Tetrahydropalmatine from medicinal plants activates human glucokinase to regulate glucose homeostasis.

Many synthetic glucokinase activators (GKAs), modulating glucokinase (GK), an important therapeutic target in diabetes have failed to clear clinical trials. In this study, an in silico structural similarity search with differing scaffolds of reference GKAs have been used to identify derivatives from natural product databases. Ten molecules with good binding score and similar interactions to that in the co-crystallized GK as well good activation against recombinant human GK experimentally were identified. Tetrahydropalmatine, an alkaloid present in formulations and drugs from medicinal plants, has not been explored as an antidiabetic agent and no information regarding its mechanism of action or GK activation exists. Tetrahydropalmatine activates GK with EC50 value of 71.7 ± 17.9 µM while lowering the S0.5 (7.1 mM) and increasing Vmax (9.22 µM/min) as compared to control without activator (S0.5 = 10.37 mM; Vmax = 4.8 µM/min). Kinetic data (α and ß values) suggests it to act as mixed, nonessential type activator. Using microscale thermophoresis, Kd values of 3.8 µM suggests a good affinity for GK. In HepG2 cell line, the compound potentiated the uptake of glucose and maintained glucose homeostasis by increasing the expression of GK, glycogen synthase, and insulin receptor genes and lowering the expression of glucokinase regulatory protein (GKRP) and glucagon. Tetrahydropalmatine at low concentrations could elicit a good response by reducing expression of GKRP, increasing expression of GK while also activating it. Thus, it could be used alone or in combination as therapeutic drug as it could effectively modulate GK and alter glucose homeostasis.

PRpnp, a novel dual activity PNP family protein improves plant vigour and confers multiple stress tolerance in Citrus aurantifolia.

Under field conditions, plants are often simultaneously exposed to several abiotic and biotic stresses resulting in significant reductions in growth and yield; thus, developing a multi-stress tolerant variety is imperative. Previously, we reported the neofunctionalization of a novel PNP family protein, Putranjiva roxburghii purine nucleoside phosphorylase (PRpnp) to trypsin inhibitor to cater to the needs of plant defence. However, to date, no study has revealed the potential role and mechanism of either member of this protein group in plant defence. Here, we overexpressed PRpnp in Citrus aurantifolia which showed nuclear-cytoplasmic localization, where it functions in maintaining the intracellular purine reservoir. Overexpression of PRpnp significantly enhanced tolerance to salt, oxidative stress, alkaline pH, drought and two pests, Papilio demoleus and Scirtothrips citri in transgenic plants. Global gene expression studies revealed that PRpnp overexpression up-regulated differentially expressed genes (DEGs) related to ABA- and JA-biosynthesis and signalling, plant defence, growth and development. LC-MS/MS analysis validated higher endogenous ABA and JA accumulation in transgenic plants. Taken together, our results suggest that PRpnp functions by enhancing the endogenous ABA and JA, which interact synergistically and it also inhibits trypsin proteases in the insect gut. Also, like other purine salvage genes, PRpnp also regulates CK metabolism and increases the levels of CK-free bases in transgenic Mexican lime. We also suggest that PRpnp can be used as a potential candidate to develop new varieties with improved plant vigour and enhanced multiple stress resistance.

Efficient transformation and regeneration of transgenic plants in commercial cultivars of Citrus aurantifolia and Citrus sinensis.

Citrus is one of the major horticultural crops with high economic and nutraceutical value. Despite the fact that conventional research has developed numerous improved varieties, citriculture is still susceptible to various stresses and requires innovative solutions such as genetic engineering. Among all the currently available modern approaches, Agrobacterium-mediated transformation is the most efficient method for introducing desired traits in citrus. However, being a non-host for Agrobacterium, various citrus species, including Citrus aurantifolia and Citrus sinensis, are recalcitrant to this method. The available reports on Agrobacterium-mediated transformation of commercial citrus cultivars show very low transformation efficiency with poor recovery rates of whole transgenic plantlets. Here, we provide an efficient and reliable procedure of Agrobacterium-mediated transformation for both C. aurantifolia and C. sinensis. This protocol depends on providing callus-inducing treatment to explants before and during Agrobacterium co-cultivation, using optimum conditions for shoot regeneration and modifying in-vitro micrografting protocol to combat the loss of transgenic lines. As transgenic citrus shoots are difficult to root, we also developed the ideal conditions for their rooting. Using this protocol, the whole transgenic plantlets of C. aurantifolia and C. sinensis can be developed in about ~ 4 months, with transformation efficiency of 30% and 22% for the respective species.

