Marker-free characterization of full-length transcriptomes of single live circulating tumor cells.

The identification and characterization of circulating tumor cells (CTCs) are important for gaining insights into the biology of metastatic cancers, monitoring disease progression, and medical management of the disease. The limiting factor in the enrichment of purified CTC populations is their sparse availability, heterogeneity, and altered phenotypes relative to the primary tumor. Intensive research both at the technical and molecular fronts led to the development of assays that ease CTC detection and identification from peripheral blood. Most CTC detection methods based on single-cell RNA sequencing (scRNA-seq) use a mix of size selection, marker-based white blood cell (WBC) depletion, and antibodies targeting tumor-associated antigens. However, the majority of these methods either miss out on atypical CTCs or suffer from WBC contamination. We present unCTC, an R package for unbiased identification and characterization of CTCs from single-cell transcriptomic data. unCTC features many standard and novel computational and statistical modules for various analyses. These include a novel method of scRNA-seq clustering, named deep dictionary learning using k-means clustering cost (DDLK), expression-based copy number variation (CNV) inference, and combinatorial, marker-based verification of the malignant phenotypes. DDLK enables robust segregation of CTCs and WBCs in the pathway space, as opposed to the gene expression space. We validated the utility of unCTC on scRNA-seq profiles of breast CTCs from six patients, captured and profiled using an integrated ClearCell FX and Polaris workflow that works by the principles of size-based separation of CTCs and marker-based WBC depletion.

Clinical significance of SPOP and APC gene alterations in colorectal cancer in Indian population.

Speckle-Type Poz Protein (SPOP) involved in the regulation of proteasome-mediated degradation of several oncoproteins, resulting in cancer initiation and progression. Mutations in Adenomatous Polyposis Coli (APC) gene is reported in most sporadic and hereditary colorectal cancer (CRC). Identifying the cellular changes involved in carcinogenesis when APC is mutated is an important issue that needs attention. The tumor suppressive function of SPOP and APC has long been a major focus in the research field of colorectal cancer. However, the clinical significance of SPOP and APC gene alteration in CRC has not been established to date. Mutational analysis was performed by single-strand conformational polymorphism followed by Sanger sequencing, methylation status by methylation-specific PCR, and protein expression by immunohistochemistry on 142 tumor tissues along with their adjacent non-cancerous specimens. The overall survival (OS) and recurrence free survival (RFS) were estimated by Kaplan-Meier Curve. Mutation rates of APC and SPOP gene were 2.8% and 11.9% while that of promoter hypermethylation were 37% and 47%, respectively. The grade of differentiation and Lymph node metastasis were significantly correlated with APC methylation pattern (p ≤ 0.05). The down regulation of APC was more often seen in colonic cancer compared to rectal cancer (p = 0.07) and more commonly in T3-4 depth of invasion (p = 0.07) and in patients without lymphovascular and perineural invasion (p = 0.007, p = 0.08 respectively). The median overall survival and recurrence free survival (RFS) was 67 & 36 months while 3-yr and 5-yr OS and RFS were 61.1% & 56.4% and 49.2% & 44.8%, respectively. APC promoter methylation had a better overall survival (p = 0.035) while loss of SPOP expression had a worse survival (p = 0.09). Our findings reveal high percentage of SPOP gene mutations in CRC. A significant link is found between promoter hyper methylation and protein expression in all mutant cases of APC and SPOP, suggesting that both genes may be associated in the development of colorectal cancer in people of Indian decent. Hypermethylation of APC gene and loss of SPOP expression have shown an association with disease prognosis and could be further studied looking at its potential role in planning adjuvant treatment in CRC patients.

Mannose-binding lectin gene 2 variant DD (rs 5030737) is associated with susceptibility to COVID-19 infection in the urban population of Patna City (India).

The study aimed to measure plasma levels of Mannose-Binding Lectin (MBL) and MBL-associated serine protease-2 (MASP-2) and their polymorphisms in COVID-19 patients and controls to detect association. As MBL is a protein of immunological importance, it may contribute to the first-line host defence against SARS-CoV-2. MBL initiates the lectin pathway of complement activation with help of MASP-1 and MASP-2. Hence, appropriate serum levels of MBL and MASPs are crucial in getting protection from the disease. The polymorphisms of MBL and MASP genes affect their plasma levels, impacting their protective function and thus may manifest susceptibility, extreme variability in the clinical symptoms and progression of COVID-19 disease. The present study was conducted to find plasma levels and genetic variations in MBL and MASP-2 in COVID-19 patients and controls using PCR-RFLP and ELISA, respectively.The present study was conducted to find plasma levels and genetic variations in MBL and MASP-2 in COVID-19 patients and controls using PCR-RFLP and ELISA, respectively. Our results indicate that median serum levels of MBL and MASP-2 were significantly low in diseased cases but attained normal levels on recovery. Only genotype DD was found to be associated with COVID-19 cases in the urban population of Patna city.

