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  1. Singla H, Alexander M
    J Maxillofac Oral Surg, 2015 Jun;14(2):423-31.
    PMID: 26028869 DOI: 10.1007/s12663-014-0648-2
    Local anesthesia has been a boon for dentistry to allay the most common fear of pain among dental patients. Several techniques to achieve anesthesia for posterior maxillae have been advocated albeit with minor differences. We compared two techniques of posterior superior alveolar nerve block (PSANB), the one claimed to be "most accurate" to the one "most commonly used."
  2. Sharma SD, Vidya B, Alexander M, Deshmukh S
    J Maxillofac Oral Surg, 2015 Sep;14(3):611-5.
    PMID: 26225052 DOI: 10.1007/s12663-014-0723-8
    The aim of this study was to evaluate the efficacy of periotomes in single rooted nonsurgical tooth extractions.
  3. Kumar S, Thomas BS, Gupta K, Guddattu V, Alexander M
    Niger J Clin Pract, 2018 Aug;21(8):1029-1033.
    PMID: 30074006 DOI: 10.4103/njcp.njcp_341_17
    Aim and objectives: The aim and the objectives were. (1) to assess the efficacy of a desensitizing toothpaste containing 8.0% arginine-calcium carbonate (Colgate® Sensitive Pro-Relief™), (2) to assess the efficacy of a desensitizing toothpaste containing 8.0% arginine-calcium carbonate (Colgate® Sensitive Pro-Relief™) used in combination with iontophoresis, and (3) to compare the effectiveness of the above methods.

    Subjects and Methods: Two groups of 40 patients each having dentinal hypersensitivity were treated using 8% proarginine and iontophoresis. The patients were recalled after 1, 2, and 4 weeks. The scores were tabulated and the results were analyzed using SPSS statistical software.

    Results: Visual analog scale between the two groups showed a significant difference from the 1st week till the 4th week. ANOVA values showed the reduction in the dentinal hypersensitivity in Group 2 using the iontophoresis along with the 8.0% arginine-calcium carbonate toothpaste. The Cochran-Mantel-Haenszel correlation test of the Schiff's dentinal hypersensitivity cross-tabulation showed P < 0.001 which was statistically significant reduction after the 4th week following the application of 8.0% arginine-calcium carbonate along with iontophoresis.

    Conclusion: Iontophoresis, when used along with Colgate® Sensitive Pro-Relief™ toothpaste, can provide additional benefit as this provides a better sealing effect.

  4. Sharma SD, Gupta A, Bansal P, Alexander M, Vidya B, Gupta H
    Natl J Maxillofac Surg, 2022 Aug;13(Suppl 1):S91-S96.
    PMID: 36393928 DOI: 10.4103/njms.njms_309_21
    Aim: The goal of this study was to assess the effectiveness of piezotome as compared to periotome extractions of nonrestorable endodontic treatment of teeth in terms of operational time, pain control, and postoperative bone loss considering the prosthetic rehabilitation in future.

    Materials and Methods: A double-blind, randomized controlled trial was conducted with 100 patients who wanted single-rooted teeth to be extracted (which failed endodontically). The participants have been randomized into two equal groups named as - (i) a periotome group (ii) and a piezotome group. Duration of the surgery, postoperative pain within 7 days, complications (if any) associated with the extraction process were performed as a part of clinical assessment. Bone loss has been analyzed 6 months after the surgery radiographically. The data have been recorded and analyzed using the version 22.0 of the SPSS software package.

    Results: All parameters in the periotome category (P < 0.05) were statistically significant except for bone loss and gingival laceration in comparison to piezotome group. In the piezotome group, a longer time was observed for surgery and delayed pain control was achieved. In our study, we found statistically significant more marginal bone loss in piezotome group in comparison with periotome group.

    Conclusion: The findings of this study indicate that for intraoperative and postoperative comfort periotome could be used as a safer and cheaper option for atraumatic extractions but piezosurgery may prove as a better choice soon for surgeries in the maxillofacial region to maintain soft-tissue integrity.

