Corneal Endothelial Cell Cultures from Organotypic Preservation of Older Donor Corneas Are Suitable for Advanced Cell Therapy.

INTRODUCTION: The purpose of this work was to evaluate the in vitro growth capacity and functionality of human corneal endothelial cells (hCEC) expanded from corneas of elderly (>60 years) donors that were preserved using an organotypic culture method (>15 days, 31°C) and did not meet the clinical criteria for keratoplasty. METHODS: Cell cultures were obtained from prior descemetorhexis (≥10 mm) and a controlled incubation with collagenase type I followed by recombinant trypsin. Cells were seeded on coated plates (fibronectin-albumin-collagen I) and cultures were expanded using the dual supplemented medium approach (maintenance medium and growth medium), in the presence of a 10 µm Rho-associated protein kinase inhibitor (Y-27632). Cell passages were obtained at culture confluency (∼2 weeks). A quantitative colorimetric WST-1 cell growth assay was performed at different time points of the culture. Morphometric analysis (area assessment and circularity), immunocytochemistry (ZO-1, Na+/K+-ATPase α, Ki67), and transendothelial electrical resistance (TEER) were performed on confluent monolayers. RESULTS: There was no difference between the cell growth profiles of hCEC cultures obtained from corneas older than 60 years, whether preserved cold or cultivated organotypic corneas. Primary cultures were able to maintain a certain cell circularity index (around 0.8) and morphology (hexagonal) similar to corneal endothelial mosaic. The ZO-1 and Na+/K+-ATPase pump markers were highly positive in confluent cell monolayers at 21 days after isolation (passage 0; P0), but significantly decreased in confluent monolayers after the first passage (P1). A weak expression of Ki67 was observed in both P0 and P1 monolayers. The P0 monolayers showed a progressive increase in TEER values between days 6 and 11 and remained stable until day 18 of culture, indicating a state of controlled permeability in monolayers. The P1 monolayers also showed some functional ability but with decreased TEER values compared to monolayers at P0. CONCLUSIONS: Our results indicate that it is possible to obtain functional hCEC cultures in eye banks, using simplified and standardized protocols, from older donor corneas (>60 years of age), previously preserved under organotypic culture conditions. This tissue is more readily available in our setting, due to the profile of the donor population or due to the low endothelial count (<2,000 cells/mm2) of the donated cornea.

A Framework for Human Corneal Endothelial Cell Culture and Preliminary Wound Model Experiments with a New Cell Tracking Approach.

Cell injection therapy is emerging as an alternative to treat corneal endothelial dysfunction (CED) and to avoid corneal scarring due to bullous keratopathy. However, establishing a standardized culture procedure that provides appropriate cell yield while retaining functional features remains a challenge. Here, we describe a detailed framework obtained from in vitro culture of human corneal endothelial cells (HCECs) and comparative in vivo experimental models for CED treatment with a new cell tracking approach. Two digestion methods were compared regarding HCEC morphology and adhesion. The effect of Y-27632 (ROCKi) supplementation on final cell yield was also assessed. Cell adhesion efficacy with two cell delivery systems (superparamagnetic embedding and cell suspension) was evaluated in an ex vivo human cornea model and in an in vivo rabbit CED model. The injection of supplemented culture medium or balanced salt solution (BSS) was used for the positive and negative controls, respectively. HCEC isolation with collagenase resulted in better morphology and adhesion of cultured HCEC when compared to EDTA. Y-27632 supplementation resulted in a 2.6-fold increase in final cell yield compared to the control. Ex vivo and in vivo adhesion with both cell delivery systems was confirmed by cell tracker fluorescence detection. Corneal edema and opacity improved in both animal groups treated with cultured HCEC. The corneas in the control groups remained opaque. Both HCEC delivery systems seemed comparable as treatments for CED and for the prevention of corneal scarring.

Fracture Risk After Bariatric Surgery: A Systematic Literature Review and Meta-Analysis.

