Dapagliflozin administration to a mouse model of type 2 diabetes induces DNA methylation and gene expression changes in pancreatic islets.

Decreased pancreatic ß-cell volume is a serious problem in patients with type 2 diabetes mellitus, and there is a need to establish appropriate treatments. Increasingly, sodium/glucose cotransporter 2 (SGLT2) inhibitors, which have a protective effect on pancreatic ß-cells, are being prescribed to treat diabetes; however, the underlying mechanism is not well understood. We previously administered SGLT2 inhibitor dapagliflozin to a mouse model of type 2 diabetes and found significant changes in gene expression in the early-treated group, which led us to hypothesize that epigenetic regulation was a possible mechanism of these changes. Therefore, we performed comprehensive DNA methylation analysis by methylated DNA immunoprecipitation using isolated pancreatic islets after dapagliflozin administration to diabetic model mice. As a result, we identified 31 genes with changes in expression due to DNA methylation changes. Upon immunostaining, cystic fibrosis transmembrane conductance regulator and cadherin 24 were found to be upregulated in islets in the dapagliflozin-treated group. These molecules may contribute to the maintenance of islet morphology and insulin secretory capacity, suggesting that SGLT2 inhibitors' protective effect on pancreatic ß-cells is accompanied by DNA methylation changes, and that the effect is long-term and not temporary. In future diabetes care, SGLT2 inhibitors may be expected to have positive therapeutic effects, including pancreatic ß-cell protection.

Leaf spring exercise: A safe quadriceps strengthening exercise after anterior cruciate ligament reconstruction.

BACKGROUND: Leg extensions should be avoided in the early stages after anterior cruciate ligament reconstruction because the force exerted by the quadriceps muscle leads to anterior tibial displacement. To allow for safe quadriceps training in the knee extension range during this period, we devised the leaf spring exercise, which involves placing subjects in the prone position with their knee slightly flexed and instructing them to perform maximum isometric quadriceps contractions while supporting the proximal region of the lower leg's anterior surface and immobilizing the femur's posterior surface to prevent lifting. The current study aimed to examine the safety of Leaf spring exercise by determining the femur-tibia relationship using ultrasound imaging. METHODS: This controlled laboratory study included patients with unilateral anterior cruciate ligament-deficient knees (8 men and 8 women; age, 24.2 ± 8.3 years) who were instructed to perform Leaf spring exercise of both lower limbs. We measured the femur-tibia-step-off, which indicates the distance between the last point of the medial and lateral condyles of the femur and posterior margin of the tibial plateau, as a parameter to evaluate anterior tibial displacement via ultrasound diagnostic device. Further, peak torque of the quadriceps muscle was calculated using force measurement device. FINDINGS: No difference in anterior tibial displacement and peak torque was observed between the uninjured and injured sides during Leaf spring exercise. INTERPRETATION: Leaf spring exercise may add some strain on the reconstructed anterior cruciate ligament; hence, it can be considered a safe quadriceps exercise in the knee extension range.

Distinct hypoglycemic effect of different formulations of a fixed ratio of basal insulin plus glucagon-like peptide-1 receptor agonist in a patient with pancreatic diabetes.

Fixed-ratio combination injection therapy (FRC) is a fixed-ratio mixture containing basal insulin and glucagon-like peptide-1 receptor agonist (GLP-1 RA) in a single injection for the treatment of patients with type 2 diabetes. The two types of FRC products contain different concentrations and mixing ratios of basal insulin and GLP-1 RA. Both products demonstrated satisfactory blood glucose control throughout the day, with less hypoglycemia and weight gain. However, few studies have examined the differences in the actions of the two formulations. Herein, we present a case of a 71-year-old man with pancreatic diabetes and significantly impaired intrinsic insulin secretion capacity, who demonstrated a marked difference in glycemic control following treatment with two different FRC formulations. Treatment with IDegLira, an FRC product, demonstrated suboptimal glucose control in the patient. However, after a change in therapy to another FRC product, IGlarLixi, his glucose control markedly improved, even with a decrease in the injection dose. This difference could have been due to lixisenatide, a short-acting GLP-1RA contained in IGlarLixi, which exerts a postprandial hypoglycemic effect irrespective of intrinsic insulin secretion capacity. In conclusion, IGlarLixi has the potential to achieve good fasting and postprandial glucose control with a once-daily injection, even in patients with type 2 diabetes who have a reduced intrinsic insulin secretion capacity. Supplementary Information: The online version contains supplementary material available at 10.1007/s13340-023-00621-5.

