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Diplazium esculentum, Drynaria quercifolia, and Asplenium nidus are among the fern species found in Bukidnon, Philippines which are used as traditional herbal medicines. The HPTLC profile and the anti-inflammatory properties against cyclooxygenase (COX) and 15-lipoxygenase (15-LOX) of the frond ethanolic extracts of D. esculentum, D. quercifolia, and A. nidus were determined. The High-Performance Thin Layer Chromatography (HPTLC) profile was obtained using ethyl acetate: formic acid: water (16:2:2) as mobile phase and Natural Products (NP) as derivatizing reagent. The HPTLC profile of the D. esculentum, D. quercifolia, and A. nidus extracts showed 10 (Rf = 0.02-0.97), 13 (Rf = 0.03-0.90), and 14 (Rf = 0.02-0.99) bands, respectively. The profiles for each fern species may be used as marker for quality evaluation and standardization of herbal formulations containing these plants. For the anti-inflammatory properties, D. esculentum and D. quercifolia extracts which inhibited more than 50% of the COX enzymes showed significantly higher activity than A. nidus and were considered active against COX-2 and COX-1. D. esculentum, however, gave a selectivity ratio (COX-2/COX-1) of 1.03 making its inhibitory activity selective against COX-2. The percent 15-LOX inhibitory activity of D. quercifolia (58.62%) is significantly higher than that of A. nidus (38.70%) but statistically comparable to D. esculentum (51.19%). Among the extracts, D. quercifolia and D. esculentum which inhibited more than 50% of the 15-LOX were considered active. D. esculentum and D. quercifolia can therefore be potential sources of anti-inflammatory lead compounds for future drug development.

Abstract: With 1.5 million new infections and 690,000 AIDS-related deaths globally each year, HIV- 1 remains a pathogen of significant public health concern. Although a wide array of effective antiretroviral drugs have been discovered, these largely target intracellular stages of the viral infectious cycle, and inhibitors that act at or before the point of viral entry still require further advancement. A unique class of HIV-1 entry inhibitors, called peptide triazoles (PTs), has been developed, which irreversibly inactivates Env trimers by exploiting the protein structure’s innate metastable nature. PTs, and a related group of inhibitors called peptide triazole thiols (PTTs), are peptide compounds that dually engage the CD4 receptor and coreceptor binding sites of Env’s gp120 subunit. This triggers dramatic conformational rearrangements of Env, including the shedding of gp120 (PTs and PTTs) and lytic transformation of the gp41 subunit to a post-fusion-like arrangement (PTTs). Due to the nature of their dual receptor site engagement, PT/PTT-induced conformational changes may elucidate mechanisms behind the native fusion program of Env trimers following receptor and coreceptor engagement, including the role of thiols in fusion. In addition to inactivating Env, PTT-induced structural transformation enhances the exposure of important and conserved neutralizable regions of gp41, such as the membrane proximal external region (MPER). PTT-transformed Env could present an intriguing potential vaccine immunogen prototype. In this review, we discuss the origins of the PT class of peptide inhibitors, our current understanding of PT/PTT-induced structural perturbations and viral inhibition, and prospects for using these antagonists for investigating Env structural mechanisms and for vaccine development.

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Dear EditorThe COVID-19 pandemic had a significant impact on the delivery of medical education across the UK, with the majority of face-to-face teaching cancelled during the first wave, and medical...