OBJECTIVE: Bariatric surgery has undeniable benefits for cardiovascular and metabolic health, but it can result in negative effects on mineral and bone metabolism. This study aimed to review the current data on fractures after the main types of bariatric surgery. METHODS: Therefore, a systematic review was performed using the electronic literature available in the databases LILACS, CENTRAL, Web of Science, Embase, and PubMed/MEDLINE, with results until January 2021. Keywords were descriptors for bariatric surgery or bariatric medicine or bariatrics or gastroplasty and bone or bones or bone fractures, referenced in all words of the text in the study. RESULTS: The meta-analysis of observational studies and interventional trials showed that the risk of any type of fracture was higher in the surgical group than in the nonsurgical group (relative risk [RR], 1.20 [95% confidence interval {CI}, 1.15-1.26; P < .00001; Phetero < 0.000001; I2 = 94%], and RR, 1.16 [95% CI, 1.00-1.33; P = .04; Phetero = 0.27; I2 = 23%], respectively). It is worth noting that the included interventional studies have a low rating on the risk of bias assessment scales. Compared with those who underwent restrictive procedures, subjects had an increased fracture risk after malabsorptive procedures (RR, 0.49 [95% CI, 0.40-0.61; P < .00001); Phetero = 0.96; I2 = 0%). CONCLUSION: We found that bariatric procedures are associated with an increased risk of fractures, especially the malabsorptive techniques.

Agreement Between American and European Thyroid Imaging, Reporting, and Data System (TIRADS) in the Diagnosis of 473 Thyroid Nodules From a Single Center in Brazil.

OBJECTIVE: To compare 2 ultrasound-based risk stratification systems in malignancy risk assessment of thyroid nodules and the clinical applicability of these guidelines in Brazil. METHODS: We retrospectively reviewed the ultrasound findings of 314 patients (473 thyroid nodules) who underwent fine-needle aspiration (FNA) biopsy and/or surgery between February 2018 and March 2019. All nodules were classified using 2 systems: the Thyroid Imaging, Reporting, and Data System (TIRADS) of the American College of Radiology (ACR-TIRADS) and the TIRADS of the European Thyroid Association (EU-TIRADS). Both risk stratification systems were analyzed. We identified the diagnostic predictive values that yielded optimal sensitivity, specificity, positive predictive value, negative predictive value, and accuracy. RESULTS: Of the 473 nodules, all underwent FNA, and histopathology was performed for 332 nodules. The agreement between the ACR-TIRADS and EU-TIRADS results and that between cytology and histopathology findings was 92.6% (kappa = 0.84) and 86.7% (kappa = 0.73), respectively. The area under the curve for the ACR-TIRADS and EU-TIRADS was 0.871 and 0.828, respectively (P < .001). The EU-TIRADS had the best sensitivity and negative predictive value, whereas the ACR-TIRADS had the best specificity, positive predictive value, and accuracy. Of the 473 nodules studied, only 158 (33.4%) followed the FNA size criteria suggested by the ACR-TIRADS. CONCLUSION: ACR-TIRADS and EU-TIRADS had good diagnostic performances. However, most aspirated nodules did not follow the TIRADS indication; thus, the overuse of FNA as a diagnostic tool was observed.

Vitamin D deficiency and visceral adipose tissue in early pregnant women.

BACKGROUND: We aimed to assess the correlation between vitamin D serum level and visceral fat tissue during early pregnancy. METHODS: This cross-sectional study was performed in Pernambuco, Brazil. 190 low risk pregnant women (8-16 gestational weeks) were eligible. Visceral adipose tissue was measured by ultrasonography following the technique described by Armellini. The 25(OH) D in serum was determined through chemiluminescence. The Spearman correlation test was applied to evaluate the correlation between vitamin D serum level and VAT, considering p < 0.05 to be significant. RESULTS: Vitamin D insufficiency was present in 129 (67.8 %) of subjects. Pregnant women with or without vitamin D deficiency did not differ in age, gestational age, nutritional status and visceral adipose tissue. No correlation between visceral adipose tissue and 25(OH) D was observed: - 0.057 (p = 0.435). CONCLUSIONS: Maternal visceral adipose tissue and vitamin D serum level are not correlated during pregnancy.

High Rate of Occult Urolithiasis in Normocalcemic Primary Hyperparathyroidism.