Tranexamic acid (class I drug) reduced and capped gold nanoparticles as a potential hemostatic agent with enhanced performance.

External hemostatic agents play a crucial role in stabilizing an impaired process during pathological conditions. The idea is to stabilize thein vivosystem as soon as possible. This study uses a class I hemostatic drug tranexamic acid as a reducing and capping agent for synthesizing the gold nanoparticles (Tr-AuNPs). Being the synthetic analogue of lysine and a biologically inspired alkylamine molecule, the chemistry can be fine-tuned for stable material that can simultaneously target the intrinsic and extrinsic hemostatic pathway, making it promising for hemostatic applications. The Tr-AuNPs of hydrodynamic diameter ∼46 nm were synthesized and evaluated physio-chemically using various analytical techniques wherein they showed hemocompatibility and increased thrombus weight compared to the native drug. The decrease in prothrombin time (PT) and international normalized ratio supported by the dynamic thromboelastography (TEG) study indicates the prepared nano-conjugate's potential in reducing time for attaining hemostasis as compared to the native tranexamic acid drug. At a 9µg ml-1concentration, Tr-AuNPs had a procoagulant effect, shown by decreased reaction time (R) and coagulation time (K) with improvedαangle and MA. There was a significant increase in the rate of coagulationin vivoby Tr-AuNPs, i.e. (52 s) compared to the native tranexamic acid (360 s). Radiolabelling studies ascertained thein vivobiocompatibility (non-invasive distribution, residence, clearance, and stability) of the Tr-AuNPs. The short-term toxicity studies were conducted to establish a proof of concept for the biomedical application of the material. The results highlighted the use of biologically alkyl amine molecules as capping and reducing agents for the synthesis of nanoparticles, which have shown a synergistic effect on the coagulation cascade while holding the potential for also acting as potential theranostic agents.

The Effect of Prophylactic Steroids on Shivering in Adults Undergoing Surgery: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

BACKGROUND: Shivering is common following anesthesia and surgery. Corticosteroids (steroids) have been tried to reduce the risk of shivering, but the evidence in favor of their use is uncertain. The primary objective of this review was to evaluate the effect of steroids on the risk of perioperative (intra- and postoperative) shivering versus controls (placebo and active controls). Additional objectives were to assess the risk of severity of shivering, patient satisfaction with shivering prophylaxis, quality of recovery (QoR), and the risk of steroid-related adverse effects. METHODS: PubMed, Embase, Cochrane Central Registry of Trials, Google Scholar, and preprint servers were searched from inception until November 30, 2022. Randomized controlled trials (RCTs) published in the English language were retrieved, provided they reported on shivering either as a primary or secondary outcome following steroid prophylaxis in adult patients undergoing surgery under spinal or general anesthesia. RESULTS: A total of 3148 patients from 25 RCTs were included in the final analysis. The steroids used in the studies were either dexamethasone or hydrocortisone. Dexamethasone was administered intravenously or intrathecally, while hydrocortisone was administered intravenously. Prophylactic administration of steroids reduced the risk of overall shivering (risk ratio [RR], 0.65 [95% confidence interval {CI}, 0.52-0.82]; P = .0002; I2 = 77%) as well as the risk of moderate to severe shivering (RR, 0.49 [95% CI, 0.34-0.71]; P = .0002; I2 = 61%) in comparison to controls. Administration of intravenous dexamethasone (RR, 0.67 [95% CI, 0.52-0.87]; P = .002; I2 = 78%) and hydrocortisone (RR, 0.51 [95% CI, 0.32-0.80]; P = .003; I2 = 58%) were effective in shivering prophylaxis. For intrathecal dexamethasone (RR, 0.84 [95% CI, 0.34-2.08]; P = .7; I2 = 56%), the null hypothesis of no subgroup difference was not rejected ( P = .47), preventing definitive conclusions about the efficacy of this route of administration. The prediction intervals for both overall shivering risk (0.24-1.70) and risk of severity of shivering (0.23-1.0) precluded generalization of results in future studies. Meta-regression analysis was used to further explore heterogeneity. Factors like the dose and timing of administration of steroids or the type of anesthesia were not found to be significant. Patient satisfaction and QoR were higher in the dexamethasone groups versus placebo. No increased risk of adverse events of steroids was noted versus placebo or controls. CONCLUSIONS: Prophylactic steroid administration may be beneficial in reducing the risk of perioperative shivering. However, the quality of evidence in favor of steroids is very low. Further well-designed studies are needed for establishing generalization.