A strategy has been established for the synthesis of a family of bifunctional HIV-1 inhibitor covalent conjugates with the potential to bind simultaneously to both the gp120 and gp41 subunits of the HIV-1 envelope glycoprotein trimeric complex (Env). One component of the conjugates is derived from BNM-III-170, a small-molecule CD4 mimic that binds to gp120. The second component, comprised of the peptide DKWASLWNW ("Trp3"), was derived from the N-terminus of the HIV-1 gp41 Membrane Proximal External Region (MPER) and found previously to bind to the gp41 subunit of Env. The resulting bifunctional conjugates were shown to inhibit virus cell infection with low micromolar potency and to induce lysis of the HIV-1 virion. Crucially, virolysis was found to be dependent on the covalent linkage of the BNM-III-170 and Trp3 domains, as coadministration of a mixture of the un-cross-linked components proved to be nonlytic. However, a significant magnitude of lytic activity was observed in Env-negative and other control pseudoviruses, suggesting parallel mechanisms of action of the conjugates involving Env interaction and direct membrane disruption. Computational modeling suggested strong membrane-binding activity of BNM-III-170, which may underly the nonspecific virolytic effects of the conjugates. To investigate the scope of the membrane effect, cell-based cytotoxicity and membrane permeability assays were performed employing flow cytometry. Here, we observed a dose-dependent and specific cytotoxic effect on HIV-1 Env-expressing cells by the small-molecule bifunctional inhibitor. Most importantly, Env-negative cells were not susceptible to the cytotoxic effect upon exposure to this construct at concentrations where cell-killing effects were observed for Env-positive cells. Computational structural modeling supports a mechanism in which the bifunctional inhibitors bind to the gp120 and gp41 subunits in tandem in open-state Env trimers and induce relative motion of the gp120 subunits consistent with models of Env inactivation. This observation supports the idea that the cell-killing effect of the small-molecule bifunctional inhibitor is due to specific Env conformational triggering. This work lays important groundwork to advance a small-molecule bifunctional inhibitor approach for eliminating Env-expressing infected cells and the eradication of HIV-1.

Dual-acting virucidal entry inhibitors (DAVEIs) have previously been shown to cause irreversible inactivation of HIV-1 Env-presenting pseudovirus by lytic membrane transformation. This study examined whether this transformation could be generalized to include membranes of Env-presenting cells. Flow cytometry was used to analyze HEK293T cells transiently transfected with increasing amounts of DNA encoding JRFL Env, loaded with calcein dye, and treated with serial dilutions of microvirin (Q831K/M83R)-DAVEI. Comparing calcein retention against intact Env expression (via Ab 35O22) on individual cells revealed effects proportional to Env expression. "Low-Env" cells experienced transient poration and calcein leakage, while "high-Env" cells were killed. The cell-killing effect was confirmed with an independent mitochondrial activity-based cell viability assay, showing dose-dependent cytotoxicity in response to DAVEI treatment. Transfection with increasing quantities of Env DNA showed further shifts toward "High-Env" expression and cytotoxicity, further reinforcing the Env dependence of the observed effect. Controls with unlinked DAVEI components showed no effect on calcein leakage or cell viability, confirming a requirement for covalently linked DAVEI compounds to achieve Env transformation. These data demonstrate that the metastability of Env is an intrinsic property of the transmembrane protein complex and can be perturbed to cause membrane disruption in both virus and cell contexts.

There are still lot of controversies whether aberrant left hepatic artery (ALHA) originating from left gastric artery should be ligated or preserved during gastric cancer (GC) surgery. We aimed to investigate this issue. We reviewed ALHA cases who had laparoscopic gastrectomy for gastric cancer at Seoul National University Hospital (SNUH) from 2012 to 2016. Type of ALHA variants using Michel’s classification of hepatic arterial anatomy and diameter of each vessel were evaluated by 2 radiologists. Postoperative hepatic function and surgical outcome were collected until 6 months after surgery. Results showed that if the diameter of ALHA was larger than 1.5 mm, a transient elevation of SGOT and SGPT on postoperative day 2 was observed in the ligated cases. No differences were observed in operation time, amount of blood loss, overall complication rate, hospital stay, and number of lymph nodes retrieved between the ligated and preserved replaced left hepatic artery (RLHA) and accessory left hepatic artery (acLHA) group. In this study, we conclude that ligation of ALHA seems to be safe as none of the patients suffered adverse outcome. A transient rise in postoperative SGOT and SGPT levels were seen after ligating ALHA >1.5 mm in diameter regardless of subtype.