INTRODUCTION: Normocalcemic primary hyperparathyroidism (NPHPT) is characterized by elevations in serum parathyroid hormone levels in the presence of normal serum calcium concentrations after exclusion of secondary hyperparathyroidism. We have previously demonstrated no differences in the prevalence of clinically active urolithiasis between NPHPT and hypercalcemic asymptomatic PHPT, and that it is significantly higher in postmenopausal osteoporotic women with NPHPT in comparison to women with normal serum PTH and calcium concentrations. Few studies have addressed the occurrence of silent or occult kidney stones in asymptomatic hypercalcemic PHPT, but no data are available for NPHPT. OBJECTIVE: To determine the presence of occult urolithiasis in NPHPT patients using routine abdominal ultrasonography. METHODS AND RESULTS: We studied 35 patients with NPHPT (mean age 63.2 ± 10.7 years, 96% women; serum PTH 116.5 ± 39.2 pg/mL, 25OHD 38.5 ± 6.82 ng/mL, total calcium 9.1 ± 0.56 mg/dL; albumin 4.02 ± 0.37 g/dL; BUN 34.35 ±10.23 mg/dL; p = 3.51 ± 0.60 mg/dL; estimated glomerular filtration rate 88.44 ± 32.45 mL/min/1.73 m2, and 24-h urinary calcium excretion 140.6 ± 94.3 mg/24 h). The criteria for the diagnosis of NPHPT were as follows: serum PTH above the reference range (11-65 pg/mL), normal albumin-corrected serum calcium concentrations, normal 24-h urinary calcium excretion, serum 25OHD above 30 ng/mL, estimated GFR (MDRD) above 60 mL/min/1.73 m2 (with the exclusion of medications such as thiazide diuretics, lithium, bisphosphonates, and denosumab), a history of clinical symptoms of urolithiasis, and a family history of kidney stones. Thirty-five patients were evaluated and 25 of them met the inclusion criteria. Five patients presented nephrolithiasis corresponding to 20% of the study population. There were no statistically significant differences in any of the clinical or laboratory variables studied between patients with or without urolithiasis, although mean serum PTH levels were higher in patients with stones (180.06 ± 126.48 vs. 100.72 ± 25.28 pg/mL, p = 0.1). The size of the stones ranged from 0.6 to 0.9 cm and all of the stones were located in the renal pelvis. CONCLUSION: We found a high prevalence of occult kidney stones in NPHPT patients, similar to what is observed in clinically manifested urolithiasis, in hypercalcemic PHPT.

Multiple severe vertebral fractures during the 3-month period following a missed dose of denosumab in a postmenopausal woman with osteoporosis previously treated with alendronate
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Denosumab is a monoclonal antibody that decreases bone resorption and increases bone mass and strength in trabecular and cortical bone leading to a reduction in fracture in women and men. Its effects are reversed after discontinuation. Recently, there has been concern about the discontinuation of denosumab and the increased risk of multiple vertebral fractures that would be associated with rapid bone loss due to high bone turnover and consequent increase in markers of bone remodeling, far above the baseline values. Some case reports have demonstrated the occurrence of more than two fractures within a period of 8 - 16 months after the drug's discontinuation. A 56-year-old female presented with multiple vertebral fractures in a short period of time after denosumab withdrawal. She took alendronate for 12 years since the occurrence of a rib fracture (from the age of 30 to 42 years). After a long period with no medication (12 years), she was placed on denosumab. On the day she was due to take the fourth injection, she had sudden back pain, and magnetic resonance imaging (MRI) showed a grade II fracture at T9. She had simply forgot to take the scheduled dose of denosumab. Three months later, she presented with several episodes of severe back pain and loss of height and was diagnosed with multiple new severe vertebral fractures. This case illustrates an uncommon and severe bone disease following the inadvertent discontinuation of denosumab treatment.
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Bone Density in Patients With Berardinelli-Seip Congenital Lipodystrophy Is Higher in Trabecular Sites and in Type 2 Patients.