Zingiber officinale Roscoe Essential Oils-Loaded Chitosan Nanoparticles with Enhanced Bactericidal Efficacy against Burkholderia glumae in Rice.

Ginger essential oils (GEO) shows exceptional antimicrobial properties against plant pathogens. Due to its high volatility and low stability, it requires encapsulation to retain its effective properties. The GEO-Chitosan (GEO-CS) nanobactericide was developed using the ionic gelation method. The nanobactericides show particle diameters of 465, 28, 35, 48 and 500 nm when sodium tripolyphosphate (TPP) concentrations used in the preparation were 0.0, 0.5, 1.0, 2.0 and 4.0 %, respectively. The X-ray diffraction and the UV-vis studies revealed that the GEO was encapsulated into the chitosan nanoparticles with an encapsulation efficiency of around 46 % and a loading capacity of 27-34 %. The antibacterial activity of GEO-chitosan nanobactericide against Burkholderia glumae (Bg) was found to be 7.5-11.8 mm, with minimum inhibitory concentration and minimum bactericidal concentration values of 15.6 µl/mL and 31.25 µl/mL, respectively. Hence, these findings indicate that the prepared GEO-CS nanobactericides were found to be effective against Bg. This preliminary study is toward the development of new agronanobactericides using a natural product to control Bg.

A Sensorised Glove to Detect Scratching for Patients with Atopic Dermatitis.

In this work, a lightweight compliant glove that detects scratching using data from microtubular stretchable sensors on each finger and an inertial measurement unit (IMU) on the palm through a machine learning model is presented: the SensorIsed Glove for Monitoring Atopic Dermatitis (SIGMA). SIGMA provides the user and clinicians with a quantifiable way of assaying scratch as a proxy to itch. With the quantitative information detailing scratching frequency and duration, the clinicians would be able to better classify the severity of itch and scratching caused by atopic dermatitis (AD) more objectively to optimise treatment for the patients, as opposed to the current subjective methods of assessments that are currently in use in hospitals and research settings. The validation data demonstrated an accuracy of 83% of the scratch prediction algorithm, while a separate 30 min validation trial had an accuracy of 99% in a controlled environment. In a pilot study with children (n = 6), SIGMA accurately detected 94.4% of scratching when the glove was donned. We believe that this simple device will empower dermatologists to more effectively measure and quantify itching and scratching in AD, and guide personalised treatment decisions.

Intranasal inorganic cerium oxide nanoparticles ameliorate oxidative stress induced motor manifestations in haloperidol-induced parkinsonism.

Cerium oxide nanoparticles (CONPs), owing to their radical scavenging property, have recently emerged as a therapeutic candidate for oxidative stress-mediated neurological diseases. However, oral and intravenous administration of CONPs is limited due to their poor physicochemical characteristics, low bioavailability, rapid systemic clearance, poor blood-brain penetration and dose-dependent toxicity. To overcome these challenges, we developed intranasal CONPs and evaluated their potential in the experimental PD model. CONPs were prepared by homogenous precipitation using tween 80 as a stabilizer and methanol/water as solvent. The optimization was done using Central Composite Design (CCD). The CONPs synthesis was confirmed by UV and FTIR. The optimized CONPs were small-sized (105.1 ± 5.78 nm), spherical (TEM), uniform (PDI, 0.119 ± 0.006) and stable (ZP, -22.7 ± 1.02 mV). Energy-dispersive X-ray analysis showed characteristic signals of Ce in developed CONPs. The X-ray diffraction pattern described the cubic fluorite structure and nano-crystalline nature of CONPs. The CONP anti-oxidant activity was found to be 93.60 ± 0.32% at 25 µg/mL concentration. Finally, motor manifestation studies like the forced swim test, locomotor test, akinesia, catalepsy, and muscle coordination test were conducted to assess the motor dysfunctions and behavioral activity in all four animal groups. Results of the in vivo motor manifestation studies in the haloperidol-induced PD rat model showed that co-administration of intranasal CONPs along with a half dose of levodopa resulted in significant protection, and results were significantly different from the untreated group but not significantly different from the healthy group. In conclusion, intranasal CONPs can be useful in ameliorating oxidative stress through their antioxidant effect and could be prospective therapeutics for the treatment of motor manifestations in Parkinson's disease.