  5. Garg R, Mishra N, Alexander M, Gupta SK
    Ann Maxillofac Surg, 2017 Jul-Dec;7(2):237-244.
    PMID: 29264292 DOI: 10.4103/ams.ams_87_17
    Introduction: With introduction of the term "ossteointegration of dental implant" by Branemark, advancement in implantology from 1957 to 2017 has come a long way with modification in implant type and in loading time. This study aims to evaluate the survival of endo-osseous immediate loading (IL) implant and basal IL implants in atrophic jaws with objective to compare implant survival in atrophic jaws for full mouth rehabilitation between endo-osseous IL versus endo-osseous delayed loading (DL) versus basal IL during 3-year follow-up.

    Materials and Methods: Fifty-two (34 endo-osseous and 18 basal) implants were placed in 4 patients requiring full mouth rehabilitation in atrophic jaws. Case 1: Endo-osseous DL implants in upper and lower arch, Case 2: Endo-osseous IL implants in upper and lower arch, Case 3: Basal IL implant in upper and lower arch, and Case 4: Endo-osseous DL in upper arch and basal IL implant in the lower arch. Intraoperative evaluation was done on the basis of pain (visual analog scale [VAS]), operative time, and initial primary implant stability. Postoperative evaluation was done on pain (VAS), infection, radiographically successful implant (orthopantomogram), and patient satisfaction (Grade 0-10).

    Results: All cases showed satisfactory results but more amount of intra- and post-operative pain was felt with immediate basal implants.

    Conclusion: We believe that clinicians should comply with patient requests, and for this reason, we agree with some authors to use minimally invasive techniques and to avoid when possible esthetic or functional problems associated with the use of removable prosthesis after teeth extractions.
  6. Payne KFB, Higginson J, Basyuni S, Goodson AMC, Chadha A, Elledge R, et al.
    Br J Oral Maxillofac Surg, 2023 Feb;61(2):124-130.
    PMID: 36774281 DOI: 10.1016/j.bjoms.2023.01.003
    Entering into surgical academia can seem a daunting prospect for an oral and maxillofacial surgery (OMFS) trainee. However, the streamlining of academic training by the NIHR to create the integrated academic training (IAT) pathway has simplified academic training and more clearly defined academic positions and entry points for trainees. In this article we review the current NIHR IAT pathway and the various grades and entry points available to OMF surgeons, both pre- and post-doctoral. We highlight the unique challenges facing OMF trainees and provide advice and insight from both junior and senior OMFS academics. Finally, we focus on the planning and application for a doctoral research fellowship - discussing funding streams available to OMF surgeons.
  7. Stuart AM, Merfield CN, Horgan FG, Willis S, Watts MA, Ramírez-Muñoz F, et al.
    Environ Sci Pollut Res Int, 2023 Feb;30(7):16984-17008.
    PMID: 36622585 DOI: 10.1007/s11356-022-24951-0
    A small proportion of the thousands of pesticides on the market today are associated with a disproportionately high incidence of severe acute pesticide poisoning and suicide. Paraquat stands out as one of the most lethal pesticides in common use, frequently involved in fatal incidents due to suicides or accidental exposure. Even though paraquat has been banned in over 67 countries, it is still widely used in many others, particularly in Asia and Latin America. Based on a literature review and consultations, this paper identifies options for replacing paraquat and distils practical lessons from numerous successes around the world. Our aim is to support regulators, policymakers, agronomists and the supply chain sector with practical information related to phasing out paraquat. Production data consistently failed to show any negative effects of banning paraquat on agricultural productivity. A wide range of alternative approaches to weed management and crop defoliation are available, many of which do not rely on herbicides. Over 1.25 million farmers in low- and middle-income countries (LMICs) successfully produce a range of crops for private voluntary standards (PVS) in food and fiber supply chains which prohibit paraquat use. We conclude from the findings of this study that eliminating paraquat will save lives without reducing agricultural productivity. Less hazardous and more sustainable alternatives exist. To enhance successful adoption and uptake of these methods on a wide scale, farmers require training and support within an enabling policy environment.
  8. Alexander MJ, Sinnatamby AS, Rohaimah MJ, Harun AH, Ng JS
    Ann Acad Med Singap, 1990 May;19(3):344-6.
    PMID: 2393233
    Brunei Darussalam has a mixed population with entirely different cultures and religions. The overall incidence of Hepatitis B virus (HBV) infection is 6%. A racial analysis of the incidence of HBV infection in Brunei shows a significantly higher incidence in Chinese compared to the other races. This is consistent with the incidence in the neighbouring countries.
  9. Tan JH, Sivadurai G, Tan HCL, Tan YR, Jahit S, Hans Alexander M
    Surg Laparosc Endosc Percutan Tech, 2020 Apr;30(2):106-110.
    PMID: 31923160 DOI: 10.1097/SLE.0000000000000754
    BACKGROUND: Provision of enteral nutrition with jejunal feeding in upper gastrointestinal obstruction is highly recommended. Access to jejunum can be obtained surgically, percutaneously, or endoscopically. Our institution routinely and preferentially utilizes a silicone nasojejunal tube that is inserted past the obstruction endoscopically. We use a custom dual channel tube that allows feeding at the distal tip and another channel 40 cm from the tip that enables decompression proximally. This is a report of our experience with this custom nasojejunal tube.