Berardinelli-Seip congenital lipodystrophy (BSCL) is a rare autosomal recessive syndrome characterized by a difficulty storing lipid in adipocytes, low body fat, hypoleptinemia, and hyperinsulinemia. We report here laboratory, bone mineral density (BMD), and bone mineral content findings of 21 patients (24.1 ± 8.4 yr old, 14 females, 18 diabetics, 5.3% total body fat) with BSCL. The mean leptin was very low (0.91 ± 0.42 ng/mL), and the mean values of the Z-scores for all studied sites were positive, except for the 33% radius (Z-score -0.5 standard deviation [SD]). Twelve patients (57.1%) had a BMD Z-score higher than +2.5 SD in at least 1 site. There was no significant difference in the Z-scores between males and females. None of type 1 (AGPAT2) patients had Z-scores higher than +2.5 SD, and these patients had a smaller Z-score of BMD total body (0.26 SD vs 1.90 SD, p = 0.022) and of bone mineral content (1.59 SD vs 3.3 SD, p = 0.032) than type 2 (seipin) patients. Insulin, as well as HOMAIR (homeostasis model assessment), correlated positively with the BMD of all sites, except for the 33% radius. Z-Scores on this site (33% radius) were the smallest of all. More than half of our patients with BSCL have BMD Z-scores higher than +2.5 SD on at least 1 site, and this increase is more pronounced in the trabecular sites and in type 2 patients.

Hyperparathyroidism and Bone Health.

Bone pain, proximal muscle weakness, skeletal deformities, and pathological fractures are features of osteitis fibrosa cystica which occur in severe primary hyperparathyroidism (PHPT). In this condition, bone mineral density is usually extremely low, but may be reversible after parathyroidectomy. On X-ray, bone abnormalities are described as having a salt-and-pepper appearance in the skull, with bone erosions and resorption of the phalanges, brown tumors and cysts, as well as diffuse demineralization, along with pathological fractures, particularly in the long bones of the extremities. A marked elevation of the serum calcium and PTH concentrations is seen, and renal involvement is manifested by nephrolithiasis and nephrocalcinosis. In asymptomatic PHPT, the absence of clinically significant bone involvement has led to much more data on bone mineral density becoming available by dual X-ray absorptiometry (DXA) and also on new technologies such as trabecular bone score (TBS), which is a gray-level textural analysis of DXA images that provides an indirect index of trabecular microarchitecture. In addition, high-resolution peripheral computed tomography (HRpQCT), which has a low radiation exposure, provides further understanding of the microstructural skeletal features at both trabecular and cortical sites.

Prevalence of morphometric vertebral fractures in old men and the agreement between different methods in the city of Recife, Brazil.

Osteoporosis is relatively common in men and has a great impact on quality of life. Despite the importance of the subject, there are few studies regarding the prevalence of morphometric vertebral fractures in men and the associated risk factors. To determine the prevalence of morphometric vertebral fractures in elderly men by three different methods and the agreement between them, 234 asymptomatic men aged >60 years (mean age 69.4 ± 6.5 years) were evaluated using lateral thoracolumbar radiograph that were analyzed by two experienced radiologists according to semiquantitative (SQ) Genant and algorithm-based qualitative (ABQ) Jiang methods. A third senior radiologist adjudicated Genant's method. The highest prevalence of fractures in ABQ Jiang and SQ Genant methods were 37.6 and 36.8 %, respectively (both examiner 2). The lowest prevalence rates were 26.5 % in ABQ Jiang method and 5.6 % in SQ Genant (both examiner 1). The prevalence found by the Genant adjudicated was 31.6 %. The agreement between the examiners were 69.2 % in ABQ Jiang method (κ 0.30; 95 % CI 0.17-0.43) and 65.5 % in SQ Genant (κ 0.09; 95 % CI 0.01-0.17). We evaluated skin phototype, waist circumference, hypertension, body mass index (BMI), history of fracture, calcium intake, serum 25 OHD and sun index. After multivariate regression analysis, we found that lower BMI (prevalence ratio = 1.41; p = 0.024; 95 % CI 1.05-2.03) and sun index (prevalence ratio = 1.45; p = 0.049; 95 % CI 1.01-1.95) were independently associated with morphometric vertebral fractures.

Vitamin D Deficiency in Patients Hospitalized for Heart Failure Living in the Tropics.

