Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Pregnancy-associated osteoporosis: Seven cases of pregnancy-associated osteoporosis discussed

WOS: 000088348100345

Fibromyalgia, physical and psychological aspects: 107 cases

Fibromyalgia syndrome which is characterized by, widespread musculoskeletal pain, sleep disturbance, fatigue, morning stiffness and tenderness at specific sites is frequently associated with affective dysfunction. The aim of this study is to evaluate patients with fibromyalgia both physically and psychologically. Patients with fibromyalgia syndrome, who were admitted to Cukurova University Faculty of Medicine, Department of Physical Medicine and Rehabilitation are studied. Of the 107 patients, 96 were female and 11 male. In all patients, tender points, factors modulating the symptoms, associated features, laboratory and radiologic findings were determined. We performed Turkish version of Maudsley Obsessional-Compulsive Questionnaire and Zung's Self Rating Depression Scale in patients to evaluate them psychologically. In our study group, the number of tender points varied between 11 and 18. We observed the folio wing associated disturbances: sleep disturbance in 65%, fatigue in 93%, morning stiffness in 72%, subjective paresthesias in 75%, subjective soft tissue swelling in 45%, anxiety in 57%, headache in 73%, irritable bowel syndrome in 43%, widespread pain in 42%, sicca symptoms in 18%, urinary urgency in 24% and dysmenorrhea in 23% (rates). The psychological evaluation of the patients revealed that 43% of them had depressive symptoms and 25% had obsessive-compulsive sings. In conclusion we demonstrated that, besides physical findings, fibromyalgia syndrome is also associated with several psychological symptoms

Reduction of spinal cord injury with pentobarbital and hypothermia in a rabbit model

WOS: 000179554600013PubMed ID: 12443752Objectives: to evaluate the effects of hypothermia and pentobarbital on spinal cord ischaemia induced in a rabbit model. Materials and Methods: thirty-two rabbits, allocated into four equal groups, had the infrarenal aorta clamped distal to the left renal artery and above the iliac bifurcation for 40 min. Groups 3 and 4 had infusion of 15 mg/kg of pentobarbital intravenously for 5 min, 15 min before the cross-clamping. Groups 2 and 4 had infusion of 20 ml of Ringer's lactate (LR) solution at 3degreesC for 3 min during aortic cross clamp into the isolated aortic segment. Group 1 was untreated and served as control. Postoperative functions of spinal cord were assessed. Results: paraplegia occurred in all rabbits in Group 1, in one in each of Groups 2 and 3, whereas no paraplegia was observed in Group 4. In addition 2 and 3 animals of Groups 2 and 3, respectively revealed varying degree of neurological disturbances, whereas all animals of Group 4 had normal function. This difference between Groups 2, 3, and 4 vs Group 1 was significant (p < 0.002). So was the difference between Groups 2 and 4 (p = 0.03), whereas the difference between Groups 3 and 4 was not significant. Conclusions: hypothermia and pentobarbital was more effective than hypothermia alone for prevention of spinal cord ischaemia in a rabbit model

Double crush syndrome: Presentation of 5 cases

Double crush syndrome is a general term referring to the coexistence of dual compressive lesions along the course of a nerve. We present 5 cases of double-crush syndrome and discuss the clinical evidence, symptomatoloy, findings of this condition. Persistence of symptoms after initial treatment of a nerve entrapment is an indication to search for another site of compression. However it is more important to identify before any surgical intervention is planned, those patients who may have double crush lesions and direct the treatment toward both processes

Primary tubular choriorentinitis

The aim of this report is to emphasize that corticosteroids should not be prescribed when the etiological factors remain unidentified. A 34‑year‑old male visited our ophthalmology clinic suffering from blurred vision. Behçet’s disease had been diagnosed 5 years prior, and the patient was taking an oral immunosuppressant and a systemic corticosteroid. Vitreous cell count and foci of chorioretinitis (apparently confined to the ocular fundus) were evident. A vitreous sample was subjected to polymerase chain reaction, which resulted in the identification of and Mycobacterium tuberculosis. Treatment with a combination of four anti‑tuberculosis drugs was commenced. During the follow‑up, the vitritis disappeared, and the foci of active chorioretinitis improved. Systemic and Sub‑Tenon corticosteroid treatment should not be prescribed until vitreous and anterior chamber fluid samples have been evaluated in patients with uveitis. Some microbes are not detected on routine examination.Keywords: Corticosteroid therapy, Tuberculosis, Chorioretinitis, Uveiti

Clinical and echocardiographic results of the Kalangos biodegradable tricuspid ring for moderate and severe functional tricuspid regurgitation treatment.

INTRODUCTION: The Kalangos Biodegradable Tricuspid Ring (Kalangos Biodegradable Tricuspid Ring®, Bioring SA, Lonay, Switzerland) is a biodegradable prothesis in the treatment of functional tricuspid regurgitation (FTR). In this study, we aimed to determine the clinical and echocardiographic results of this prosthesis for moderate and severe FTR treatment and compare this technique with the results of semicircular DeVega annuloplasty. MATERIALS AND METHODS: From January 2005 through January 2010 we retrospectively studied the data on 64 consecutive patients underwent annuloplasty procedures for FTR treatment. The patients were assigned to 2 groups: (1) Kalangos BTR annuloplasty was performed in 32 patients, and (2) Semicircular DeVega annuloplasty was performed in the 32 patients. All patients were evaluated clinically and by echocardiography preoperatively, at the end of the 1st week, 3rd and 6th month following surgery. RESULTS: No complications related to the prosthesis or the procedures within the follow-up period were recorded. At the follow-up period, systolic pulmonary arterial pressure and tricuspid valve area diameter were found to be significantly lower than the preoperative values in both groups (p < 0.0001). At the follow-up period residual tricuspid regurgitation and the Tei index (Myocardial performance index) was significantly lower in group 1 compared to group 2 (p < 0.05). Three-quarters of the annuloplasty ring had degraded in the postoperative 6-months period. DISCUSSION: We conclude that Kalangos BTR is an efficient and safe prosthesis with easy implantation technique for FTR treatment, with encouraging midterm results