Superparamagnetic Iron-Oxide Nanoparticles Synthesized via Green Chemistry for the Potential Treatment of Breast Cancer.

In the emerging field of nanomedicine, nanoparticles have been widely considered as drug carriers and are now used in various clinically approved products. Therefore, in this study, we synthesized superparamagnetic iron-oxide nanoparticles (SPIONs) via green chemistry, and the SPIONs were further coated with tamoxifen-conjugated bovine serum albumin (BSA-SPIONs-TMX). The BSA-SPIONs-TMX were within the nanometric hydrodynamic size (117 ± 4 nm), with a small poly dispersity index (0.28 ± 0.02) and zeta potential of -30.2 ± 0.09 mV. FTIR, DSC, X-RD, and elemental analysis confirmed that BSA-SPIONs-TMX were successfully prepared. The saturation magnetization (Ms) of BSA-SPIONs-TMX was found to be ~8.31 emu/g, indicating that BSA-SPIONs-TMX possess superparamagnetic properties for theragnostic applications. In addition, BSA-SPIONs-TMX were efficiently internalized into breast cancer cell lines (MCF-7 and T47D) and were effective in reducing cell proliferation of breast cancer cells, with IC50 values of 4.97 ± 0.42 µM and 6.29 ± 0.21 µM in MCF-7 and T47D cells, respectively. Furthermore, an acute toxicity study on rats confirmed that these BSA-SPIONs-TMX are safe for use in drug delivery systems. In conclusion, green synthesized superparamagnetic iron-oxide nanoparticles have the potential to be used as drug delivery carriers and may also have diagnostic applications.

Electronic nose as a tool for early detection of diseases and quality monitoring in fresh postharvest produce: A comprehensive review.

Postharvest diseases and quality degradation are the major factors causing food losses in the fresh produce supply chain. Hence, detecting diseases and quality deterioration at the asymptomatic stage of produce enables growers to treat the diseases earlier, maintain quality and reduce postharvest food losses. With the emergence of numerous technologies to detect diseases early and monitor the quality of fresh produce, such as polymerase chain reaction, gas chromatography-mass spectrophotometry, and near-infrared spectroscopy, electronic nose (EN) has also gained acknowledgement and popularity in the past decade as a robust and non-invasive analysis tool to detect odor profile and establish volatile biomarkers for metabolomics databases. However, literature reviewing the EN research on the early detection of diseases in produce after harvest is scarce. The fundamental concept of EN working principles (odor sampling, gas detection, and data acquisition method), as well as the application of EN as a whole, are covered in the first section of the review. An in-depth discussion of the application of EN analysis in the early identification of postharvest diseases and quality monitoring is provided in the subsequent sections, which is the key objective of this comprehensive review. The prospect, limitations, and likely future developments of EN in the postharvest sector are further highlighted in the last section.

Anatomy of the Sural Nerve in the Posterolateral Approach to the Ankle: A Cadaveric Study.

Sural nerve injury may occur during the posterolateral approach to the ankle during fracture fixation. We aimed to map its location in a posterolateral approach in cadaveric specimens. A posterolateral approach was used in 28 cadaver legs with the incision made halfway between the medial border of the fibula and the lateral border of Achilles tendon, extending proximally from the tip of the lateral malleolus. The sural nerve was identified and the distance from the distal tip of the incision to where it crossed the incision proximally was measured. The mean distance was 3.4 ± 1.2 (range 0.5-7.0) cm. In 22 cases (78.5%), the distance from the lowest part of the incision to the inferior part of the nerve was between 2.7 and 4.5 cm. The nerve did not cross the incision in 2 cases. We have demonstrated that the sural nerve crossed the posterolateral incision between 2.7 and 4.5 cm proximal to the tip of the fibula in the majority of cases. However, there remains individual anatomical variation, and we would recommend that care should be taken to look for the nerve closer to the Achilles tendon proximally and nearer the fibula distally. We hope that this information can help surgeons plan their approach and minimize iatrogenic injury to the sural nerve.