Background and Objectives: Vitamin D, as a steroid hormone, has multiple effects on human body and its deficiency has been associated with an increased risk of heart failure (HF) and unfavorable outcomes. The present study investigated the prevalence of vitamin D deficiency (VDD) and its relationship with cardiometabolic parameters in patients hospitalized for HF living in the city of Recife (latitude 8° South). Methods: Analytical cross-sectional study, with men and women aged 40-64 years. The HF group was recruited during hospitalization due to decompensation. A matched control group was recruited from the general endocrine clinics. Vitamin D status was assessed by measuring serum 25-hydroxyvitamin D (25OHD), considering deficiency when 25OHD <20 ng/mL (<50 nmol/L). Results: A total of 243 patients were evaluated (HF group: 161, control group: 82). Lower serum 25OHD levels were observed in the HF group (25.2±9.4 vs. 30.0±7.7ng/mL; p<0.001), as well as a higher prevalence of VDD (27.3% vs. 9.8%; prevalence ratio, 2.80; 95% confidence interval, 1.38-5.67; p=0.002). In patients with HF, VDD was associated with diabetes mellitus (65.9% vs. 41.0%; p=0.005) and female sex (65.9% vs. 44.4%; p=0.015). In the subgroup with VDD, higher values of hemoglobin A1c (7.9% [6.0-8.9] vs. 6.2% [5.7-7.9]; p=0.006) and dyslipidemia were also observed. Conclusions: We found higher rates of VDD in patients hospitalized for HF and this was associated with deleterious laboratory metabolic parameters.

Comparison of intraocular lens tilt after capsular sutured scleral fixation with capsular segments versus uneventful cataract surgery.

PURPOSE: To evaluate and compare intraocular lens (IOL) tilt between uneventful phacoemulsification with in-the-bag IOL implantation and sutured scleral fixation (SSF) of the lens bag with a capsular tension segment (type 6 D / Morcher) using a Sheimpflug camera. SETTING: Clinical Practice, Hospital. Barcelona and A Coruña, Spain. DESIGN: Retrospective, comparative multicenter study. METHODS: IOL tilt was compared between patients who underwent sutured scleral fixation with a capsular tension segment in a single eye (SSF group, n = 15) with patients who underwent uneventful IOL implantation (control group, n = 12) that were matched by biometric measurements. Post-operative refractive accuracy of biometric formulas by means of mean absolute error (MAE) was also reported. All patients underwent a general ophthalmic evaluation, anterior segment photography, and postoperative optical biometry (Zeiss IOLMaster® 500). In addition, IOL tilt was measured with a Scheimpflug camera (Pentacam R, Oculus Optikgerate Gmbh). RESULTS: Mean vertical tilt was similar in both groups (2.20+/-2.47° SSF vs 1.97 +/- 1.79° control; p = 0.836) but mean horizontal tilt tended to higher values in the SSF series (2.09 +/- 2.74° vs 0.94 +/- 1.17°; p = 0.139). Considering post-operative refractive error in diopters by MAE calculations, there was an underestimation of IOL power in the SSF group which was only statistically significant for Barrett Universal II (1.07 vs 0.32; p = 0.028) and Hill-RBF (0.95 vs 0.26; p = 0.024) formulas, but not for SRK/T (0.99 vs 0.42; p = 0.285) and Kane (0.96 vs 0.33; p = 0.083). CONCLUSION: Sutured scleral fixation of capsular tension segments in the presence of zonular instability does not seem to induce clinically significant IOL tilt compared to uneventful cataract extraction cases.

Prevalence of Morphometric Vertebral Fractures After Bariatric Surgery and Its Relationship with Bone Mineral Density and Bone Markers.