    METHODS: This is a prospective observational study of 201 patients who underwent endoscopic nasojejunal wire-guided feeding tube insertions for obstruction of either the esophagus or the stomach including both benign and malignant pathologies between January 2015 to June 2018 in Hospital Sungai Buloh and Hospital Sultanah Aminah, Malaysia. The indications for tube insertion, insertion technique, and tube-related problems were described.

    RESULTS: The nasojejunal tube was used to establish enteral feeding in patients with obstructing tumors of the distal esophagus in 65 patients (32.3%) and gastric outlet obstruction in 72 patients (35.8%). There were 54 patients (26.9%) who required reinsertion. The most common reason for reinsertion was unintentional dislodgement, where 32 patients (15.9%) followed by tube blockage 20 patients (10.0%). Using our method of advancement under direct vision, we had only 2 cases of malposition due to severely deformed anatomy. We had no incidence of aspiration in this group of patients and overall, the patients tolerated the tube well.

    CONCLUSIONS: The novel nasojejunal feeding tube with gastric decompression function is a safe and effective method of delivery of enteral nutrition in patients with upper gastrointestinal obstruction. These tubes if inserted properly are well tolerated with almost no risk of malposition and are tolerated well even for prolonged periods of time until definitive surgery could be performed.

  10. Merican I, Guan R, Amarapuka D, Alexander MJ, Chutaputti A, Chien RN, et al.
    J Gastroenterol Hepatol, 2000 Dec;15(12):1356-61.
    PMID: 11197043
    Of the estimated 50 million new cases of hepatitis B virus (HBV) infection diagnosed annually, 5-10% of adults and up to 90% of infants will become chronically infected, 75% of these in Asia where hepatitis B is the leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC). In Indonesia, 4.6% of the population was positive for HBsAg in 1994 and of these, 21% were positive for HBeAg and 73% for anti-HBe; 44% and 45% of Indonesian patients with cirrhosis and HCC, respectively, were HBsAg positive. In the Philippines, there appear to be two types of age-specific HBsAg prevalence, suggesting different modes of transmission. In Thailand, 8-10% of males and 6-8% of females are HBsAg positive, with HBsAg also found in 30% of patients with cirrhosis and 50-75% of those with HCC. In Taiwan, 75-80% of patients with chronic liver disease are HBsAg positive, and HBsAg is found in 34% and 72% of patients with cirrhosis and HCC, respectively. In China, 73% of patients with chronic hepatitis and 78% and 71% of those with cirrhosis and HCC, respectively, are HBsAg positive. In Singapore, the prevalence of HBsAg has dropped since the introduction of HBV vaccination and the HBsAg seroprevalence of unvaccinated individuals over 5 years of age is 4.5%. In Malaysia, 5.24% of healthy volunteers, with a mean age of 34 years, were positive for HBsAg in 1997. In the highly endemic countries in Asia, the majority of infections are contracted postnatally or perinatally. Three phases of chronic HBV infection are recognized: phase 1 patients are HBeAg positive with high levels of virus in the serum and minimal hepatic inflammation; phase 2 patients have intermittent or continuous hepatitis of varying degrees of severity; phase 3 is the inactive phase during which viral concentrations are low and there is minimal inflammatory activity in the liver. In general, patients who clear HBeAg have a better prognosis than patients who remain HBeAg-positive for prolonged periods of time. The outcome after anti-HBe seroconversion depends on the degree of pre-existing liver damage and any subsequent HBV reactivation. Without pre-existing cirrhosis, there may be only slight fibrosis or mild chronic hepatitis, but with pre-existing cirrhosis, further complications may ensue. HBsAg-negative chronic hepatitis B is a phase of chronic HBV infection during which a mutation arises resulting in the inability of