KR13, a peptide triazole thiol previously established to inhibit HIV-1 infection and cause virus lysis, was evaluated by flow cytometry against JRFL Env-presenting cells to characterize induced Env and membrane transformations leading to irreversible inactivation. Transiently transfected HEK293T cells were preloaded with calcein dye, treated with KR13 or its thiol-blocked analogue KR13b, fixed, and stained for gp120 (35O22), MPER (10E8), 6-helix-bundle (NC-1), immunodominant loop (50-69), and fusion peptide (VRC34.01). KR13 induced dose-dependent transformations of Env and membrane characterized by transient poration, MPER exposure, and 6-helix-bundle formation (analogous to native fusion events), but also reduced immunodominant loop and fusion peptide exposure. Using a fusion peptide mutant (V504E), we found that KR13 transformation does not require functional fusion peptide for poration. In contrast, simultaneous treatment with fusion inhibitor T20 alongside KR13 prevented membrane poration and MPER exposure, showing that these events require 6-helix-bundle formation. Based on these results, we formulated a model for PTT-induced Env transformation portraying how, in the absence of CD4/co-receptor signaling, PTT may provide alternate means of perturbing the metastable Env-membrane complex, and inducing fusion-like transformation. In turn, the results show that such transformations are intrinsic to Env and can be diverted for irreversible inactivation of the protein complex.

Background:Psoriasis is a chronic inflammatory disease of the skin with a reported prevalence of 0.09-11.4% of the population (1). 1 in 4 psoriasis patients also have psoriatic arthritis (PsA) (2), with additional joint involvement that can be associated with significant morbidity. Despite its relative commonness, the aetiology of psoriasis is not well understood, and there is no cure for this disease. Additionally, up to 30% of PsA patients with active disease are recalcitrant to treatment. Thus it remains a prerogative to understand the immune mechanisms contributing to the development of the disease in order to inform strategies for novel therapies.Objectives:Our aim was to identify perturbations in local tissue immune networks that could contribute to the pathology of psoriasis and psoriatic arthritis. We hypothesise that psoriasis is driven by a disrupted tissue microenvironment, which then provides cues to a susceptible peripheral immune system to drive pathology. Thus as the first part of our study, we investigated the transcriptional profiles of normal and lesional skin.Methods:Skin punch biopsies were obtained from both lesional and morphologically normal skin of 4 PsA patients with active disease. CD45+ cells were isolated using magnetic enrichment for RNA purification and subsequent RNAseq. Differently expressed genes (DEG) were identified and pathway analysis performed using the integrated Differential Expression and Pathway (iDEP) analysis tool. Gene set enrichment analysis was performed using GSEA.Results:Transcriptomic analyses of skin revealed that lesional skin, compared to non-lesional sites, was enhanced for expression of genes associated with immune processes (including genes such as such asIL17A,FCN1, andCTLA4) anti-microbial responses (such asDEF4BAandS100A8) and immune cell chemotaxis (notablyCXCL13andSELPLG), suggesting a possible inflammatory response to skin microbiota. Interestingly, lesional skin showed a deficiency in expression of genes associated with tRNA metabolic processes (includingAARS,YARS, and other aminoacyl tRNA synthetases), suggesting a possible defect in protein translation. Similarly, pathway analysis revealed an enrichment in humoral immune response pathways in PsA lesional skin, and a comparative deficiency in RNA metabolic pathways.Conclusion:Our transcriptional approach provides a comprehensive overview of localised immunity in psoriasis and predicts intimate interactions with the peripheral immune system. Further studies are ongoing to uncover cell types involved, as well as parallels at other disease sites (joints). These findings will facilitate the identification of novel targets for treatment of PsA.References:[1]World Health Organization (2016).Global report on psoriasis. [Online] (https://apps.who.int/iris/bitstream/handle/10665/204417/9789241565189_eng.pdf.psoriasis;jsessionid=54912784D28C9F36ECCD45471AC5775B?sequence=1,accessed 24 January 2020)[2]Alinaghi F., Calov M., Kristensen L.E., Gladman D.D, Coates L.C., Julien D., Gottlieb A.B., Gisondi P., Wu J.J., Thyssen J.P., Egeberg A. (2019) Prevalence of psoriatic arthritis in patients with psoriasis: A systemic review and meta-analysis of observational and clinical studies.Journal of the American Academy of Dermatology.,80(1), 251-265.Disclosure of Interests:Gladys Ang: None declared, Pavanish Kumar: None declared, Dianyan Guo: None declared, Ahmad Lajam: None declared, Warren Fong Consultant of: Abbvie, Janssen, Novartis, Speakers bureau: Abbvie, Janssen, Novartis, Ying Ying Leung Speakers bureau: Novartis, Janssen, Eli Lilly, Salvatore Albani: None declared