Chlorogenic Acid and Caffeine in Coffee Restore Insulin Signaling in Pancreatic Beta Cells.

The incidence of type 2 diabetes is reported to be lower in frequent coffee drinkers than in non-coffee drinkers. To elucidate the mechanism by which coffee prevents the onset of type 2 diabetes, we analyzed how caffeine and chlorogenic acid, which are components of coffee, alter insulin signaling in MIN6 cells, a mouse pancreatic Β cell line. The results showed that caffeine improved insulin signaling under endoplasmic reticulum stress, and chlorogenic acid protected pancreatic Β cells by enhancing the expression of insulin receptor substrate 2 via cAMP response element-binding protein and promoting insulin signaling downstream of insulin receptor substrate 2. In addition, chlorogenic acid was a potent antioxidant for the protection of pancreatic Β cells. Furthermore, in vivo and in vitro analyses revealed that the pancreatic Β cell-protective effect of chlorogenic acid was mediated by the alleviation of endoplasmic reticulum stress. The results suggest that these components of coffee have the potential to reduce the pathogenesis of type 2 diabetes and improve pancreatic Β cell insufficiency.

Obesity and risk for its comorbidities diabetes, hypertension, and dyslipidemia in Japanese individuals aged 65 years.

Diabetes, hypertension, and dyslipidemia are obesity-related comorbidities that contribute to the development of cardiovascular disease, one of the leading causes of death. In addition to obesity, the underweight condition is a concern because it can give rise to sarcopenia, particularly after the age of 65 years. We examined the risk for diabetes, hypertension, and dyslipidemia due to obesity in individuals of this age. We retrospectively investigated the relation between obesity and its three major comorbidities in 10,852 individuals aged 65 years who underwent health checkups implemented by Kobe City between April 2017 and March 2021. The prevalence of diabetes, hypertension, and dyslipidemia with and without hyper-low-density lipoprotein-cholesterolemia was 9.7%, 41.0%, 63.8%, and 19.5%, respectively, and the prevalence of these conditions increased with increasing obesity. The risk for diabetes and hypertension was increased markedly (odds ratios of 12.95 and 19.44, respectively), and that for dyslipidemia with and without hyper-low-density lipoprotein-cholesterolemia was modestly increased (odds ratios of 2.59 and 3.65, respectively) at a BMI of ≥ 35 kg/m2 compared with normal weight. Analysis by gender revealed that the obesity-related risk for dyslipidemia with hyper-low-density lipoprotein-cholesterolemia was small compared with other comorbidities in women, while the risk for all comorbidities elevated similarly in men. Our results suggest the importance of public health intervention for obesity to suppress its comorbidities, especially diabetes and hypertension, at this age.

l-Asparaginase regulates mTORC1 activity via a TSC2-dependent pathway in pancreatic beta cells.

