4B in liver suggests the potential promotion of cap-independent mRNA translation mechanism operating under hypoxic stress.

The heart of a hibernator: EGFR and MAPK signaling in cardiac muscle during the hibernation of thirteen-lined ground squirrels, Ictidomys tridecemlineatus.

BACKGROUND: Thirteen-lined ground squirrels (Ictidomys tridecemlineatus) experience dramatic changes in physiological and molecular parameters during winter hibernation. Notably, these animals experience reduced blood circulation during torpor, which can put numerous stresses on their hearts. The present study evaluates the role played by the epidermal growth factor receptor (EGFR) in signal transduction during hibernation at low body temperature to evaluate signaling mechanisms. By investigating the regulation of intracellular mitogen activated protein kinase (MAPK) pathway responses, anti-apoptosis signals, downstream transcription factors, and heat shock proteins in cardiac muscle we aim to determine the correlation between upstream tyrosine phosphorylation events and downstream outcomes. METHODS: Protein abundance of phosphorylated EGFR, MAPKs and downstream effector proteins were quantified using immunoblotting and Luminex® multiplex assays. RESULTS: Monitoring five time points over the torpor/arousal cycle, EGFR phosphorylation on T654, Y1068, Y1086 was found to increase significantly compared with euthermic control values particularly during the arousal process from torpor, whereas phosphorylation at Y1045 was reduced during torpor. Phosphorylation of intracellular MAPK targets (p-ERK 1/2, p-JNK, p-p38) also increased strongly during the early arousal stage with p-p38 levels also rising during prolonged torpor. However, of downstream MAPK effector kinases that were measured, only p-Elk-1 levels changed showing a decrease during interbout arousal (IA). Apoptosis markers revealed a strong reduction of the pro-apoptotic p-BAD protein during entrance into torpor that remained suppressed through torpor and IA. However, active caspase-9 protein rose strongly during IA. Levels of p-AKT were suppressed during the transition phases into and out of torpor. Of four heat shock proteins assessed, only HSP27 protein levels changed significantly (a 40% decrease) during torpor. CONCLUSION: We show evidence of EGFR phosphorylation correlating to activation of MAPK signaling and downstream p-ELK1 suppression during hibernation. We also demonstrate a reduction in p-BAD mediated pro-apoptotic signaling during hibernation with active caspase-9 protein levels increasing only during IA. I. tridecemlineatus has natural mechanisms of tissue protection during hibernation that is largely due to cellular regulation through phosphorylation-mediated signaling cascade. We identify a possible link between EGFR and MAPK signaling via p-ERK, p-p38, and p-JNK in the cardiac muscle of these hibernating mammals that correlates with an apparent reduction in caspase-9 apoptotic signaling. This reveals a piece of the mechanism behind how these mammals are resilient to cardiac stresses during hibernation that would otherwise be damaging.

Characterization of ice recrystallization inhibition activity in the novel freeze-responsive protein Fr10 from freeze-tolerant wood frogs, Rana sylvatica.

Fr10 is a secreted freeze-responsive protein found in the wood frog (Rana sylvatica). This protein has gained notable research attention for its highly dynamic expression in response to seasonal freezing stress, while its over-expression has been documented to enhance freeze tolerance in cold-susceptible cultured cells. This study further characterizes the properties of this novel protein with regards to thermal stability and ice recrystallization inhibition (i.e. IRI) activity. Thermal stability was assessed using differential scanning fluorimetry, with an experimental Tm value of 50.8 ±â€¯0.1 °C. Potential IRI activity of Fr10 was evaluated using a recently developed nanoparticle-based colorimetric assay, where Fr10 displayed the ability to prevent freeze-induced aggregation of gold nanoparticles. Based upon this assay, Fr10 protein appeared to have a low level of IRI activity and it was therefore predicted that one of Fr10's biological functions may be to inhibit ice crystal growth via recrystallization. A SPLAT cooling assay was then employed to directly characterize the IRI properties of Fr10 and provide further insight into this hypothesis. In the presence of 30 µM of Fr10, a 40% reduction in the mean grain size of ice crystals relative to the control samples was observed, thus introducing the possibility of Fr10 to inhibit ice recrystallization. Collectively, the results from this study provide new insight into the potential of further exploring the potential of this vertebrate freeze-responsive protein in cryoprotection.

Purification and characterization of a urea sensitive lactate dehydrogenase from skeletal muscle of the African clawed frog, Xenopus laevis.