BACKGROUND: Bariatric surgery (BS) can lead to bone loss and an increased fracture risk. METHODS: To determine the morphometric vertebral fracture (MVF) prevalence, and its relationship with bone mineral density (BMD), and biomarker's turnover after Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy (SG), we analyzed post-surgery X-rays of the spine in 80 patients (88% female, 51% RYGB, age 41.2 [6.8] years) from 117 participants' retrospective cohort (1-2 years, >2 and <5 years, and >5 years). We still analyzed body composition and BMD by dual-energy X-ray absorptiometry and bone parameters. RESULTS: MVF prevalence was 17.5% (14/80), with no statistical difference between groups (p = 0.210). RYGB group had a higher prevalence of secondary hyperparathyroidism (SHPT) (PTH ≥ 65 pg/ml; 18.4% vs 7.8%, respectively, p = 0.04), PTH (61.3 vs 49.5 pg/ml, p = 0.001), CTX (0.766 [0.29] ng/ml vs 0.453 [0.30] ng/ml, p = 0.037), and AP (101.3 [62.4] U/L vs 123.9 [60.9] U/L, p = 0.027) than the SG group. Up to 5 years postoperatively, RYGB had a lower total (1.200 [0.087] vs 1.236 [0.100] g/cm2, p = 0.02), femoral neck (1.034 [0.110] vs 1.267 [0.105], p = 0.005), and total femur BMD (1.256 [0.155] vs 1.323 [0.167], p = 0.002) than SG group. We found no statistically significant difference between the MFV (+) and MVF (-) groups regarding age, sex, BMI, surgery time, BMD, or bone and metabolic parameters, including leptin. CONCLUSION: We found a high prevalence of MVF after BS with no differences between RYGB and SG.

Metabolic syndrome and central fat distribution are related to lower serum osteocalcin concentrations.

BACKGROUND: The aim of this study was to evaluate the association of serum osteocalcin with the metabolic syndrome (MS) in men and premenopausal women. METHODS: 14 middle-aged men and 44 premenopausal women were evaluated. MS was defined according to the International Diabetes Federation criteria. Anthropometric data were collected and serum osteocalcin, serum C-telopeptide (CTX), fasting plasma glucose (FPG) and lipid profile measured. RESULTS: The mean age was 41.07 ± 8.4 years and did not differ between patients with and without MS. Mean osteocalcin was significantly lower in patients with MS (11.18 ± 4.62 vs. 15.09 ± 5.05, p = 0.003) and decreased significantly with the rise in the number of criteria for diagnosis of MS. There were no significant differences in serum CTX between the two groups. Serum osteocalcin was lower in patients with body mass index (BMI) ≥25 (p = 0.038) and FPG ≥100 mg/dl (p = 0.024), and in hypertensive (p = 0.013) and diabetic patients (p = 0.036), and was inversely associated with BMI (p = 0.024), waist circumference (WC) (p = 0.024), FPG (p = 0.007) and systolic blood pressure (SBP) (p = 0.037). CONCLUSION: This study showed that lower serum osteocalcin is associated with the presence of MS and that osteocalcin is inversely associated with BMI, WC, FPG and SBP, suggesting that osteocalcin plays a part in the development of MS.

Hypopituitarism due to a Large Osteoclastoma Arising from the Sphenoid Bone Invading the Pituitary Fossa in a Patient with Parathyroid Carcinoma.

Background: Parathyroid carcinoma accounts for <1% of cases of primary hyperparathyroidism (PHPT). This rare condition may present with severe hypercalcemia and bone complications such as osteoclastomas and pathologic fractures. Here, we present a rare condition of panhypopituitarism resulting from an osteoclastoma in the sphenoid bone that invaded the pituitary fossa due to parathyroid carcinoma. Case Report. A 47-year-old woman previously diagnosed with PHPT underwent a parathyroidectomy 6 years earlier, with histological examination indicating a parathyroid adenoma. After surgery, she continued to exhibit high serum parathyroid hormone (PTH) and calcium levels, with the development of bone pain and spontaneous fractures. Imaging exams showed a large osteoclastoma of the sphenoid bone, invading the pituitary fossa, causing hypopituitarism. A new parathyroidectomy was performed, with histological confirmation of parathyroid carcinoma and regression of the osteoclastoma. Conclusion: This case illustrates an unusual presentation of parathyroid carcinoma, in which an osteoclastoma of the sphenoid bone caused hypopituitarism.

Bone Mineral Density, Trabecular Bone Score and Fractures in Patients Hospitalized for Heart Failure.

