Targeting Molecular Mechanisms of Obesity- and Type 2 Diabetes Mellitus-Induced Skeletal Muscle Atrophy with Nerve Growth Factor.

Skeletal muscle plays a critical role in metabolic diseases, such as obesity and type 2 diabetes mellitus (T2DM). Muscle atrophy, characterized by a decrease in muscle mass and function, occurs due to an imbalance between the rates of muscle protein synthesis and degradation. This study aimed to investigate the molecular mechanisms that lead to muscle atrophy in obese and T2DM mouse models. Additionally, the effect of nerve growth factor (NGF) on the protein synthesis and degradation pathways was examined. Male mice were divided into three groups: a control group that was fed a standard chow diet, and two experimental groups that were fed a Western diet. After 8 weeks, the diabetic group was injected with streptozotocin to induce T2DM. Each group was then further divided into NGF-treated or non-treated control group. In the gastrocnemius muscles of the Western diet group, increased expressions of myostatin, autophagy markers, and ubiquitin ligases were observed. Skeletal muscle tissue morphology indicated signs of muscle atrophy in both obese and diabetic mice. The NGF-treated group showed a prominent decrease in the protein levels of myostatin and autophagy markers. Furthermore, the NGF-treated group showed an increased Cyclin D1 level. Western diet-induced obesity and T2DM may be linked to muscle atrophy through upregulation of myostatin and subsequent increase in the ubiquitin and autophagy systems. Moreover, NGF treatment may improve muscle protein synthesis and cell cycling.

Prevalence and Mechanisms of Skeletal Muscle Atrophy in Metabolic Conditions.

Skeletal muscle atrophy is prevalent in a myriad of pathological conditions, such as diabetes, denervation, long-term immobility, malnutrition, sarcopenia, obesity, Alzheimer's disease, and cachexia. This is a critically important topic that has significance in the health of the current society, particularly older adults. The most damaging effect of muscle atrophy is the decreased quality of life from functional disability, increased risk of fractures, decreased basal metabolic rate, and reduced bone mineral density. Most skeletal muscle in humans contains slow oxidative, fast oxidative, and fast glycolytic muscle fiber types. Depending on the pathological condition, either oxidative or glycolytic muscle type may be affected to a greater extent. This review article discusses the prevalence of skeletal muscle atrophy and several mechanisms, with an emphasis on high-fat, high-sugar diet patterns, obesity, and diabetes, but including other conditions such as sarcopenia, Alzheimer's disease, cancer cachexia, and heart failure.

Endometrial cancer rate in Hormone replacement therapy users with postmenopausal bleeding: Retrospective cohort study.

OBJECTIVE: To establish the endometrial cancer detection rate in women using hormone replacement therapy presenting with postmenopausal bleeding. STUDY DESIGN: Retrospective cohort study. Setting and populationRapid access gynaecology clinic at a tertiary hospital. Women aged under 60 years referred with postmenopausal bleeding. METHODS: Retrospective study of referrals received between 1 January 2019 and 31 December 2020 including Hormone replacement therapy (HRT) use and histological diagnosis. MAIN OUTCOME MEASURES: Histological diagnosis of endometrial cancer, borderline ovarian tumour or endometrial intraepithelial neoplasia. STATISTICAL ANALYSIS: Chi squared test. RESULTS: 1363 women were included. 214 women were using HRT when they experienced PMB and only one of these had endometrial cancer at histology (cancer detection rate 0.47%). 25 of the 1124 women who were not using HRT were diagnosed with endometrial cancer on histology (cancer detection rate 2.18%). Chi squared statistical analysis confirmed this was statistically significant (p value .0156). CONCLUSIONS: The endometrial cancer detection rate in women aged under 60 years using HRT with PMB is very low. Referral on a two-week wait pathway for suspected cancer diagnosis induces stress and anxiety for the woman and may lead to more invasive initial investigation even though other diagnoses are far more likely. Women aged under 60 years with postmenopausal bleeding that have either commenced HRT or had a change to their preparation within the last 6 months should be seen on a less urgent referral pathway if necessary given the very low probability of endometrial cancer.

Genistein: A focus on several neurodegenerative diseases.