The African clawed frog, Xenopus laevis endures whole body dehydration which can increase its reliance on anaerobic glycolysis for energy production. This makes the regulation of the terminal enzyme of glycolysis, lactate dehydrogenase (LDH), crucial to stress survival. We investigated the enzymatic properties and posttranslational modification state of purified LDH from the skeletal muscle of control and dehydrated (30% total body water loss) X. laevis. LDH from the muscle of dehydrated frogs showed a 93% reduction in phosphorylation on threonine residues and an 80% reduction of protein nitrosylation. LDH from dehydrated muscle also showed a 74% lower Vmax in the pyruvate oxidizing direction and a 78% decrease in Vmax in the lactate reducing direction along with a 33% lower Km for pyruvate and a 40% higher Km for lactate. In the presence of higher levels of urea and molecular crowding by polyethylene glycol, used to mimic conditions in the cells of dehydrated animals, the Km values of control and dehydrated LDH demonstrated opposite responses. In the pyruvate oxidizing direction, control muscle LDH was unaffected by these additives, whereas the affinity for pyruvate dropped further for LDH from dehydrated muscle. The opposite effect was more pronounced in the lactate reducing direction as control LDH showed an increased affinity for lactate, whereas LDH from dehydrated animals showed a further reduction in affinity. The physiological consequences of dehydration-induced LDH regulation appear to poise the enzyme towards lactate production when urea levels are high and lactate catabolism when urea levels are low, perhaps helping to maintain glycolysis under dehydrating conditions whilst providing for the ability to recycle lactate upon rehydration.

-Characterization of pyruvate kinase from the anoxia tolerant turtle, Trachemys scripta elegans: a potential role for enzyme methylation during metabolic rate depression.

BACKGROUND: Pyruvate kinase (PK) is responsible for the final reaction in glycolysis. As PK is a glycolytic control point, the analysis of PK posttranslational modifications (PTM) and kinetic changes reveals a key piece of the reorganization of energy metabolism in an anoxia tolerant vertebrate. METHODS: To explore PK regulation, the enzyme was isolated from red skeletal muscle and liver of aerobic and 20-hr anoxia-exposed red eared-slider turtles (Trachemys scripta elegans). Kinetic analysis and immunoblotting were used to assess enzyme function and the corresponding covalent modifications to the enzymes structure during anoxia. RESULTS: Both muscle and liver isoforms showed decreased affinity for phosphoenolpyruvate substrate during anoxia, and muscle PK also had a lower affinity for ADP. I50 values for the inhibitors ATP and lactate were lower for PK from both tissues after anoxic exposure while I50 L-alanine was only reduced in the liver. Both isozymes showed significant increases in threonine phosphorylation (by 42% in muscle and 60% in liver) and lysine methylation (by 43% in muscle and 70% in liver) during anoxia which have been linked to suppression of PK activity in other organisms. Liver PK also showed a 26% decrease in tyrosine phosphorylation under anoxia. DISCUSSION: Anoxia responsive changes in turtle muscle and liver PK coordinate with an overall reduced activity state. This reduced affinity for the forward glycolytic reaction is likely a key component of the overall metabolic rate depression that supports long term survival in anoxia tolerant turtles. The coinciding methyl- and phospho- PTM alterations present the mechanism for tissue specific enzyme modification during anoxia.

The roles of hippocampal microRNAs in response to acute postnatal exposure to di(2-ethylhexyl) phthalate in female and male rats.

Previous studies have shown that di(2-ethylhexyl) phthalate (DEHP) exposure impairs the normal development of pre- and post-synaptic elements of the male, but not female, rat hippocampus. While males seem to be vulnerable to the neurodevelopmental deficits resulting from DEHP exposure, females appear to show a protective response. The purpose of the present study was to characterize hippocampal microRNAs in female and male rats exposed to DEHP to assess whether any patterns emerged that would be consistent with vulnerability in males and resilience in females. Male and female rats were treated with 0, 1, 10, or 20mg/kg of DEHP by intraperitoneal injections from postnatal day 16 (PND16) - PND22 and brains were removed and flash frozen on PND78. A group of 85 microRNAs which have been previously shown to play a role in the development and maintenance of hippocampal neurons was assessed with RT-qPCR. In response to DEHP exposure, there were 19 microRNAs that increased in females and 52 that decreased in males. The strongest microRNA response in females occurred in conjunction with the 10mg/kg of DEHP dose, whereas suppression of microRNAs in males appeared to be dose-dependent. Select hippocampal microRNAs (such as miR-132-3p and miR-191-5p), previously shown to regulate dendrite morphology, were modulated by DEHP exposure in this study. The results suggest that DEHP exposure has the potential to regulate microRNAs in a sex-specific manner which may interfere with proper hippocampal development in males and preserve hippocampal development in females.

Purification and properties of glycerol-3-phosphate dehydrogenase from the liver of the hibernating ground squirrel, Urocitellus richardsonii.