A Case of Cobra Bite in a Term Pregnant Woman: The Obstetric and Wound Management Challenges.

Snake envenomation is a rare incident during pregnancy and potentially challenging to manage. Snakebites in pregnancy may lead to several complications such as teratogenicity, miscarriage, antepartum hemorrhage, and even intrauterine fetal death. Here, we report a case of a pregnant woman who presented to our emergency department with signs of systemic envenomation following an Indian cobra bite on her foot, highlighting the key obstetric and wound management challenges. She complained of severe pain at the site of the bite and progressive swelling, abdominal pain, and multiple episodes of vomiting, which started 45 min after the bite. She received 10 vials of polyvalent antivenom from a primary hospital and was then referred to our center. The patient underwent emergency cesarean section and later fasciotomy with free-flap reconstruction at the bitten site due to local tissue necrosis. The case was successfully managed by a multidisciplinary team consisting of an emergency physician, obstetrician, and plastic surgeon, saving 2 lives and the limb of the patient.

The burden of paraovarian cysts - a case series and review of the literature.

Introduction: A paraovarian cyst (POC) is a cyst in the broad ligament or mesosalpinx. Paraovarian cysts have an estimated prevalence of 5-20% amongst the adnexal masses. Despite the high prevalence and availability of advanced imaging modalities, an accurate pre-operative diagnosis of POC is still made in less than 50% of patients. Case reports: Two females with suspected ovarian torsion underwent laparotomy and had POCs. A 42-year-old hysterectomized female underwent surgery for a suspected POC which turned out to be a mesenteric cyst. Two females underwent laparotomy for suspected mesenteric cysts which turned out to be POCs. A nulliparous female with infertility conceived spontaneously after cystectomy of POC. Results: Optimal management of an adnexal mass depends on the knowledge of the origin and the exact nature of the mass. No clear-cut guidelines exist for the management of POCs despite their high prevalence. There is a need for further research on this topic to formulate clear-cut guidelines for their management. Conclusions: Radiologists and gynaecologists need to keep them in mind as differentials for patients with adnexal masses to ensure a correct pre-operative diagnosis in order to achieve an optimal outcome for these females. Gynaecologists need to be aware of the cases which can be managed conservatively and those that need surgery, along with the extent of the surgery required, taking care to protect the ovary at all costs, particularly in benign cases.

Meconium Peritonitis Presenting with Nonimmune Hydrops and Mirror Syndrome.

Meconium peritonitis (MP) presenting as hydrops is a rare entity. A 34-week hydropic infant was born to mother diagnosed with a case of nonimmune hydrops. Postnatally, the neonate was diagnosed as a case of MP based on clinical examination and investigations. The neonate underwent exploratory laparotomy which revealed diffuse MP with ileal perforation. She underwent resection of ileal perforation with ileocolic anastomosis. The index case highlights the importance of thorough clinical examination and abdominal X-ray in the diagnosis of MP.

A brief molecular insight of COVID-19: epidemiology, clinical manifestation, molecular mechanism, cellular tropism and immuno-pathogenesis.

In December 2019, the emergence and expansion of novel and infectious respiratory virus SARS-CoV-2 originated from Wuhan, China caused an unprecedented threat to the public health and became a global pandemic. SARS-CoV-2 is an enveloped, positive sense and single stranded RNA virus belonging to genera betacoronavirus, of Coronaviridae family. The viral genome sequencing studies revealed 75-80% similarity with SARS-CoV. SARS-CoV-2 mainly affects the lower respiratory system and may progress to pneumonia and Acute Respiratory Distress Syndrome (ARDS). Apart from life-threatening situations and burden on the global healthcare system, the COVID-19 pandemic has imposed several challenges on the worldwide economics and livelihood. The novel pathogen is highly virulent, rapidly mutating and has a tendency to cross the species boundaries such as from bats to humans through the evolution and natural selection from intermediate host. In this review we tried to summarize the overall picture of SARS-CoV-2 including origin/ emergence, epidemiology, pathogenesis, genome organization, comparative analysis with other CoVs, infection and replication mechanism along with cellular tropism and immunopathogenesis which will provide a brief panoramic view about the virus and disease.