BACKGROUND: This study aimed to evaluate the bone mineral density (BMD), trabecular bone score (TBS), and fracture history of middle-aged patients hospitalized for heart failure (HF), as well as analyze the association of these factors with cardiometabolic parameters and muscle strength. METHODS: A cross-sectional study with patients aged 40 to 64 years hospitalized for HF was performed. Dual energy X-ray absorptiometry was performed to obtain BMD and TBS. Fracture history, handgrip strength (HGS), and clinical and laboratory cardiometabolic parameters of the patients were evaluated. RESULTS: Altogether, 109 patients were evaluated (female 50.5%). Medians and interquartile ranges for age and length of hospital stay were 58.0 (53.0-61.0) years and 20.0 (11.0-32.0) days, respectively. Osteoporosis was observed in 15.6% of the patients, low TBS was observed in 22.8%, and 6 patients had a history of fragile fracture. No differences between the sexes regarding BMD (p=0.335) or TBS (p=0.736) classifications were observed. No association was observed between low BMD and HF classification (p>0.05) regarding the ejection fraction, ischemic etiology, or New York Heart Association Functional Classification. However, there was a significant association between high serum parathyroid hormone (PTH) and the presence of osteoporosis (62.5 [37.2-119.0] pg/mL vs. 34.2 [25.0-54.1] pg/mL; p=0.016). There was a negative correlation between serum PTH and TBS (r=-0.329, p=0.038) and a higher frequency of reduced HGS in patients with low TBS (92.3% vs. 50.0%; p=0.009). CONCLUSIONS: We found relevant frequencies of osteoporosis and bone microarchitecture degradation in middle-aged patients with HF, which were related to high serum PTH concentrations.

Effects of Rho-Associated Kinase (Rock) Inhibitors (Alternative to Y-27632) on Primary Human Corneal Endothelial Cells.

(1) Rho-associated coiled-coil protein kinase (ROCK) signaling cascade impacts a wide array of cellular events. For cellular therapeutics, scalable expansion of primary human corneal endothelial cells (CECs) is crucial, and the inhibition of ROCK signaling using a well characterized ROCK inhibitor (ROCKi) Y-27632 had been shown to enhance overall endothelial cell yield. (2) In this study, we compared several classes of ROCK inhibitors to both ROCK-I and ROCK-II, using in silico binding simulation. We then evaluated nine ROCK inhibitors for their effects on primary CECs, before narrowing it down to the two most efficacious compounds-AR-13324 (Netarsudil) and its active metabolite, AR-13503-and assessed their impact on cellular proliferation in vitro. Finally, we evaluated the use of AR-13324 on the regenerative capacity of donor cornea with an ex vivo corneal wound closure model. Donor-matched control groups supplemented with Y-27632 were used for comparative analyses. (3) Our in silico simulation revealed that most of the compounds had stronger binding strength than Y-27632. Most of the nine ROCK inhibitors assessed worked within the concentrations of between 100 nM to 30 µM, with comparable adherence to that of Y-27632. Of note, both AR-13324 and AR-13503 showed better cellular adherence when compared to Y-27632. Similarly, the proliferation rates of CECs exposed to AR-13324 were comparable to those of Y-27632. Interestingly, CECs expanded in a medium supplemented with AR-13503 were significantly more proliferative in (i) untreated vs. AR-13503 (1 µM; * p < 0.05); (ii) untreated vs. AR-13503 (10 µM; *** p < 0.001); (iii) Y-27632 vs. AR-13503 (10 µM; ** p < 0.005); (iv) AR-13324 (1 µM) vs. AR-13503 (10 µM; ** p < 0.005); and (v) AR-13324 (0.1 µM) vs. AR-13503 (10 µM; * p < 0.05). Lastly, an ex vivo corneal wound healing study showed a comparable wound healing rate for the final healed area in corneas exposed to Y-27632 or AR-13324. (4) In conclusion, we were able to demonstrate that various classes of ROCKi compounds other than Y-27632 were able to exert positive effects on primary CECs, and systematic donor-match controlled comparisons revealed that the FDA-approved ROCK inhibitor, AR-13324, is a potential candidate for cellular therapeutics or as an adjunct drug in regenerative treatment for corneal endothelial diseases in humans.

Effects of Preoperative Sarcopenia-Related Parameters on Cardiac Autonomic Function in Women with Obesity Following Bariatric Surgery: A One-Year Prospective Study.