the virus to produce HBeAg. Such patients tend to have more severe liver disease and run a more rapidly progressive course. The annual probability of developing cirrhosis varies from 0.1 to 1.0% depending on the duration of HBV replication, the severity of disease and the presence of concomitant infections or drugs. The annual incidence of hepatic decompensation in HBV-related cirrhosis varies from 2 to 10% and in these patients the 5-year survival rate drops dramatically to 14-35%. The annual risk of developing HCC in patients with cirrhosis varies between 1 and 6%; the overall reported annual detection rate of HCC in surveillance studies, which included individuals with chronic hepatitis B and cirrhosis, is 0.8-4.1%. Chronic hepatitis B is not a static disease and the natural history of the disease is affected by both viral and host factors. The prognosis is poor with decompensated cirrhosis and effective treatment options are limited. Prevention of HBV infection thorough vaccination is still, therefore, the best strategy for decreasing the incidence of hepatitis B-associated cirrhosis and HCC.
  11. Langford-Smith AWW, Hasan A, Weston R, Edwards N, Jones AM, Boulton AJM, et al.
    Sci Rep, 2019 02 19;9(1):2309.
    PMID: 30783159 DOI: 10.1038/s41598-019-38921-z
    Endothelial colony forming progenitor cell (ECFC) function is compromised in diabetes, leading to poor vascular endothelial repair, which contributes to impaired diabetic foot ulcer healing. We have generated novel glycomimetic drugs with protective effects against endothelial dysfunction. We investigated the effect of glycomimetic C3 on the functional capacity of diabetic ECFCs. ECFCs were isolated from healthy controls and patients with diabetes with neuroischaemic (NI) or neuropathic (NP) foot ulcers. Functionally, diabetic ECFCs demonstrated delayed colony formation (p 
  12. Hiew FL, Thit WM, Alexander M, Thirugnanam U, Siritho S, Tan K, et al.
    J Cent Nerv Syst Dis, 2021;13:11795735211057314.
    PMID: 35173510 DOI: 10.1177/11795735211057314
    Therapeutic plasma exchange (TPE) is an effective and affordable treatment option in most parts of Southeast Asia (SEA). In 2018, the SEA TPE Consortium (SEATPEC) was established, consisting of regional neurologists working to improve outcome of various autoimmune neurological diseases. We proposed an immunotherapeutic guideline prioritizing TPE for this region. We reviewed disease burden, evidence-based treatment options, and major guidelines for common autoimmune neurological disorders seen in SEA. A modified treatment algorithm based on consensus agreement by key-opinion leaders was proposed. Autoimmune antibody diagnostic testing through collaboration with accredited laboratories was established. Choice of first-line immunotherapies (IVIg/corticosteroid/TPE) is based on available evidence, clinicians' experience, contraindications, local availability, and affordability. TPE could be chosen as first-line therapy for GBS, CIDP, MG (acute/short term), IgG, A paraproteinemic neuropathy, and NMDAR encephalitis. Treatment is stopped for acute monophasic conditions such as GBS and ADEM following satisfactory outcome. For chronic immune disorders, a therapy taper or long-term maintenance therapy is recommended depending on the defined clinical state. TPE as second-line treatment is indicated for IVIg or corticosteroids refractory cases of ADEM, NMOSD (acute), MG, and NMDAR/LGI1/CASPR2/Hashimoto's encephalitis. With better diagnosis, treatment initiation with TPE is a sustainable and effective immunotherapy for autoimmune neurological diseases in SEA.
  13. Zangrillo A, Lomivorotov VV, Pasyuga VV, Belletti A, Gazivoda G, Monaco F, et al.
    PMID: 35168907 DOI: 10.1053/j.jvca.2022.01.001
    OBJECTIVE: To investigate the effect of volatile anesthetics on the rates of postoperative myocardial infarction (MI) and cardiac death after coronary artery bypass graft (CABG).

    DESIGN: A post hoc analysis of a randomized trial.

    SETTING: Cardiac surgical operating rooms.