Neurodegenerative diseases are caused by the progressive loss of function or structure of nerve cells in the central nervous system. The most common neurodegenerative diseases include Alzheimer's disease, Huntington's disease, motor neuron disease, and Parkinson's disease. Although the physical or mental symptoms of neurodegenerative disease may be relieved by various treatment combinations, there are currently no strategies to directly slow or prevent neurodegeneration. Given the demographic evidence of a rapidly growing aging population and the associated prevalence of these common neurodegenerative diseases, it is paramount to develop safe and effective ways to protect against neurodegenerative diseases. Most neurodegenerative diseases share some common etiologies such as oxidative stress, neuroinflammation, and mitochondrial dysfunction. Genistein is an isoflavone found in soy products that have been shown to exhibit antioxidant, anti-inflammation, and estrogenic properties. Increasing evidence indicates the protective potential of genistein in neurodegenerative disorders. In this review, we aim to provide an overview of the role that genistein plays in delaying the development of neurodegenerative disease. PRACTICAL APPLICATIONS: Genistein is a naturally occurring isoflavone found mainly in soybean, but also green peas, legumes, and peanuts. Genistein is found to pass through the blood-brain barrier and possess a neuroprotective effect. In this review, we discuss studies in support of these actions and the underlying biological mechanisms. Together, these data indicate that genistein may hold neuroprotective effects in either delaying the onset or relieving the symptoms of neurodegenerative disease.

Mitochondrial dysfunction and beneficial effects of mitochondria-targeted small peptide SS-31 in Diabetes Mellitus and Alzheimer's disease.

Diabetes and Alzheimer's disease are common chronic illnesses in the United States and lack clearly demonstrated therapeutics. Mitochondria, the "powerhouse of the cell", is involved in the homeostatic regulation of glucose, energy, and reduction/oxidation reactions. The mitochondria has been associated with the etiology of metabolic and neurological disorders through a dysfunction of regulation of reactive oxygen species. Mitochondria-targeted chemicals, such as the Szeto-Schiller-31 peptide, have advanced therapeutic potential through the inhibition of oxidative stress and the restoration of normal mitochondrial function as compared to traditional antioxidants, such as vitamin E. In this article, we summarize the pathophysiological relevance of the mitochondria and the beneficial effects of Szeto-Schiller-31 peptide in the treatment of Diabetes and Alzheimer's disease.

iPSC-derived NK cells maintain high cytotoxicity and enhance in vivo tumor control in concert with T cells and anti-PD-1 therapy.

The development of immunotherapeutic monoclonal antibodies targeting checkpoint inhibitory receptors, such as programmed cell death 1 (PD-1), or their ligands, such as PD-L1, has transformed the oncology landscape. However, durable tumor regression is limited to a minority of patients. Therefore, combining immunotherapies with those targeting checkpoint inhibitory receptors is a promising strategy to bolster antitumor responses and improve response rates. Natural killer (NK) cells have the potential to augment checkpoint inhibition therapies, such as PD-L1/PD-1 blockade, because NK cells mediate both direct tumor lysis and T cell activation and recruitment. However, sourcing donor-derived NK cells for adoptive cell therapy has been limited by both cell number and quality. Thus, we developed a robust and efficient manufacturing system for the differentiation and expansion of high-quality NK cells derived from induced pluripotent stem cells (iPSCs). iPSC-derived NK (iNK) cells produced inflammatory cytokines and exerted strong cytotoxicity against an array of hematologic and solid tumors. Furthermore, we showed that iNK cells recruit T cells and cooperate with T cells and anti-PD-1 antibody, further enhancing inflammatory cytokine production and tumor lysis. Because the iNK cell derivation process uses a renewable starting material and enables the manufacturing of large numbers of doses from a single manufacture, iNK cells represent an "off-the-shelf" source of cells for immunotherapy with the capacity to target tumors and engage the adaptive arm of the immune system to make a "cold" tumor "hot" by promoting the influx of activated T cells to augment checkpoint inhibitor therapies.

Novel Miniaturized Drug Conjugate Leverages HSP90-driven Tumor Accumulation to Overcome PI3K Inhibitor Delivery Challenges to Solid Tumors.

The PI3K pathway is considered a master regulator for cancer due to its frequent activation, making it an attractive target for pharmacologic intervention. While substantial efforts have been made to develop drugs targeting PI3K signaling, few drugs have been able to achieve the inhibition necessary for effective tumor control at tolerated doses. HSP90 is a chaperone protein that is overexpressed and activated in many tumors and as a consequence, small-molecule ligands of HSP90 are preferentially retained in tumors up to 20 times longer than in normal tissue. We hypothesize that the generation of conjugates that use a HSP90-targeting ligand and a payload such as copanlisib, may open the narrow therapeutic window of this and other PI3K inhibitors. In support of this hypothesis, we have generated a HSP90-PI3K drug conjugate, T-2143 and utilizing xenograft models, demonstrate rapid and sustained tumor accumulation of the conjugate, deep pathway inhibition, and superior efficacy than the PI3K inhibitor on its own. Selective delivery of T-2143 and the masking of the inhibitor active site was also able to mitigate a potentially dose-limiting side effect of copanlisib, hyperglycemia. These data demonstrate that by leveraging the preferential accumulation of HSP90-targeting ligands in tumors, we can selectively deliver a PI3K inhibitor leading to efficacy in multiple tumor models without hyperglycemia in mice. These data highlight a novel drug delivery strategy that allows for the potential opening of a narrow therapeutic window through specific tumor delivery of anticancer payloads and reduction of toxicity.

