We report in this study the identification of a natural product-like antagonist () of Vps34 as a potent autophagy modulator structure-based virtual screening. Aurone derivative strongly inhibited Vps34 activity in cell-free and cell-based assays. Significantly, prevents autophagy in human cells induced either by starvation or by an mTOR inhibitor. modeling and kinetic data revealed that could function as an ATP-competitive inhibitor of Vps34. Moreover, it suppressed autophagy and without inducing heart or liver damage in mice. could be utilized as a new motif for more selective and efficacious antagonists of Vps34 for the potential treatment of autophagy-related human diseases.

BACKGROUNDSeveral reports have shown the progression of articular cartilage degeneration after anterior cruciate ligament (ACL) reconstruction. No report has been published about the cartilage comparing changes after single-bundle (SB) and double-bundle (DB) ACL reconstructions. The purpose of this study was to evaluate the articular cartilage changes after SB and DB ACL reconstructions by second-look arthroscopy.

METHODSNinety-nine patients who received arthroscopic ACL reconstruction were retrospectively reviewed at an average of 14 months after reconstruction, 58 patients underwent SB ACL reconstruction and 41 patients underwent DB ACL reconstruction. Hamstring tendon autografts were used in all patients. Second-look arthroscopy was done in conjunction with the tibial staple fixation removal at least one year after the initial ACL reconstruction. Arthroscopic evaluation and grading of the articular cartilage degeneration for all patients were performed at the initial ACL reconstruction, and at the second-look arthroscopy.

RESULTSThe average cartilage degeneration at the patellofemoral joint (PFJ) was found significantly worsened after both SB and DB ACL reconstructions. This worsening were not seen at medial tibiofemoral joint (TFJ) and lateral TFJ. Grade II cartilage damage was the most common. At second-look arthroscopy, the average patellar cartilage degeneration was 1.14 ± 0.14 (at first look 0.52 ± 0.11) for the SB group, and 1.22 ± 0.15 (at first look 0.56 ± 0.12) for the DB group. The average trochlear cartilage degeneration was 1.05 ± 0.16 (at fist look 0.10 ± 0.06) and 0.66 ± 0.17 (at fist look 0.17 ± 0.09), respectively. The average patellar cartilage degeneration showed no significant difference in both groups. However, the average trochlea cartilage degeneration in DB group was significantly less than in SB group.

CONCLUSIONSPatellofemoral cartilage degeneration continued to aggravate after ACL reconstruction. DB ACL reconstruction could significantly decrease the trochlea cartilage degeneration compared with SB ACL reconstruction.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Reconstruction ; methods ; Arthroscopy ; methods ; Cartilage, Articular ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Second-Look Surgery ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo detect single nucleotide polymorphisms (SNPs) of the myocilin (MYOC) gene and to investigate their associations with primary open-angle glaucoma (POAG).

METHODSOne hundred and fifty-seven sporadic patients with POAG and 155 unrelated control subjects without POAG were recruited from staff and visitors to the Prince of Wales Hospital between 1998 and 2000. All study subjects are ethnic Chinese living in Hong Kong. The two populations were matched in frequencies of gender and age. The SNPs of the MYOC gene in POAG patients and control subjects were screened and identified by high throughout conformation sensitive gel electrophoresis and fluorescent labeling automated sequencing. The genotype frequencies of each SNP in the two groups were compared by the Chi2 test or Fisher's exact 2-tailed test.

RESULTSA total of seventeen SNPs were identified from 2172 bp long of the MYOC gene, including all 3 exons and adjacent non-coding regions. The identified SNPs were 1-83G --> A, G12R, P16L, A17S, R46X, R76K, R91X, T123T, D208E, L215P, 730+35A --> G, A260A, I288I, E300K, T353I, Y471C and 1515+73G --> C, respectively. Of these, R91X, E300K and Y471C were found only in POAG patients. A significant difference between POAG patients and control subjects was found in the genotype frequencies of 1515+73G --> C. The frequency of the heterozygote (CG) was 0.6% in POAG patients, significantly less than the 4.5% in control subjects (Fisher's exact 2-tailed test, P=0.036, OR=0.136, 95%CI=0.022-0.828). No significant difference was found between the two populations in genotype frequencies of all other SNPs.

CONCLUSIONThe polymorphisms of the MYOC gene may be related to POAG.

Adult ; Aged ; Aged, 80 and over ; Amino Acid Substitution ; Base Sequence ; Case-Control Studies ; Cytoskeletal Proteins ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Eye Proteins ; genetics ; Female ; Gene Frequency ; Genotype ; Glaucoma, Open-Angle ; genetics ; pathology ; Glycoproteins ; genetics ; Humans ; Male ; Middle Aged ; Point Mutation ; Polymorphism, Single Nucleotide

BACKGROUNDPrimary open-angle glaucoma (POAG) is characterized by optic nerve damage and consists of a group of genetically heterogeneous disorders. This study was to investigate the associations of genetic and environmental factors with POAG in a hospital-based Chinese population.

METHODSThirty-two adult onset POAG patients and 96 age-sex matched control subjects were studied by multivariable logistic regression analysis for the relationships between POAG and its risk factors including family history, diabetes, hypertension, cardiovascular diseases, cigarette smoking, alcohol consumption and polymorphisms of the myocilin and the optineurin genes.

RESULTSUnivariate analysis showed that POAG was related to family history, cardiovascular disease, alcohol consumption and a myocilin sequence alteration (T353I) (P < 0.04). Multivariable logistic regression analysis confirmed that POAG was significantly associated with family history (OR = 20.2), hypertension (OR = 3.58), cigarette smoking (OR = 10.8), alcohol consumption (OR = 0.028) and T353I (OR = 6.03, all P < 0.05).

CONCLUSIONSFamily history, hypertension, cigarette smoking and T353I in the myocilin gene are risk factors for POAG. Alcohol consumption, however, has a protective effect.

Adult ; Aged ; Aged, 80 and over ; Alcohol Drinking ; adverse effects ; Cytoskeletal Proteins ; Eye Proteins ; genetics ; Female ; Glaucoma, Open-Angle ; etiology ; genetics ; Glycoproteins ; genetics ; Humans ; Hypertension ; complications ; Logistic Models ; Male ; Middle Aged ; Risk Factors ; Smoking ; adverse effects

To demonstrate the age-related changes in the dynamics of the nociceptive discharge of dorsal horn nociceptive neurons, the nonlinear prediction method was used to quantify the degree of deterministic behavior within the interspike interval series of tissue injury-induced firing of spinal nociceptive neurons in anesthetized adult young (3-4 months) and aged (>22 months) rats. Subcutaneous bee venom injection induced long-term discharge of spinal wide dynamic range (WDR) neurons in both groups. However, the nociceptive discharge of single WDR neurons in the aged group showed higher determinism when compared with the adult young rats. This result suggests that the dynamics of single nociceptive neurons may not remain constant throughout the life span, and this age-associated change may be an underlying mechanism for various pain manifestations in the elderly population.
Age Factors ; Aging ; Animals ; Bee Venoms ; Behavior, Animal ; Electrophysiology ; Nociceptors ; physiopathology ; Pain ; physiopathology ; Pain Threshold ; Posterior Horn Cells ; physiopathology ; Rats ; Rats, Sprague-Dawley