    PARTICIPANTS: Patients undergoing elective, isolated CABG.

    INTERVENTIONS: Patients were randomized to receive a volatile anesthetic (desflurane, isoflurane, or sevoflurane) or total intravenous anesthesia (TIVA). The primary outcome was hemodynamically relevant MI (MI requiring high-dose inotropic support or prolonged intensive care unit stay) occurring within 48 hours from surgery. The secondary outcome was 1-year death due to cardiac causes.

    MEASUREMENTS AND MAIN RESULTS: A total of 5,400 patients were enrolled between April 2014 and September 2017 (2,709 patients randomized to the volatile anesthetics group and 2,691 to TIVA). The mean age was 62 ± 8.4 years, and the median baseline ejection fraction was 57% (50-67), without differences between the 2 groups. Patients in the volatile group had a lower incidence of MI with hemodynamic complications both in the per-protocol (14 of 2,530 [0.6%] v 27 of 2,501 [1.1%] in the TIVA group; p = 0.038) and as-treated analyses (16 of 2,708 [0.6%] v 29 of 2,617 [1.1%] in the TIVA group; p = 0.039), but not in the intention-to-treat analysis (17 of 2,663 [0.6%] v 28 of 2,667 [1.0%] in the TIVA group; p = 0.10). Overall, deaths due to cardiac causes were lower in the volatile group (23 of 2,685 [0.9%] v 40 of 2,668 [1.5%] than in the TIVA group; p = 0.03).

    CONCLUSIONS: An anesthetic regimen, including volatile agents, may be associated with a lower rate of postoperative MI with hemodynamic complication in patients undergoing CABG. Furthermore, it may reduce long-term cardiac mortality.

  14. Shepherdson JL, Hutchison K, Don DW, McGillivray G, Choi TI, Allan CA, et al.
    Am J Hum Genet, 2024 Mar 07;111(3):487-508.
    PMID: 38325380 DOI: 10.1016/j.ajhg.2024.01.007
    Pathogenic variants in multiple genes on the X chromosome have been implicated in syndromic and non-syndromic intellectual disability disorders. ZFX on Xp22.11 encodes a transcription factor that has been linked to diverse processes including oncogenesis and development, but germline variants have not been characterized in association with disease. Here, we present clinical and molecular characterization of 18 individuals with germline ZFX variants. Exome or genome sequencing revealed 11 variants in 18 subjects (14 males and 4 females) from 16 unrelated families. Four missense variants were identified in 11 subjects, with seven truncation variants in the remaining individuals. Clinical findings included developmental delay/intellectual disability, behavioral abnormalities, hypotonia, and congenital anomalies. Overlapping and recurrent facial features were identified in all subjects, including thickening and medial broadening of eyebrows, variations in the shape of the face, external eye abnormalities, smooth and/or long philtrum, and ear abnormalities. Hyperparathyroidism was found in four families with missense variants, and enrichment of different tumor types was observed. In molecular studies, DNA-binding domain variants elicited differential expression of a small set of target genes relative to wild-type ZFX in cultured cells, suggesting a gain or loss of transcriptional activity. Additionally, a zebrafish model of ZFX loss displayed an altered behavioral phenotype, providing additional evidence for the functional significance of ZFX. Our clinical and experimental data support that variants in ZFX are associated with an X-linked intellectual disability syndrome characterized by a recurrent facial gestalt, neurocognitive and behavioral abnormalities, and an increased risk for congenital anomalies and hyperparathyroidism.
  15. Viswanathan S, Appiwatanakul M, Nayak A, Islam B, Khatri B, Pangeran D, et al.
    Ther Apher Dial, 2019 Jun;23(3):289-297.
    PMID: 30927331 DOI: 10.1111/1744-9987.12806
    In conjunction with the third regional Southeast Asian (SEA) therapeutic plasma exchange (TPE) conference in Kuala Lumpur, Malaysia, 25 clinicians and researchers from SEA and South Asian countries attended the inaugural strategy meeting for the establishment of a regional TPE consortium for neurological disorders. The primary objective was to establish regional collaboration to improve delivery of TPE services in SEA. A pre-meeting survey was conducted to gather insights on disease spectrum, contextual practice challenges, and the need for a regional TPE consensus. Challenges identified include limited healthcare funding in support of diagnostic workup, TPE therapy, as well as development of clinical infrastructure and expertise capacity building. There was favorable interest in developing a working plan contextualized to this region. Strategies to overcome challenges were discussed. This included the need for a comprehensive referral system and network of regional TPE centers suited to local needs, supported by innovative TPE delivery programs.
  16. Chazan G, Jupp J, Bauters T, Duncan N, Weddle KJ, Nomura H, et al.
    J Oncol Pharm Pract, 2021 Oct 23.
    PMID: 34693814 DOI: 10.1177/10781552211048892
    INTRODUCTION: The coronavirus of 2019 pandemic has necessitated vast and rapid changes in the way oncology pharmacy services are delivered around the world.