Evaluation of C4b as an adjunct marker in symptomatic RT-PCR negative Covid-19 cases.

Introduction: Detecting low viral load has been a challenge in this pandemic, which has led to its escalated transmission. Complement activation has been implicated in pathogenesis of Covid-19 infection. Thus, evaluation of complement activation in suspected Covid-19 infection may help to detect infection and limit false negative cases thus limiting transmission of infection. We speculate that measuring C4b, produced from an activated complement system due to the presence of Covid-19 may help in its detection, even when the viral titers are low. Methods: Plasma C4b levels of symptomatic RT-PCR positive patients (cases, n = 40); symptomatic RT-PCR negative patients (n = 35) and asymptomatic RT-PCR negative controls (n = 40) were evaluated. Plasma C5b-9, IL-6, D-dimer and C1-Inhibitor (C1-INH) were also measured in cases and controls. ELISA kits were used for all measurements. Statistical analyses were carried out using Stata, version 12 (Stata Corp., Texas, USA). Results: C4b levels were found to be significantly increased in RT-PCR positive patients as compared to asymptomatic RT-PCR negative controls. RT-PCR negative but symptomatic patients still showed increased C4b levels. The significantly higher levels of C4b in cases with a cut-off value of ≥ 116 ng/ml with optimum sensitivity and specificity of 80% and 52% respectively is indicative of its possible use as an adjunct marker. Increased levels of D-dimer, IL6, along with decreased levels of C1-INH were found in cases compared to controls. Whereas, C5b-9 levels were not significantly raised in cases. Conclusions: The results of our study suggests that plasma C4b may help to detect infection in false negative cases of RT-PCR that escape detection owing to low viral load. However, to confirm it a large-scale study is needed. Supplementary Information: The online version contains supplementary material available at 10.1007/s12291-022-01033-z.

Addressing cellular heterogeneity in tumor and circulation for refined prognostication.

Despite pronounced genomic and transcriptomic heterogeneity in non-small-cell lung cancer (NSCLC) not only between tumors, but also within a tumor, validation of clinically relevant gene signatures for prognostication has relied upon single-tissue samples, including 2 commercially available multigene tests (MGTs). Here we report an unanticipated impact of intratumor heterogeneity (ITH) on risk prediction of recurrence in NSCLC, underscoring the need for a better genomic strategy to refine prognostication. By leveraging label-free, inertial-focusing microfluidic approaches in retrieving circulating tumor cells (CTCs) at single-cell resolution, we further identified specific gene signatures with distinct expression profiles in CTCs from patients with differing metastatic potential. Notably, a refined prognostic risk model that reconciles the level of ITH and CTC-derived gene expression data outperformed the initial classifier in predicting recurrence-free survival (RFS). We propose tailored approaches to providing reliable risk estimates while accounting for ITH-driven variance in NSCLC.

Wearable Soft Microtube Sensors for Quantitative Home-Based Erectile Dysfunction Monitoring.

Quantifiable erectile dysfunction (ED) diagnosis involves the monitoring of rigidity and tumescence of the penile shaft during nocturnal penile tumescence (NPT). In this work, we introduce Erectile Dysfunction SENsor (EDSEN), a home-based wearable device for quantitative penile health monitoring based on stretchable microtubular sensing technology. Two types of sensors, the T- and R-sensors, are developed to effectively measure penile tumescence and rigidity, respectively. Conical models mimicking penile shaft were fabricated with polydimethylsiloxane (PDMS) material, using different base to curing agent ratios to replicate the different hardness properties of a penile shaft. A theoretical buckling force chart for the different penile models is generated to determine sufficiency criteria for sexual intercourse. An average erect penile length and circumference requires at least a Young's modulus of 179 kPa for optimal buckling force required for satisfactory sexual intercourse. The conical penile models were evaluated using EDSEN. Our results verified that the circumference of a penile shaft can be accurately measured by T-sensor and rigidity using the R-sensor. EDSEN provides a private and quantitative method to detect ED within the comfortable confines of the user's home.

Simultaneous estimation of paclitaxel and erlotinib in plasma by liquid chromatography/(+) electrospray tandem mass spectrometry: application in formulation development and pharmacokinetics.