Eif2ak4, a susceptibility gene for type 2 diabetes, encodes GCN2, a molecule activated by amino acid deficiency. Mutations or deletions in GCN2 in pancreatic ß-cells increase mTORC1 activity by decreasing Sestrin2 expression in a TSC2-independent manner. In this study, we searched for molecules downstream of GCN2 that suppress mTORC1 activity in a TSC2-dependent manner. To do so, we used a pull-down assay to identify molecules that competitively inhibit the binding of the T1462 phosphorylation site of TSC2 to 14-3-3. l-asparaginase was identified. Although l-asparaginase is frequently used as an anticancer drug for acute lymphoblastic leukemia, little is known about endogenous l-asparaginase. l-Asparaginase, which is expressed downstream of GCN2, was found to bind 14-3-3 and thereby to inhibit its binding to the T1462 phosphorylation site of TSC2 and contribute to TSC2 activation and mTORC1 inactivation upon TSC2 dephosphorylation. Further investigation of the regulation of mTORC1 activity in pancreatic ß-cells by l-asparaginase should help to elucidate the mechanism of diabetes and insulin secretion failure during anticancer drug use.

Preoperative Serum Cortisol Level Is Predictive of Weight Loss After Laparoscopic Sleeve Gastrectomy in Men with Severe Obesity but Not Women.

BACKGROUND: Bariatric surgery is an effective treatment for severe obesity and its associated medical problems. Preoperative factors that predict postoperative weight loss remain to be fully characterized, however. METHODS: Anthropometric and laboratory data were collected retrospectively for severely obese patients who underwent laparoscopic sleeve gastrectomy (LSG) between April 2016 and July 2019 at our hospital. Preoperative factors that predicted weight loss at 1 year after LSG were investigated. RESULTS: A total of 122 subjects (45 men and 77 women) underwent LSG. The mean ± SD age and body mass index at surgery were 44.4 ± 10.4 years and 40.7 ± 6.7 kg/m2. The percent total weight loss (%TWL) was 27.0 ± 8.6 among all subjects, 26.4 ± 8.0 among men, and 27.4 ± 8.9 among women, with no significant difference between the sexes. The %TWL showed a significant inverse correlation with serum cortisol level in men and with age and the visceral/subcutaneous fat area ratio in women. Multivariable regression analysis revealed the presence of type 2 diabetes and the serum cortisol concentration to be negatively associated with %TWL among all subjects and men, respectively. Receiver operating characteristic curve analysis identified an optimal cutoff of 10 µg/dL for prediction of a %TWL of ≥ 25 in men by serum cortisol level. CONCLUSIONS: Serum cortisol concentration was identified as a predictor for postoperative weight loss in men. Our results may thus help inform the decision to perform LSG or more effective surgical procedures in men with severe obesity.

Histone deacetylase 6 regulates insulin signaling in pancreatic ß cells.

The reduction of pancreatic ß cell mass is one of the key factors for the onset of type 2 diabetes. Many reports have indicated that insulin signaling is important for type 2 diabetes, but the mechanism by which insulin signaling is altered in pancreatic ß cells remains unclear. This study was designed to examine the role of histone deacetylases (HDACs) in the regulation of insulin signaling in pancreatic ß cells. We found that insulin signaling was downregulated by inhibition of HDAC6. HDAC6 expression was specifically observed in pancreatic ß cells and was decreased in the pancreatic islets of a type 2 diabetes mouse model. When a mouse pancreatic ß cell line (MIN6 cells) was treated with palmitic acid to mimic the effect of a high-fat diet on pancreatic ß cells, HDAC6 was imported into the nucleus. These results suggest that HDAC6 plays an important role in the regulation of insulin signaling in pancreatic ß cells. Therefore, clarifying the regulation of insulin signaling by HDAC6 may be a valuable approach for the treatment of type 2 diabetes.

Quantitative Evaluation of Functional Instability Due to Anterior Cruciate Ligament Deficiency.