Novel genotype-phenotype and MRI correlations in a large cohort of patients with SPG7 mutations.

OBJECTIVE: To clinically, genetically, and radiologically characterize a large cohort of SPG7 patients. METHODS: We used data from next-generation sequencing panels for ataxias and hereditary spastic paraplegia to identify a characteristic phenotype that helped direct genetic testing for variations in SPG7. We analyzed MRI. We reviewed all published SPG7 mutations for correlations. RESULTS: We identified 42 cases with biallelic SPG7 mutations, including 7 novel mutations, including a large multi-exon deletion, representing one of the largest cohorts so far described. We identified a characteristic phenotype comprising cerebellar ataxia with prominent cerebellar dysarthria, mild lower limb spasticity, and a waddling gait, predominantly from a cohort of idiopathic ataxia. We report a rare brain MRI finding of dentate nucleus hyperintensity on T2 sequences with SPG7 mutations. We confirm that the c.1529C>T allele is frequently present in patients with long-standing British ancestry. Based on the findings of the present study and existing literature, we confirm that patients with homozygous mutations involving the M41 peptidase domain of SPG7 have a younger age at onset compared to individuals with mutations elsewhere in the gene (14 years difference, p < 0.034), whereas c.1529C>T compound heterozygous mutations are associated with a younger age at onset compared to homozygous cases (5.4 years difference, p < 0.022). CONCLUSIONS: Mutant SPG7 is common in sporadic ataxia. In patients with British ancestry, c.1529C>T allele represents the most frequent mutation. SPG7 mutations can be clinically predicted by the characteristic hybrid spastic-ataxic phenotype described above, along with T2 hyperintensity of the dentate nucleus on MRI.

Examining the Impact of This Is My Brave on Mental Illness Stigma and Willingness to Seek Help: A Pilot Study.

This Is My Brave (TIMB) is a contact-based mental illness stigma reduction program, set in theaters, meant to reduce stigma, increase beliefs about empowerment and recovery, and improve attitudes towards treatment seeking for mental health concerns. The authors conducted the first empirical evaluation of TIMB using a pre-post survey design. Approximately 481 audience members of TIMB performances were invited to complete a survey of stigmatizing attitudes towards mental illness, beliefs about recovery and empowerment, and willingness to seek treatment at pre-and post-performance. Analyses of responses from 372 participants using paired samples t-tests revealed changes in the desired direction on all variables from pre-test to post-test. Audience members experienced a decrease in stigma, improvements in beliefs about recovery and empowerment, and greater willingness to seek treatment. TIMB is a promising stigma-reduction program and there is a need for a more detailed investigation of the program's impact using more rigorous methodology.

Knee motion variability in patients with knee osteoarthritis: The effect of self-reported instability.

BACKGROUND: Knee osteoarthritis has been previously associated with a stereotypical knee-stiffening gait pattern and reduced knee joint motion variability due to increased antagonist muscle co-contractions and smaller utilized arc of motion during gait. However, episodic self-reported instability may be a sign of excessive motion variability for a large subgroup of patients with knee osteoarthritis. The objective of this work was to evaluate the differences in knee joint motion variability during gait in patients with knee osteoarthritis with and without self-reported instability compared to a control group of older adults with asymptomatic knees. METHODS: Forty-three subjects, 8 with knee osteoarthritis but no reports of instability (stable), 11 with knee osteoarthritis and self-reported instability (unstable), and 24 without knee osteoarthritis or instability (control) underwent Dynamic Stereo X-ray analysis during a decline gait task on a treadmill. Knee motion variability was assessed using parametric phase plots during the loading response phase of decline gait. FINDINGS: The stable group demonstrated decreased sagittal-plane motion variability compared to the control group (p=0.04), while the unstable group demonstrated increased sagittal-plane motion variability compared to the control (p=0.003) and stable groups (p<0.001). The unstable group also demonstrated increased anterior-posterior joint contact point motion variability for the medial tibiofemoral compartment compared to the control (p=0.03) and stable groups (p=0.03). INTERPRETATION: The finding of decreased knee motion variability in patients with knee osteoarthritis without self-reported instability supports previous research. However, presence of self-reported instability is associated with increased knee motion variability in patients with knee osteoarthritis and warrants further investigation.