The bio-analytical method was developed and validated for simultaneous detection and quantification of paclitaxel (PAC) and erlotinib (ERL) in plasma samples. The sample preparation process was accomplished by liquid-liquid extraction technique. The dried and reconstituted samples were subjected to chromatography on Discovery -C18 (50 × 4.6 × 5µm) column and a mobile phase, composed of a mixture of 0.1% formic acid in water: acetonitrile (70:30, v/v), in isocratic mode at a flow rate of 0.6 mL/min. Liquid chromatography coupled to tandem mass spectrometry detection in positive ion mode was selected to provide optimal selectivity and sensitivity. The mass transitions of erlotinib, erlotinib13C6, Paclitaxel and docetaxel were m/z 394.5→278.4, m/z 400.4→284.5, m/z 876.6→308.4 and m/z 830.0→304.0 respectively. The linearity in the calibration curves was obtained in the concentration range of 3.6 - 1006.7 ng/ml (r ≥ 0.99) for erlotinib and 5.3 - 1500.0 ng/mL for paclitaxel with an LLOQ (lower limit of quantification) of 3.6 and 5.3 ng/ml respectively. The run time was achieved in 2.5 min only, for all the analytes.

Ligand decorated biodegradable nanomedicine in the treatment of cancer.

Cancer is one of the major global health problems, responsible for the second-highest number of deaths. The genetic and epigenetic changes in the oncogenes or tumor suppressor genes alter the regulatory pathways leading to its onset and progression. Conventional methods are used in appropriate combinations for the treatment. Surgery effectively treats localized tumors; however, it fails to treat metastatic tumors, leading to a spread in other organs, causing a high recurrence rate and death. Among the different strategies, the nanocarriers-based approach is highly sought for, but its nonspecific delivery can cause a profound side effect on healthy cells. Targeted nanomedicine has the advantage of targeting cancer cells specifically by interacting with the receptors overexpressed on their surface, overcoming its non-specificity to target healthy cells. Nanocarriers prepared from biodegradable and biocompatible materials are decorated with different ligands by encapsulating therapeutic or diagnostic agents or both to target cancer cells overexpressing the receptors. Scientists are now utilizing a theranostic approach to simultaneously evaluate nanocarrier bio-distribution and its effect on the treatment regime. Herein, we have summarized the recent 5-year efforts in the development of the ligands decorated biodegradable nanocarriers, as a targeted nanomedicine approach, which has been highly promising in the treatment of cancer.

Biological control: a novel strategy for the control of the plant parasitic nematodes.

Plant parasitic nematodes (Root-knot nematodes, Meloidogyne spp.) are rounded worms, microscopic, and cause many agricultural economic losses. Their attacks have a direct impact on the productivity of cultivated crops by reducing their fruit quantity. Chemical control is widespread all over the world, but biological control is the most effective way to reduce the number of pests that infect crops, particularly by the use of microorganisms like fungi and bacteria. Biological control is rapidly evolving, and more products are being sold worldwide over time. They can be produced by fungi, bacteria, or actinomycetes that can destruct plant parasite nematodes and feed on them. Nematophagous microorganisms as the natural enemies of nematodes have a promising way of controlling nematodes. Some of them create net-like substances and traps to take the worms from outside and finally kill them. Other parasites serve as internal parasites in order to produce toxins and to produce virulence to kill nematodes. Comprehension of the molecular basis for microbial nematode interactions gives important insights into how successful biological nematode control agents can be created. We discuss recent advances in our understanding of nematodes and nematophagous microorganisms, with an emphasis on molecular mechanisms that infect nematodes with nematophagous microorganisms and on nematode safety from pathogenic attacks. Finally, we addressed numerous key areas for future research and development, including possible approaches to the application of our recent expertise in the development of successful biocontrol strategies.

Extraction of protein from food waste: An overview of current status and opportunities.

The chief intent of this review is to explain the different extraction techniques and efficiencies for the recovery of protein from food waste (FW) sources. Although FW is not a new concept, increasing concerns about chronic hunger, nutritional deficiency, food security, and sustainability have intensified attention on alternative and sustainable sources of protein for food and feed. Initiatives to extract and utilize protein from FW on a commercial scale have been undertaken, mainly in the developed countries, but they remain largely underutilized and generally suited for low-quality products. The current analysis reveals the extraction of protein from FW is a many-sided (complex) issue, and that identifies for a stronger and extensive integration of diverse extraction perspectives, focusing on nutritional quality, yield, and functionality of the isolated protein as a valued recycled ingredient.