BACKGROUND: A safe and simple procedure to evaluate functional instability due to anterior cruciate ligament (ACL) deficiency (ACLD) has not been established. The angle of trunk backward tilting, which is known as a posture at risk for ACL injuries, could be used as a parameter to evaluate functional instability due to ACLD. PURPOSE: To measure the backward tilt angle of the trunk with participants standing upright on 1 leg and to investigate its usefulness to quantitatively evaluate functional instability due to ACLD. STUDY DESIGN: Cohort study (diagnosis); Level of evidence, 3. METHODS: Our cohort included 50 participants with unilateral ACLD and 40 participants with bilateral healthy knees. The trunk backward tilt (TBT) test was conducted as follows: the participant was asked to maximally tilt the trunk backward in a single-leg standing position, while forward tilt of the index leg was blocked with a custom-made device. The TBT angle was measured using a side-view photograph. Subjective knee instability during the test was recorded using a visual analog scale (VAS). The relative and absolute reliability of the TBT test were verified in a sample of healthy participants and those with ACLD, and comparisons between indicators were performed. Multiple regression analysis was performed with the injured/uninjured side ratio (I/U ratio) of the TBT angle as the dependent variable and the following independent variables: (1) I/U ratio of knee extension muscle strength, (2) I/U ratio of knee flexion muscle strength, (3) side-to-side difference (SSD) of the KT-1000 arthrometer measurement, (4) sex, and (5) SSD of the VAS score. RESULTS: The TBT test had high reliability among healthy participants and those with ACLD. The TBT angle was significantly smaller and the VAS score was significantly higher on the injured side compared with the uninjured side and with healthy knees (P < .001 for all). Among the independent variables, the SSD of the VAS score had a negative influence on the I/U ratio of the TBT angle (R 2 = 0.59; P < .001). CONCLUSION: The TBT test is a simple, safe, and reliable method for quantitatively evaluating functional instability due to ACLD under weightbearing conditions that reflect subjective knee instability. The test will provide an index of treatment outcomes and return to sports through additional objective measurements before and after ACL reconstruction.

GCN2 regulates pancreatic ß cell mass by sensing intracellular amino acid levels.

EIF2AK4, which encodes the amino acid deficiency-sensing protein GCN2, has been implicated as a susceptibility gene for type 2 diabetes in the Japanese population. However, the mechanism by which GCN2 affects glucose homeostasis is unclear. Here, we show that insulin secretion is reduced in individuals harboring the risk allele of EIF2AK4 and that maintenance of GCN2-deficient mice on a high-fat diet results in a loss of pancreatic ß cell mass. Our data suggest that GCN2 senses amino acid deficiency in ß cells and limits signaling by mechanistic target of rapamycin complex 1 to prevent ß cell failure during the consumption of a high-fat diet.

Shoulder and elbow evaluation of pitchers in National High School Baseball Invitational Tournaments and National High School Baseball Championships from 1993 to 2016 in Japan.

BACKGROUND: Shoulder and elbow examinations for pitchers have been performed in Japan's National High School Baseball Invitational Tournaments (NHSBITs) and National High School Baseball Championships (NHSBCs) since 1993. However, for years the results have not been analyzed. The purpose of this study was to evaluate changes in the condition of shoulders and/or elbows of pitchers from 1993 to 2016. MATERIALS AND METHODS: Pitchers in NHSBITs and NHSBCs, 1994-2016, were examined together with those who received a trial examination (Trial) in the NHSBC 1993. Shoulder and elbow symptoms were comprehensively graded into five categories; none, mild, moderate, severe and dysfunction. Standard plain radiographs of the shoulder and elbow were obtained. Average Annual Percent Changes (AAPCs) in the percentages of pitchers with symptoms and positive radiographic findings were analyzed. RESULTS: The percentages of pitchers with moderate or worse symptoms in the shoulder or elbow were 14.6% and 13.8%, respectively, in the Trial, 1.1% and 1.1% in NHSBITs and 1.3% and 2.0% in NHSBCs. The AAPC of pitchers with a symptomatic shoulder in NHSBITs was reduced over the study period, at -3.36% (P < 0.05), but that in NHSBCs was unchanged at -1.01%. The AAPCs of pitchers with a symptomatic elbow in NHSBITs and in NHSBCs decreased, at -3.13% and -3.33%, respectively (P < 0.05), while that of pitchers with residual apophyseal fragmentation at the ulnar collateral ligament insertion increased at +2.79% (P < 0.05). The decreased percentages of symptomatic pitchers suggest that joint condition is well controlled in high school days; however, the increased frequency of radiographic findings suggests the necessity of protection against overuse in younger players. CONCLUSIONS: The percentages of pitchers with symptomatic shoulders and elbows in NHSBITs and those with symptomatic elbows in NHSBCs have decreased over the 23 years. However, the increased frequency of residual medial humeral epicondyle apophyseopathy should be noted.