    METHODS/AIMS: An international survey of oncology pharmacists and technicians was conducted via the International Society of Oncology Pharmacy Practitioners and collaborating global pharmacy organisations to determine the impact that the coronavirus of 2019 has had on pharmacy service delivery, pharmacy practitioners and oncology practice.

    RESULTS: The survey received 862 responses from 40 different countries from September to October 2020. The majority of respondents were pharmacists (n = 841, 97.6%), with 24% involved in the direct care of patients with the coronavirus of 2019. Of the survey participants, 55% increased their time working remotely, with remote activities including dispensing, patient assessment/follow-up and attending multi-disciplinary rounds. Respondents reported a 72% increase in the use of technology to perform remote patient interaction activities and that participation in educational meetings and quality improvement projects was reduced by 68% and 44%, respectively. Workforce impacts included altered working hours (50%), cancelled leave (48%) and forced leave/furloughing (30%). During the pandemic, respondents reported reduced access to intensive care (19%) and anti-cancer (15%) medications. In addition, 39% of respondents reported reduced access to personal protective equipment, including N95 masks for chemotherapy compounding. Almost half of respondents (49%) reported that cancer treatments were delayed or intervals were altered for patients being treated with curative intent. A third of practitioners (30%) believed that patient outcomes would be adversely impacted by changes to pharmacy services. Sixty-five percent of respondents reported impacts on their mental health, with 12% utilising support services.

    CONCLUSION: The coronavirus of 2019 pandemic has altered the way oncology pharmacy services are delivered. These results demonstrate the adaptability of the oncology pharmacy profession and highlight the importance of formal evaluation of the varied practice models to determine the evidence-based practices that enhance pharmacy services and, thus, should be reinstated as soon as practical and reasonable.

  17. Zuntini AR, Carruthers T, Maurin O, Bailey PC, Leempoel K, Brewer GE, et al.
    Nature, 2024 Apr 24.
    PMID: 38658746 DOI: 10.1038/s41586-024-07324-0
    Angiosperms are the cornerstone of most terrestrial ecosystems and human livelihoods1,2. A robust understanding of angiosperm evolution is required to explain their rise to ecological dominance. So far, the angiosperm tree of life has been determined primarily by means of analyses of the plastid genome3,4. Many studies have drawn on this foundational work, such as classification and first insights into angiosperm diversification since their Mesozoic origins5-7. However, the limited and biased sampling of both taxa and genomes undermines confidence in the tree and its implications. Here, we build the tree of life for almost 8,000 (about 60%) angiosperm genera using a standardized set of 353 nuclear genes8. This 15-fold increase in genus-level sampling relative to comparable nuclear studies9 provides a critical test of earlier results and brings notable change to key groups, especially in rosids, while substantiating many previously predicted relationships. Scaling this tree to time using 200 fossils, we discovered that early angiosperm evolution was characterized by high gene tree conflict and explosive diversification, giving rise to more than 80% of extant angiosperm orders. Steady diversification ensued through the remaining Mesozoic Era until rates resurged in the Cenozoic Era, concurrent with decreasing global temperatures and tightly linked with gene tree conflict. Taken together, our extensive sampling combined with advanced phylogenomic methods shows the deep history and full complexity in the evolution of a megadiverse clade.
  18. Klionsky DJ, Abdel-Aziz AK, Abdelfatah S, Abdellatif M, Abdoli A, Abel S, et al.
    Autophagy, 2021 Jan;17(1):1-382.
    PMID: 33634751 DOI: 10.1080/15548627.2020.1797280
    In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
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