Synthesis and biological evaluation of modified laminin peptide (N2S2-KDP) with enhanced affinity for neuronal growth and targeted molecular imaging (SPECT).

An analog of γ1 laminin (RDIAEIIKDI) decapeptide has been used to augment neuronal survival and regeneration after injuries, during aging and other CNS disorder. As a prime synthetic peptide, KDI, is responsible for the neurite outgrowth of human embryonic neurons. In this study, we have designed, modified a KDI derivative and synthesized by replacing isoleucine (I) with Pro (P) amino acid at C-terminal to enhance its potency towards neurite growth. -Cys-Gly-Cys (-CGC) N2S2 motif was also incorporated in the present design for peptide radiolabeling. The modified peptide showed a better binding with the desired 3T1M receptor for neurite growth. The peptide was synthesized using solid phase peptide synthesis and Fmoc-strategy with more than 80% yield. The receptor binding studies of 99mTc-N2S2-KDP in Neuro2A cell lines showed Kd value in 31 nM range and the complex showed appreciable brain uptake in mice. The results on human SH-SY5Y indicate that the unlabeled N2S2-KDP may perhaps be useful for neurite growth in neurodegenerative disorder.

The Second- vs First-wave COVID-19: More of the Same or a Lot Worse? A Comparison of Mortality between the Two Waves in Patients Admitted to Intensive Care Units in Nine Hospitals in Western Maharashtra.

BACKGROUND: India, along with the rest of the world, faced the challenging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. The second wave in India lagged behind that in the Western world, due to different timing of seasons. There is scarce data about the differences between the two waves, for intensive care unit (ICU) patients. We present the data of 3,498 patients from 9 ICUs of western Maharashtra. MATERIALS AND METHODS: We collected prospective data of hospitalized, RT-PCR confirmed, coronavirus-2019 (COVID-19) patients, from nine tertiary centers, after institutional ethics committee (IEC) approval. Then, we segregated and analyzed the data of patients admitted to the ICU, for comorbidities, high-resolution computed tomography (HRCT) score, ventilatory support, etc. The primary outcomes were ICU and hospital mortality. We also performed multivariable analysis for predictors of ICU mortality. RESULTS: Overall, there were 3,498 ICU patients. In the first wave, 1,921 patients needed ICU admission, while in the second wave, 1,577 patients. Patients in the second wave had significantly higher ICU (26.1 vs 13.4%, p <0.001) and hospital mortality (29.9 vs 18.2%, p <0.001) and need for ventilatory support of any type. More patients received steroids during the second wave. On multivariable regression, male gender, ICU admission during the second wave, increasing HRCT score, and need for intubation and mechanical ventilation were significant predictors of ICU mortality. CONCLUSION: ICU patients admitted during the two waves were of the similar age, but there were more females, and more patients had comorbidities during the second wave. The ICU and hospital mortality were significantly higher during the second wave. HOW TO CITE THIS ARTICLE: Zirpe KG, Dixit S, Kulkarni AP, Pandit RA, Ranganathan P, Prasad S, et al. The Second- vs First-wave COVID-19: More of the Same or a Lot Worse? A Comparison of Mortality between the Two Waves in Patients Admitted to Intensive Care Units in Nine Hospitals in Western Maharashtra. Indian J Crit Care Med 2021; 25(12):1343-1348.

Tryptophan conjugated magnetic nanoparticles for targeting tumors overexpressing indoleamine 2,3 dioxygenase (IDO) and L-type amino acid transporter.

Tryptophan is an amino acid required by all life forms for protein synthesis and other important metabolic functions. It is metabolized in the body using the kynurenine pathway which involves the enzyme indoleamine 2,3 dioxygenase (IDO) and its transport is regulated through the L-type amino acid transporters (LAT 1). IDO and LAT 1 are found to be overexpressed in many cancers i.e., ovarian, lung colorectal etc. In this study we have used this specific interaction as the basis for designing diagnostic agent based on iron oxide nanoparticles which can specifically target the IDO/LAT 1 over expressing tumors. We have conjugated tryptophan to the surface of super-paramagnetic nanoparticles chemically using 3-aminopropyltrimethoxysilane as a linker. The synthesized tryptophan conjugated magnetic nano-conjugate has been characterized using FTIR, UV-Vis, TEM for its shape size, charge and NMR and Mass for conjugation. The magnetization studies show decrease in the magnetic behavior after conjugation however the desired super-paramagnetic property is still retained as shown by the signature sigmoidal B-H curve. The nano-conjugate shows minimal cytotoxicity over 24 h as shown by the SRB assay in two cell lines A-549, MCF-7. Using 99mTc labeling the biodistribution and the blood kinetics of the magnetic nano-conjugate was evaluated. The study highlights the suitability of the designed magnetic Nano bioconjugate as a potential bimodal diagnostic agent.