Casein kinase 2 phosphorylates and stabilizes C/EBPß in pancreatic ß cells.

During the development of type 2 diabetes, endoplasmic reticulum (ER) stress leads to pancreatic ß cell failure. CCAAT/enhancer-binding protein (C/EBP) ß is highly induced by ER stress and AMP-activated protein kinase (AMPK) suppression in pancreatic ß cells, and its accumulation reduces pancreatic ß cell mass. We investigated the phosphorylation state of C/EBPß under these conditions. Casein kinase 2 (CK2) was found to co-localize with C/EBPß in MIN6 cells. It phosphorylated S222 of C/EBPß, a previously unidentified phosphorylation site. We found that C/EBPß is phosphorylated by CK2 under AMPK suppression and ER stress, which are important from the viewpoint of the worsening pathological condition of type 2 diabetes, such as decreased insulin secretion and apoptosis of pancreatic ß cells.

Compensatory hyperinsulinemia in high-fat diet-induced obese mice is associated with enhanced insulin translation in islets.

A high-fat diet (HF) is associated with obesity, insulin resistance, and hyperglycemia. Animal studies have shown compensatory mechanisms in pancreatic ß-cells after high fat load, such as increased pancreatic ß-cell mass, enhanced insulin secretion, and exocytosis. However, the effects of high fat intake on insulin synthesis are obscure. Here, we investigated whether insulin synthesis was altered in correlation with an HF diet, for the purpose of obtaining further understanding of the compensatory mechanisms in pancreatic ß-cells. Mice fed an HF diet are obese, insulin resistant, hyperinsulinemic, and glucose intolerant. In islets of mice fed an HF diet, more storage of insulin was identified. We analyzed insulin translation in mouse islets, as well as in INS-1 cells, using non-radioisotope chemicals. We found that insulin translational levels were significantly increased in islets of mice fed an HF diet to meet systemic demand, without altering its transcriptional levels. Our data showed that not only increased pancreatic ß-cell mass and insulin secretion but also elevated insulin translation is the major compensatory mechanism of pancreatic ß-cells.

Pancreatic ß-cell failure mediated by mTORC1 hyperactivity and autophagic impairment.

Hyperactivation of the mammalian target of rapamycin complex 1 (mTORC1) in ß-cells is usually found as a consequence of increased metabolic load. Although it plays an essential role in ß-cell compensatory mechanisms, mTORC1 negatively regulates autophagy. Using a mouse model with ß-cell-specific deletion of Tsc2 (ßTsc2(-/-)) and, consequently, mTORC1 hyperactivation, we focused on the role that chronic mTORC1 hyperactivation might have on ß-cell failure. mTORC1 hyperactivation drove an early increase in ß-cell mass that later declined, triggering hyperglycemia. Apoptosis and endoplasmic reticulum stress markers were found in islets of older ßTsc2(-/-) mice as well as accumulation of p62/SQSTM1 and an impaired autophagic response. Mitochondrial mass was increased in ß-cells of ßTsc2(-/-) mice, but mitophagy was also impaired under these circumstances. We provide evidence of ß-cell autophagy impairment as a link between mTORC1 hyperactivation and mitochondrial dysfunction that probably contributes to ß-cell failure.

Ablation of TSC2 enhances insulin secretion by increasing the number of mitochondria through activation of mTORC1.