OBJECTIVES: Investigate changes in blood pressure (BP) and heart rate variability (HRV) in women with and without sarcopenia-related parameters who underwent bariatric surgery (BS) during a one-year follow-up. SUBJECTS AND METHODS: Women were separated into obesity (OB, n = 20) and women with obesity displaying sarcopenia-related parameters (SOP, n = 14) and evaluated before BS and 3, 6, and 12 months after BS. SOP was defined as low handgrip strength (HS) and/or low appendicular skeletal mass adjusted for weight (ASM/wt × 100, %) in the lowest quartile of the sample. ASM/wt × 100, % and HS were significantly lower in SOP than OB over a one-year follow-up of BS (p < 0.05). RESULTS: There was a reduction in diastolic BP, heart rate (HR), SDHR, LF, and the LF/HF ratio (p < 0.05) and an increase in the HF band in both groups during the follow-up period (p < 0.05). SOP women had reduced root mean square differences of successive RR intervals (RMSSD) and HF band and an increased LF band and SD2/SD1 ratio compared to the OB group during the one-year follow-up (p < 0.05). ASM/wt × 100, % was negatively associated with the LF band (r = -0.24, p = 0.00) and positively associated with the HF band (r = 0.22, p = 0.01). Conversely, HS had no association with LF (r = -0.14, p = 0.09) and HF (r = 0.11, p = 0.19). ASM/wt × 100, % and HS were negatively associated with the LF/HF ratio (p < 0.05). CONCLUSIONS: Women who underwent BS had an improved HRV over a one-year follow-up. However, the improvement in HRV variables was less pronounced in women with low muscle mass and/or HS during the follow-up period.

Vitamin D Concentration Changes after Bariatric Surgery.

Introduction: Bariatric surgery causes physiological and anatomical changes in the gastrointestinal tract that interfere with intestinal absorption and, consequently, with the nutritional status, especially about vitamin D. The aim of the study was to evaluate the vitamin D levels and body composition of these patients in the Roux-en-Y gastric bypass (RYGB) and vertical sleeve gastrectomy (VSG) types of surgery. Methods: The retrospective cohort study included a population of 120 patients aged between 18 and 65 years, with class II or III obesity, undergoing bariatric surgery procedures (VSG or RYGB-type). Data were collected on the degree of obesity, age, average time since surgery, and gender. The individuals underwent a complete physical examination, measuring blood pressure, weight, height, waist, and neck circumference. In addition to calculating the percentage of loss of body weight and assessing body fat, the food frequency and physical activity of these patients were evaluated. Blood was collected, and the insulin variables, hydroxyvitamin D (25OHD), were analyzed. Results: There was a significant difference between groups only for PTH, total BMD, and insulin variables. A significant intragroup difference was found in the variables' body mass index (BMI) and vitamin D for the vertical sleeve gastrectomy group and BMI for the RYGB group. Conclusion: The analysis between the groups of procedures, similarity in body composition and postsurgical vitamin D levels, with significant differences only for PTH, BMD, and insulin variables, demonstrates that both procedures are effective in reducing fat mass.

Association between vascular calcification and osteoporosis in men with type 2 diabetes.

Atherosclerotic vascular disease is common in diabetes, and some data support a link with bone loss. This study evaluates the association between osteoporosis and clinical and metabolic factors and chronic complications of diabetes. We studied 59 diabetic men aged 50-80 yr who were assessed with bone densitometry (dual-energy X-ray absorptiometry). Of them, 10.2% of the patients were found to have osteoporosis in the lumbar spine and 45.8% osteopenia, whereas in the femoral neck, 11.8% had osteoporosis and 49% had osteopenia. There was a significant association of osteoporosis in the lumbar spine L1-L4 (p=0.004) and in the femoral neck (p=0.036) with iliac artery calcification. In addition, there was no association with any other metabolic factors, clinical factors, or chronic complications of diabetes evaluated, except for an association between a previous personal history of fractures (p=0.016) and low bone mineral density in the femoral neck. In conclusion, we found a positive association between the iliac artery calcification and osteoporosis in type 2 diabetic male patients.