Cytosolic glycerol-3-phosphate dehydrogenase (G3PDH, EC 1.1.1.8) is an important branch point enzyme connecting lipid metabolism and carbohydrate metabolism. We investigated the dynamic nature of G3PDH by purifying the enzyme from the liver of Richardson's ground squirrel (Urocitellus richardsonii), a hibernating species, and analyzing its structural and functional changes during hibernation. Kinetic parameters of purified G3PDH from ground squirrel liver were characterized at 37, 22 and 5°C and compared between euthermic and hibernating states. Relative to euthermic liver G3PDH, hibernator liver G3PDH had a decreased affinity for its substrate, glycerol-3-phosphate (G3P), at 37°C and 22°C. However, at 5°C, there was a significant increase in the affinity for G3P in the hibernating form of the enzyme, relative to the euthermic form. Furthermore, the structure of G3PDH in the species' hibernating state showed greater thermal stability compared to its structure in the euthermic state. Western blot analysis revealed greater tyrosine phosphorylation in hibernator G3PDH as compared to euthermic G3PDH. In addition, using the protein sequence of the hibernating thirteen-lined ground squirrel (Ictidomys tridecemlineatus) and bioinformatics tools, a three-dimensional model of G3PDH was built to identify the potential phosphorylation site (83Tyr) responsible for the differential phosphorylation between euthermic and hibernator G3PDH. The structural and functional changes in G3PDH support the enzyme's function at a low core body temperature experienced during the species hibernating season.

Native denaturation differential scanning fluorimetry: Determining the effect of urea using a quantitative real-time thermocycler.

The effect of protein stability on kinetic function is monitored with many techniques that often require large amounts of expensive substrates and specialized equipment not universally available. We present differential scanning fluorimetry (DSF), a simple high-throughput assay performed in real-time thermocyclers, as a technique for analysis of protein unfolding. Furthermore, we demonstrate a correlation between the half-maximal rate of protein unfolding (Knd), and protein unfolding by urea (I50). This demonstrates that DSF methods can determine the structural stability of an enzyme's active site and can compare the relative structural stability of homologous enzymes with a high degree of sequence similarity.

Post-translational Regulation of Hexokinase Function and Protein Stability in the Aestivating Frog Xenopus laevis.

Xenopus laevis endure substantial dehydration which can impose hypoxic stress due to impaired blood flow. Tissues may increase reliance on anaerobic glycolysis for energy production making the regulation of hexokinase (HK) important. We investigated the enzymatic properties and phosphorylation state of purified HK from the muscle of control and dehydrated (30% total body water lost) frogs. Bioinformatic tools were also applied to analyze the structural implication of HK phosphorylation in silico. HK from the muscle of dehydrated frogs showed a significantly higher Vmax (3.4-fold) and Km for glucose (2.4-fold) compared with control HK but the Km for ATP was unaltered. HK from dehydrated frogs also showed greater phosphoserine content (20% increase) and lower phosphothreonine (22% decrease) content compared to control HK. Control HK had a higher melting temperature (Tm = 61.9 °C) than from dehydrated (Tm = 54.2 °C) frogs when thermostability was tested using differential scanning fluorimetry. In silico phosphorylation of a Xenopus HK caused alterations in active site binding, corroborating phosphorylation as the probable mechanism for kinetic regulation. Physiological consequences of dehydration-induced HK phosphorylation appear to facilitate glycolytic metabolism in hypoxic situations. Augmented HK function increases the ability of Xenopus to overcome compromised oxidative phosphorylation associated with ischemia during dehydration.

Regulation of the Rana sylvatica brevinin-1SY antimicrobial peptide during development and in dorsal and ventral skin in response to freezing, anoxia and dehydration.

Brevinin-1SY is the only described antimicrobial peptide (AMP) of Rana sylvatica. As AMPs are important innate immune molecules that inhibit microbes, this study examined brevinin-1SY regulation during development and in adult frogs in response to environmental stress. The brevinin-1SY nucleotide sequence was identified and used for protein modeling. Brevinin-1SY was predicted to be an amphipathic, hydrophobic, alpha helical peptide that inserts into a lipid bilayer. Brevinin-1SY transcripts were detected in tadpoles and were significantly increased during the later stages of development. Effects of environmental stress (24 h anoxia, 40% dehydration or 24 h frozen) on the mRNA levels of brevinin-1SY in the dorsal and ventral skin were examined. The brevinin-1SY mRNA levels were increased in dorsal and ventral skin of dehydrated frogs, and in ventral skin of anoxic frogs, compared with controls (non-stressed). Brevinin-1SY protein levels in peptide extracts of dorsal skin showed a similar, but not significant, trend to that of brevinin-1SY mRNA levels. Antimicrobial activity of skin extracts from control and stressed animals were assessed for Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae, Botrytis cinerea, Rhizopus stolonifer and Pythium sulcatum using disk diffusion assays. Peptide extracts of dorsal skin from anoxic, frozen and dehydrated animals showed significantly higher inhibition of E. coli and P. sulcatum than from control animals. In ventral skin peptide extracts, significant growth inhibition was observed in frozen animals for E. coli and P. sulcatum, and in anoxic animals for B. cinerea, compared with controls. Environmental regulation of brevinin-1SY may have important implications for defense against pathogens.