AIM: We previously found that chronic tuberous sclerosis protein 2 (TSC2) deletion induces activation of mammalian target of rapamycin Complex 1 (mTORC1) and leads to hypertrophy of pancreatic beta cells from pancreatic beta cell-specific TSC2 knockout (ßTSC2(-/-)) mice. The present study examines the effects of TSC2 ablation on insulin secretion from pancreatic beta cells. METHODS: Isolated islets from ßTSC2(-/-) mice and TSC2 knockdown insulin 1 (INS-1) insulinoma cells treated with small interfering ribonucleic acid were used to investigate insulin secretion, ATP content and the expression of mitochondrial genes. RESULTS: Activation of mTORC1 increased mitochondrial DNA expression, mitochondrial density and ATP production in pancreatic beta cells of ßTSC2(-/-) mice. In TSC2 knockdown INS-1 cells, mitochondrial DNA expression, mitochondrial density and ATP production were increased compared with those in control INS-1 cells, consistent with the phenotype of ßTSC2(-/-) mice. TSC2 knockdown INS-1 cells also exhibited augmented insulin secretory response to glucose. Rapamycin inhibited mitochondrial DNA expression and ATP production as well as insulin secretion in response to glucose. Thus, ßTSC2(-/-) mice exhibit hyperinsulinemia due to an increase in the number of mitochondria as well as enlargement of individual beta cells via activation of mTORC1. CONCLUSION: Activation of mTORC1 by TSC2 ablation increases mitochondrial biogenesis and enhances insulin secretion from pancreatic beta cells.

Ablation of C/EBPbeta alleviates ER stress and pancreatic beta cell failure through the GRP78 chaperone in mice.

Pancreatic beta cell failure is thought to underlie the progression from glucose intolerance to overt diabetes, and ER stress is implicated in such beta cell dysfunction. We have now shown that the transcription factor CCAAT/enhancer-binding protein beta (C/EBPbeta) accumulated in the islets of diabetic animal models as a result of ER stress before the onset of hyperglycemia. Transgenic overexpression of C/EBPbeta specifically in beta cells of mice reduced beta cell mass and lowered plasma insulin levels, resulting in the development of diabetes. Conversely, genetic ablation of C/EBPbeta in the beta cells of mouse models of diabetes, including Akita mice, which harbor a heterozygous mutation in Ins2 (Ins2WT/C96Y), and leptin receptor-deficient (Lepr-/-) mice, resulted in an increase in beta cell mass and ameliorated hyperglycemia. The accumulation of C/EBPbeta in pancreatic beta cells reduced the abundance of the molecular chaperone glucose-regulated protein of 78 kDa (GRP78) as a result of suppression of the transactivation activity of the transcription factor ATF6alpha, thereby increasing the vulnerability of these cells to excess ER stress. Our results thus indicate that the accumulation of C/EBPbeta in pancreatic beta cells contributes to beta cell failure in mice by enhancing susceptibility to ER stress.

Effect of intrauterine undernutrition during late gestation on pancreatic beta cell mass.

We analyzed the effect of low birth weight on pancreatic beta cell mass. We used pregnant C57BL6J mice, and we reduced their food supply by 30% during the late gestational period and examined the changes in the metabolism and pancreatic beta cell mass. Pancreatic beta cell mass at birth was greatly decreased in the mice of the food restriction group (RG) as compared to the mice of the control group (CG). The body weight of RG mice exhibited a "catch-up growth" pattern and became equivalent to that of CG mice 7 days after birth, and thereafter exceeded that of CG mice; however, the pancreatic beta cell mass in RG mice remained lower than that in CG mice at the age of 4 weeks. A high-fat diet significantly increased the pancreatic beta cell mass in RG mice as compared to that in CG mice at 12 weeks of age. However, RG mice fed on high-fat diets tended to exhibit a decrease in the pancreatic beta cell mass at approximately 20 weeks of age. The plasma insulin concentrations also tended to be decreased in RG mice after 24 weeks of age as compared to those of CG mice. These results thus indicate that the growth of pancreatic beta cells is insufficient in RG mice, and pancreatic beta cell failure can easily develop as a consequence of insulin resistance.