OBJECT Long-term memory deficits occur after mild traumatic brain injuries (mTBIs), and effective treatment modalities are currently unavailable. Cerebrolysin, a peptide preparation mimicking the action of neurotrophic factors, has beneficial effects on neurodegenerative diseases and brain injuries. The present study investigated the long-term effects of Cerebrolysin treatment on cognitive function in rats after mTBI. METHODS Rats subjected to closed-head mTBI were treated with saline (n = 11) or Cerebrolysin (2.5 ml/kg, n = 11) starting 24 hours after injury and then daily for 28 days. Sham animals underwent surgery without injury (n = 8). To evaluate cognitive function, the modified Morris water maze (MWM) test and a social odor–based novelty recognition task were performed after mTBI. All rats were killed on Day 90 after mTBI, and brain sections were immunostained for histological analyses of amyloid precursor protein (APP), astrogliosis, neuroblasts, and neurogenesis. RESULTS Mild TBI caused long-lasting cognitive memory deficits in the MWM and social odor recognition tests up to 90 days after injury. Compared with saline treatment, Cerebrolysin treatment significantly improved both long-term spatial learning and memory in the MWM test and nonspatial recognition memory in the social odor recognition task up to 90 days after mTBI (p < 0.05). Cerebrolysin significantly increased the number of neuroblasts and promoted neurogenesis in the dentate gyrus, and it reduced APP levels and astrogliosis in the corpus callosum, cortex, dentate gyrus, CA1, and CA3 regions (p < 0.05). CONCLUSIONS These results indicate that Cerebrolysin treatment of mTBI improves long-term cognitive function, and this improvement may be partially related to decreased brain APP accumulation and astrogliosis as well as increased neuroblasts and neurogenesis.

Fructose is a hexose that when ingested in high amount increases the risk of type 2 diabetes and other metabolic diseases. Type 2 diabetes is estimated to be the 7th leading cause of mortality by 2030, according to the World Health Organization (WHO). The number of people with diabetes has risen from 108 million in 1980 to 422 million in 2014. Food security which is defined as the availability, stability, access and utilization of safe foods is a major determinant in the incidence of type 2 diabetes and other metabolic diseases. Cost-effective food products are highly available and usually utilized abundantly. For example, high fructose corn syrup (HFCS) found in numerous food products such as pastries, yogurt and ice cream are used excessively due to its low cost and versatility. Fructose in its pure form or from HFCS supplemented in our diets can be one of the determining factors that cause type 2 diabetes and other metabolic diseases. Fructose, once ingested, can be oxidized, converted to glucose or converted to lactic acid or enter de novo lipogenesis. These metabolic pathways could lead to the development of type 2 diabetes and metabolic diseases such as obesity and metabolic syndrome. Avoiding excessive intake of fructose can reduce the propensity for type 2 diabetes and other metabolic diseases. In this research study, the methodology from human and animal studies were searched in PubMed and Google Scholar databases. The following key words were searched in each paper: fructose, high fructose corn syrup, sugar, sucrose, metabolic syndrome, type 2 diabetes, insulin resistance, blood glucose, blood sugar, triglycerides, lipoproteins, high density lipoprotein, cholesterol, and low-density lipoprotein. Overall, approximately 150 articles and books were gathered. In this review article, it has been concluded that high fructose ingestion increases risk of metabolic diseases such as type 2 diabetes and the metabolic syndrome. The aim of this research study is to elucidate the role of fructose metabolism in type 2 diabetes and other metabolic diseases.

Thymosin β4 (Tβ4) promotes CNS and peripheral nervous system (PNS) plasticity and neurovascular remodeling leading to neurological recovery in a range of neurological diseases. Treatment of neural injury and neurodegenerative disease 24 h or more post-injury and disease onset with Tβ4 enhances angiogenesis, neurogenesis, neurite and axonal outgrowth, and oligodendrogenesis, and thereby, significantly improves functional and behavioral outcomes. We propose that oligodendrogenesis is a common link by which Tβ4 promotes recovery after neural injury and neurodegenerative disease. The ability to target many diverse restorative processes via multiple molecular pathways that drive oligodendrogenesis and neurovascular remodeling may be mediated by the ability of Tβ4 to alter cellular expression of microRNAs (miRNAs). However, further investigations on the essential role of miRNAs in regulating protein expression and the remarkable exosomal intercellular communication network via exosomes will likely provide insight into mechanisms of action and means to amplify the therapeutic effects of Tβ4.