Platform for induction and maintenance of transgene-free hiPSCs resembling ground state pluripotent stem cells.

Cell banking, disease modeling, and cell therapy applications have placed increasing demands on hiPSC technology. Specifically, the high-throughput derivation of footprint-free hiPSCs and their expansion in systems that allow scaled production remains technically challenging. Here, we describe a platform for the rapid, parallel generation, selection, and expansion of hiPSCs using small molecule pathway inhibitors in stage-specific media compositions. The platform supported efficient and expedited episomal reprogramming using just OCT4/SOX2/SV40LT combination (0.5%-4.0%, between days 12 and 16) in a completely feeder-free environment. The resulting hiPSCs are transgene-free, readily cultured, and expanded as single cells while maintaining a homogeneous and genomically stable pluripotent population. hiPSCs generated or maintained in the media compositions described exhibit properties associated with the ground state of pluripotency. The simplicity and robustness of the system allow for the high-throughput generation and rapid expansion of a uniform hiPSC product that is applicable to industrial and clinical-grade use.

Myogenic differentiation of muscular dystrophy-specific induced pluripotent stem cells for use in drug discovery.

Human induced pluripotent stem cells (iPSCs) represent a scalable source of potentially any cell type for disease modeling and therapeutic screening. We have a particular interest in modeling skeletal muscle from various genetic backgrounds; however, efficient and reproducible methods for the myogenic differentiation of iPSCs have not previously been demonstrated. Ectopic myogenic differentiation 1 (MyoD) expression has been shown to induce myogenesis in primary cell types, but the same effect has been unexpectedly challenging to reproduce in human iPSCs. In this study, we report that optimization of culture conditions enabled direct MyoD-mediated differentiation of iPSCs into myoblasts without the need for an intermediate step or cell sorting. MyoD induction mediated efficient cell fusion of mature myocytes yielding multinucleated myosin heavy chain-positive myotubes. We applied the same approach to dystrophic iPSCs, generating 16 iPSC lines from fibroblasts of four patients with Duchenne and Becker muscular dystrophies. As seen with iPSCs from healthy donors, within 36 hours from MyoD induction there was a clear commitment toward the myogenic identity by the majority of iPSCs in culture (50%-70%). The patient iPSC-derived myotubes successfully adopted the skeletal muscle program, as determined by global gene expression profiling, and were functionally responsive to treatment with hypertrophic proteins insulin-like growth factor 1 (IGF-1) and wingless-type MMTV integration site family, member 7A (Wnt7a), which are being investigated as potential treatments for muscular dystrophy in clinical and preclinical studies, respectively. Our results demonstrate that iPSCs have no intrinsic barriers preventing MyoD from inducing efficient and rapid myogenesis and thus providing a scalable source of normal and dystrophic myoblasts for use in disease modeling and drug discovery.

Analysis of SOX2-expressing cell populations derived from human pluripotent stem cells.

SOX2 is involved in several cell and developmental processes, including maintenance of embryonic stem cells, differentiation of neural progenitor cells, and patterning of gut endoderm. To study its role in a human system, we generated a human embryonic stem cell (hESC) line harboring a reporter gene encoding GFP in the SOX2 locus. This SOX2 reporter line faithfully recapitulates expression of the SOX2 gene in undifferentiated human pluripotent stem cells (hPSCs), neural progenitor cells (NPCs), and anterior foregut endoderm (AFE). In undifferentiated hESCs, GFP expression corresponds to those cells with highest levels of expression of genes associated with the pluripotent state. In NPCs, expression of GFP can be employed to isolate cells expressing markers associated with NPC multipotency. In AFE, we used transcriptome-wide expression analysis to identify cell surface markers with elevated expression in this population, thereby facilitating isolation and purification of this hPSC-derived cell population.

Optimized surface markers for the prospective isolation of high-quality hiPSCs using flow cytometry selection.

hiPSC derivation and selection remains inefficient; with selection of high quality clones dependent on extensive characterization which is not amenable to high-throughput (HTP) approaches. We recently described the use of a cocktail of small molecules to enhance hiPSC survival and stability in single cell culture and the use of flow cytometry cell sorting in the HTP-derivation of hiPSCs. Here we report an enhanced protocol for the isolation of bona fide hiPSCs in FACS-based selection using an optimized combination of cell surface markers including CD30. Depletion of CD30(+) cells from reprogramming cultures almost completely abolished the NANOG and OCT4 positive sub-population, suggesting it is a pivotal marker of pluripotent cells. Combining CD30 to SSEA4 and TRA-1-81 in FACS greatly enhanced specificity and efficiency of hiPSC selection and derivation. The current method allows for the efficient and automated, prospective isolation of high-quality hiPSC from the reprogramming cell milieu.