Asymmetric load-carrying in young and elderly women: relationship with lower limb coordination.

This study investigates balance during asymmetric load-carrying and how asymmetric loading affects lower limb coordination during gait. Walking with and without a hand-held bag was analyzed in five young and six elderly women using a 6-camera VICON system and two force plates. Balance and lower limb coordination were compared when walking with and without a bag and also between age groups. While carrying a bag, the trunk, head, and upper arm were recruited in both young and elderly women. With the load, the contralateral hip abduction torque increased, whereas the ipsilateral hip torque decreased. Intralimb and interlimb coordinations did not vary with the different load conditions. The only difference observed between the groups was the contralateral shoulder abduction. The elderly group abducted their shoulders to a greater extent, even when walking without a bag.

Arthroscopic anterior cruciate ligament reconstruction using fresh-frozen bone plug-free allogeneic tendons: 10-year follow-up.

PURPOSE: To evaluate the long-term outcomes following arthroscopic-assisted anterior cruciate ligament (ACL) reconstruction using fresh-frozen allogeneic tendon. METHODS: Sixty-one athletically active patients (mean age at surgery, 20.9 years) who had arthroscopic-assisted ACL reconstruction using fresh-frozen free tendon allograft underwent physical examination, instrumented laxity measurement, radiographs, and thigh muscle strength at 10 to 14 years (mean, 11.5) postoperatively, and these data were compared with results of similar measurements obtained at 2 years postoperatively. RESULTS: Lachman test and pivot shift test were maintained as negative in 53 (87%) and 52 (85%) patients at follow-up, respectively. Quantitative measurements using the KT-2000 knee arthrometer showed 1.6 +/- 1.3 (mean +/- SD) mm in the side-to-side difference, and no more than 3 mm in 56 patients (92%) at final follow-up. All but one was assessed as normal or nearly normal by International Knee Documentation Committee score. Patient activity level was decreased at the long-term follow-up, but this change was mainly associated with changes in social context (e.g., graduation from school) rather than with knee limitations. None of the patients experienced deep infection or graft rejection. Degenerative joint disease on radiographs was seen in 13 out of 15 patients (87%) whose menisci had been excised, whereas it was observed in 12 out of 46 (26%) whose menisci had been preserved. CONCLUSIONS: A series of 61 patients with fresh-frozen bone plug-free tendon allografts using a 2-incision technique for arthroscopic ACL reconstruction results in long-term knee stabilization and functioning among young active individuals while simultaneously avoiding graft harvest site morbidity. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Biphasic response of pancreatic beta-cell mass to ablation of tuberous sclerosis complex 2 in mice.

Recent studies have demonstrated the importance of insulin or insulin-like growth factor 1 (IGF-1) for regulation of pancreatic beta-cell mass. Given the role of tuberous sclerosis complex 2 (TSC2) as an upstream molecule of mTOR (mammalian target of rapamycin), we examined the effect of TSC2 deficiency on beta-cell function. Here, we show that mice deficient in TSC2, specifically in pancreatic beta cells (betaTSC2(-/-) mice), manifest increased IGF-1-dependent phosphorylation of p70 S6 kinase and 4E-BP1 in islets as well as an initial increased islet mass attributable in large part to increases in the sizes of individual beta cells. These mice also exhibit hypoglycemia and hyperinsulinemia at young ages (4 to 28 weeks). After 40 weeks of age, however, the betaTSC2(-/-) mice develop progressive hyperglycemia and hypoinsulinemia accompanied by a reduction in islet mass due predominantly to a decrease in the number of beta cells. These results thus indicate that TSC2 regulates pancreatic beta-cell mass in a biphasic manner.