The aim of this study is to categorize splenic artery and vein configurations, and examine their influence on suprapancreatic lymph node (LN) dissection in laparoscopic gastrectomy. Digital Imaging and Communications in Medicine images from 169 advanced cancer patients who underwent laparoscopic gastrectomy with D2 dissection were used to reconstruct perigastric vessels in 3D using a volume rendering program (VP Planning®). Splenic artery and vein configuration were classified depending on the relative position of their lowest part in regard to the pancreas. Number of resected LNs and surgical outcomes were analyzed. The splenic artery was categorized as superficial (36.7%), middle (49.1%), and concealed (14.2%), and the splenic vein was categorized as superior (6.5%), middle (42.0%), and inferior to the pancreas (51.5%). The number of resected LNs around the proximal half of the splenic artery (#11p) and the proportion of the splenic vein located inferiorly to the pancreas were significantly higher in splenic arteries of concealed types. LN metastasis of station #7 was an independent risk factor of LN metastasis in station #11p (p = 0.010). Concealed types showed a tendency towards longer operating times, more blood loss, longer hospital stays, and a higher postoperative morbidity. Concealed types of splenic artery are associated with an increased difficulty in the dissection of LN station #11p around the splenic artery. A 3D volume rendering program is a useful tool to rapidly and intuitively identify individual anatomical variations, to plan a tailored surgical strategy, and to predict potential challenges.

One of the laboratory tests recommended by the American Diabetes Association (ADA) to screen for diabetes mellitus (DM) is HbA1c, and it is particularly suitable for segments of the population that cannot or are unwilling to fast for a screening test. The aim of this study was to determine whether HbA1c would be a useful tool to screen for DM in a real-world setting if ADA guidelines for repeat testing to confirm the diagnosis of DM are strictly adhered to.A retrospective database study was performed by extracting demographic and laboratory data from a chronic disease registry that collects data on adults from three tertiary hospitals and nine large primary care clinics in Singapore. Data were extracted and analyzed for adults not previously known to have DM whose data was captured in the registry between 2005 and 2016 with HbA1c and at least two diagnostic tests for DM (fasting plasma glucose or 2-h plasma glucose) performed within 4 weeks after HbA1c determination.In all, 3928 adults were included in this study. The sensitivity, specificity, and area under the receiver operating characteristic curve for HbA1c at a threshold of 6.5% were 85.2%, 82.3%, and 0.914, respectively. A higher sensitivity was found in female adults, younger adults, and those of non-Chinese ethnicity.The sensitivity of HbA1c as a screening test for DM in this study was significantly higher than that reported previously. This work provides additional evidence supporting the inclusion of HbA1c as one of the screening tests for DM.

The mean annual direct medical cost of type 2 diabetes mellitus (T2DM) in Singapore has been found to be SGD 2034 using the prevalence-based approach, but the lifetime direct medical cost of T2DM in Singapore remains largely unknown. The aim of the present study was to determine the lifetime direct medical cost attributable to T2DM and provide estimates of potential savings if T2DM can be prevented or delayed.The incidence-based approach was used for the cost-of-illness analysis. Yearly medical expenses were obtained from a regional health system database in Singapore to estimate the lifetime medical cost of T2DM patients. Then, the lifetime medical cost of non-T2DM subjects was predicted using a regression model. From the database, gender- and age-specific annual survival rates of T2DM and non-T2DM subjects were obtained and survival-adjusted yearly expenses over the estimated remaining life span were added to obtain lifetime medical costs. The difference between T2DM and non-T2DM subjects was attributed to excess direct medical costs of T2DM.The excess lifetime medical expenses for T2DM patients were SGD 132 506, 108 589, 83 326 and 70 110 when the age of T2DM diagnosis was 40, 50, 60, and 65 years, respectively.Even though T2DM patients have a lower life expectancy, T2DM is associated with substantially higher lifetime medical costs. Delaying the onset of T2DM, especially in the young, may lead to lower lifetime medical expenses. If prevention costs can be kept sufficiently low, effective T2DM prevention efforts would likely lead to a reduction in long-term medical costs.