A Comparative In Vitro Sensitivity Study of "Ceftriaxone-Sulbactam-EDTA" and Various Antibiotics against Gram-negative Bacterial Isolates from Intensive Care Unit.

INTRODUCTION: A rapid increase in multidrug-resistant (MDR) strains is being seen across the globe especially in the Southeast Asian region, including India. Carbapenems and colistin form the mainstay of treatment against gram-negative pathogens, especially extended-spectrum beta-lactamase (ESBL)- and metallo-beta-lactamse (MBL)-producing isolates. However, due to increased resistance to carbapenems and toxicity of colistin, especially in intensive care units (ICUs), carbapenem-sparing antibiotics like ceftriaxone-sulbactam-EDTA (CSE) combination needs to be evaluated. MATERIALS AND METHODS: Bacterial isolates cultured from various clinical samples from all ICUs for a period of 9 months were evaluated. Bacterial identification was performed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and antibiotic susceptibility testing were performed by disk diffusion and E test method. Antibiogram of various antibiotics was noted. Extended-spectrum beta-lactamase- and MBL-producing bacteria were identified by phenotypic methods. Antibiotic sensitivity results of CSE were compared with the comparator drugs like colistin, carbapenems, and tigecycline in Enterobacteriaceae, Acinetobacter spp., and Pseudomonas spp. along with ESBL and MBL producers. RESULTS: A total of 2,760 samples of blood, cerebrospinal fluid (CSF), respiratory samples, tissue, and pus were collected from ICUs with maximum isolates from pus (37%) followed by respiratory samples (31%) and blood (27%). Escherichia coli and Klebsiella pneumoniae were the predominant gram-negative pathogens accounting for 56% of the isolates followed by Acinetobacter spp. (23%) and Pseudomonas spp. (15%). Extended-spectrum beta-lactamase screening was positive for 57% (1,069/1,877) isolates; whereas 43% (732/1,877) were MBL producers. According to the antibiotic susceptibility results, CSE was the most effective antibiotic showing 94% sensitivity for carbapenem-sensitive Enterobacteriaceae and 97% for carbapenem-resistant Acinetobacter and Pseudomonas spp. Among the other drugs, colistin was found to be the most effective showing almost 95% sensitivity in both the Enterobacteriaceae and non-Enterobacteriaceae group (both ESBL + OXA/NDM). Ceftriaxone-sulbactam-EDTA was also found much more effective (95%) as compared to Colistin (89%) toward ESBL- and MBL-producing strains of Enterobacteriaceae and non-Enterobacteriaceae group. Among the carbapenems, imipenem was the most effective drug against Enterobacteriaceae showing 34% sensitivity and ertapenem proved to be least effective. CONCLUSION: In our present study, CSE emerged as a potent antibacterial agent against MDR gram-negative infections; both for ESBL as well as MBL producers. Hence, in light of present study, we strongly recommend inclusion of CSE in routine sensitivity panel and may be used as a carbapenem- and colistin-sparing drug and a promising option against ESBL and MBL producers especially in ICU. HOW TO CITE THIS ARTICLE: Singh S, Sahu C, Patel SS, Singh A, Yaduvanshi N. A Comparative In Vitro Sensitivity Study of "Ceftriaxone-Sulbactam-EDTA" and Various Antibiotics against Gram-negative Bacterial Isolates from Intensive Care Unit. Indian J Crit Care Med 2020;24(12):1213-1217.

Quadrantic vortex vein decompression with subretinal fluid drainage for manangement of Nanophthalmic choroidal effusions- a review of literature and case series.

BACKGROUND: Uveal effusion syndrome is a rare entity of idiopathic exudative detachments of uveal tissues and retina. Medical treatments with systemic steroids and antimetabolites have been tried but with variable results. Scleral windows or vortex decompressions are usually performed and surgeons usually perform partial sclerectomy in all the quadrants. CASE PRESENTATION: For the first time, we report 2 cases of nanophthalmic uveal effusion syndrome managed with our technique. CONCLUSION: Quadrantic vortex vein decompression with external drainage for nanophthalmic uveal effusion can provide immediate and stable gain in vision.