Genetic variation of wheat streak mosaic virus in the United States Pacific Northwest.

Wheat streak mosaic virus (WSMV), the cause of wheat streak mosaic, is a widespread and damaging pathogen of wheat. WSMV is not a chronic problem of annual wheat in the United States Pacific Northwest but could negatively affect the establishment of perennial wheat, which is being developed as an alternative to annual wheat to prevent soil erosion. Fifty local isolates of WSMV were collected from 2008 to 2010 near Lewiston, ID, Pullman, WA, and the United States Department of Agriculture Central Ferry Research Station, near Pomeroy, WA to determine the amount of genetic variation present in the region. The coat protein gene from each isolate was sequenced and the data subjected to four different methods of phylogenetic analyses. Two well-supported clades of WSMV were identified. Isolates in clade I share sequence similarity with isolates from Central Europe; this is the first report of isolates from Central Europe being reported in the United States. Isolates in clade II are similar to isolates originating from Australia, Argentina, and the American Pacific Northwest. Nine isolates showed evidence of recombination and the same two well-supported clades were observed when recombinant isolates were omitted from the analysis. More polymorphic sites, parsimony informative sites, and increased diversity were observed in clade II than clade I, suggesting more recent establishment of the virus in the latter. The observed diversity within both clades could make breeding for durable disease resistance in perennial wheat difficult if there is a differential response of WSMV resistance genes to isolates from different clades.

Nutrition evidence-based guidelines for treating hyperlipidemia in children with type 1 diabetes: a case presentation.

PURPOSE: The purpose of this case presentation is to review the current nutrition evidence-based guidelines and treatment goals for hyperlipidemia in children with type 1 diabetes. The American Heart Association (AHA) places children with type 1 diabetes in the highest tier for cardiovascular risk. METHODS: Early screening for hyperlipidemia in children with diabetes is recommended to identify those children at risk. If the fasting low-density lipoprotein cholesterol (LDL-C) level is > or = 100 mg/dL (2.6 mmol/L), medical nutrition therapy is recommended as the first line of treatment to reach the desired goal (LDL-C <100 mg/dL). Medical nutrition therapy includes the following: decreasing saturated fat (<7% total calories), avoiding trans fatty acids, decreasing total cholesterol to <200 mg daily, increasing soluble fiber, and adding phytosterols daily. RESULTS: The patient discussed in this case presentation achieved a desired LDL-C level <100 mg/dL (2.6 mmol/L) by following the recommended heart-healthy guidelines. Statin therapy was not considered unless the LDL-C goal, <130 mg/dL (3.38 mmol/L), was not achieved by diet alone. CONCLUSIONS: In this case study, evidence-based nutrition guidelines have been evaluated and reviewed to demonstrate heart-healthy eating for children with hyperlipidemia and type 1 diabetes. It is known that approximately 40% to 50% of children with elevated lipids will continue to have abnormal lipids into adolescence and early adulthood. Therefore, early screening is recommended by the AHA to track lipid changes during childhood and adolescence and to begin treating abnormal LDL-C levels to prevent the development of atherosclerosis.

Cloning and expression of human mitogen-activated protein kinase kinase 7gamma1.

Mitogen-activated protein kinase kinase 7 (MKK7) is a direct activator of the mitogen-activated protein kinase family member c-Jun N-terminal kinase (JNK). MKK7 activates JNK via phosphorylation of a threonine and tyrosine residue in a Thr-Pro-Tyr motif within kinase subdomain VIII. To date at least six different isoforms of murine MKK7 have been identified. However, only three isoforms of human MKK7 have been reported. We report here the cloning of hMKK7gamma1, the human homolog of murine MKK7gamma1. Expression of hMKK7gamma1 mRNA was assessed and transcripts were present in low levels in placenta, fetal liver, and skeletal muscle. PCR results indicate that hMKK7gamma1 is expressed in various normal tissues, tumors, and in synoviocytes from rheumatoid and osteoarthritis patients. Recombinant hMKK7gamma1 can be phosphorylated and activated by MEKK1. Further studies will provide insight into the role for hMKK7gamma1 versus other MKK7 isoforms.