Riga-Fede Disease Associated with Syndactyly and Oligodactyly: A rare Occurrence.

Background: Eruption of first primary tooth starts on an average at around six to seven months of age. Presence of teeth at birth or within a month after birth is considered rare. Natal teeth are those present in the oral cavity at the time of birth. These teeth can cause ulcers on the ventral surface of the tongue, lip, and the mother's breast characterizing the Riga Fede Disease. Exact aetiology of natal and neonatal teeth is still unclear. The treatment depends on its mobility of teeth as it is associated with the risk of aspiration or swallowing, whether the natal tooth is supernumerary or primary, causing any problems in breast feeding, presence of soft tissue injuries on tongue of the child or mother's breast and overall health of child. Case presentation: A fifteen day old girl reported with large ulceration on ventral surface of tongue due to sharp natal teeth present at mandibular anterior region which was also associated with feeding difficulties. Along with Riga Fede disease Syndactyly and Oligodactyly in left and right legs respectively was also observed. Extraction of the teeth has been carried out and complete healing of ulceration has been achieved within 30 days. Conclusion: The present case report describes a rare occurrence of Riga Fede disease with Syndactyly and Oligodactyly and highlights its symptomatology and therapeutic approach.

Awareness of legal and social issues related to reproductive health among adolescent girls in rural Varanasi.

BACKGROUND: Data on awareness of adolescent's on the legal and social issues/acts related to reproductive health, especially in rural areas, are scarce. OBJECTIVES: The aim of the present cross-sectional study is to assess the awareness level of legal and social issues related to reproductive health and its association with the various individual and family/household level characteristics. METHODS: 650 adolescent girls in the age group of 15-19 years were interviewed with the help of pretested and semistructured questionnaire and focus group discussions were also conducted for qualitative findings in Chiraigaon block of district Varanasi. RESULTS: It was observed that 42.9% of the respondents were aware of legal age of marriage, 14.9% knew about the right age of childbearing. Dowry prohibition act and domestic violence act were known to 46% and 27% respondents, respectively, and only 2.6% were aware of medical termination of pregnancy act. Logistic regression analysis shows the significant effect of education on awareness of legal age of marriage, right age of childbearing, domestic violence, and dowry prohibition acts, which is also supported by qualitative findings. CONCLUSION: All the important legal issues/acts should be included in high school curriculum and female teachers should be involved in training program for adolescents. Role of mass media in creating awareness about these issues in their routine programs should be ascertained. Accredited Social Health Activist and Anganwadi workers should be aware of and include these issues/acts in adolescent meetings.

A comparative study to correlate between clinically and radiographically determined sagittal condylar guidance in participants with different skeletal relationships.

CONTEXT: Many authors have conducted studies that determine horizontal condylar guidance (HCG) using various methods, articulator systems, and recording materials. However, there is a dearth of literature on variability existing in HCG in individuals with different skeletal relationships. This study is an attempt to verify whether such a difference exists or not. AIMS: The aim of this study is to determine and correlate the HCG in individuals with Angle's Class I, Class II, and Class III malocclusion using radiographic and clinical methods. SETTINGS AND DESIGN: HCG was recorded for thirty individuals, ten of each class. For each individual, HCG was recorded clinically as well as radiographically. SUBJECTS AND METHODS: Clinically, HCG was recorded using protrusive check bites and a semi-adjustable articulator. Radiographically, two methods were employed. First, a "tangent method" wherein the angle made by a tangent to the posterior slope of articular eminence with the Frankfurt horizontal (FH) plane was considered as the HCG, and second, a "protrusive method" where the position of the condyle at maximum intercuspation and 6 mm protrusion were traced, and the angle this path made with the FH plane was recorded as the HCG. STATISTICAL ANALYSIS: Descriptive statistical analysis along with Tukey's test and analysis of variance was used to calculate and compare the mean values. Pearson correlation coefficient was used to establish correlation between various means. RESULTS: A significant difference in the HCG of three skeletal relationships was seen, with Class II having a steeper angle than the other two. Among the various methods used, a correlation was found between the clinical and the protrusive method; however, the tangent method yielded greater values of HCG. CONCLUSIONS: The average value of HCG should not be used as it differs according to the skeletal relationship. Radiographic method can be used to yield consistent HCG; however, the protrusive method should be employed.