Prospective Molecular Profiling of Circulating Tumor Cells from Patients with Melanoma Receiving Combinatorial Immunotherapy.

BACKGROUND: Blood molecular profiling of circulating tumor cells (CTCs) can enable monitoring of patients with metastatic melanoma during checkpoint inhibitor immunotherapy (CII) and in combination with targeted therapies. We developed a microfluidics-based CTC platform to explore CTC profiling utility in CII-treated patients with melanoma using a melanoma messenger RNA (mRNA)/DNA biomarker panel. METHODS: Blood samples (n = 213) were collected prospectively from 75 American Joint Committee on Cancer-staged III/IV melanoma patients during CII treatment and those enriched for CTCs. CTC profiling was performed using 5 known melanoma mRNA biomarkers and BRAF V600E DNA mutation. CTC biomarker status associations with clinical outcomes were assessed. RESULTS: CTCs were detected in 88% of blood samples from patients with melanoma. CTC-derived biomarkers and clinical variables analyzed using classification and regression tree analysis revealed that a combination of lactate dehydrogenase, CTC-mRNA biomarkers, and tumor BRAF-mutation status was indicative of clinical outcomes for patients with stage IV melanoma (n = 52). The panel stratified low-risk and high-risk patients, whereby the latter had poor disease-free (P = 0.03) and overall survival (P = 0.02). Incorporation of a DNA biomarker with mRNA profiling increased overall CTC-detection capability by 57% compared to mRNA profiling only. RNA sequencing of isolated CTCs identified significant catenin beta 1 (CTNNB1) overexpression (P <0.01) compared to nondisease donor blood. CTC-CTNNB1 was associated with progressive disease/stable disease compared to complete-responder patient status (P = 0.02). Serial CTC profiling identified subclinical disease in patients who developed progressive disease during treatment/follow-up. CONCLUSIONS: CTC-derived mRNA/DNA biomarkers have utility for monitoring CII, targeted, and combinatorial therapies in metastatic melanoma patients.

First report of postharvest stem end rot disease on MD2 pineapple fruits caused by Neoscytalidium dimidiatum in Malaysia.

Repeated sampling conducted from December 2019 to March 2020, and fruit of pineapple (Ananas comosus) var MD2 showing early stem end rot symptoms including brown and rotten fruit skin near the stem end region (Fig.1Aa) or darker skin with black discoloration (Fig.1Ab) indicated a consistent fungal infection. The samples (30 fruits from each location) were collected from store houses in three farmer fields with 60% disease incidence in Serdang, (3.0220oN,101.7055oE), Selangor, West Malaysia. The pulp of infected fruits appeared watery with characteristic spoilage odour. Symptomatic necrotic tissues from stem end region and skin were cut in to pieces (1x1cm), surface sterilized and plated onto potato dextrose agar amended aseptically with 0.5 g L-1 streptomycin sulphate. The plates were incubated at room temperature (28±2oC) in natural light conditions. Five days old cultures were light grey in colour and gradually turned dark brown to black with dense deeply tufted, mycelium as the culture aged (Fig.1B, C). Conidial morphology was observed using compound microscope (Olympus model BX-50F4, Tokyo, Japan) equipped with Dino-Eye. Branched mycelia with 0-1 septate arthospores were evident in 14 days old cultures (Fig.1D). Measured arthroconidia (5 to10x3 to 4.5µm) were ellipsoid to ovoid or round shaped, hyaline with an acutely rounded apex, truncate base, initially aseptate (Fig.1E) and arranged as chain at maturity (Fig.1F). The pathogen was identified through PCR amplification of the internal transcribed spacer (ITS) region using ITS1 and ITS4 primers (White et al., 1990) and BLASTn homology search as Neoscytalidium dimidiatum based on 100% similarity to a reference sequence (accession number KJ648577) that was previously deposited (Mohd et al.,2013). The sequence was deposited in Gen Bank ( accession number MW082810). Pathogenicity test was performed using the mycelial plug inoculation method and repeated twice with five replicates. Healthy MD2 pineapple fruits were surface sterilized with 1% NaOCl solution for15 min. followed by washing with sterilized distilled water. One centimeter diameter PDA plug at the margin of actively growing seven days old cultures were inserted in each of two inoculation wounds made on the skin and stem end of each fruit then the wounds were wrapped with moist cotton wool. Non-colonized PDA plugs were used to inoculate the control fruits. Fruits were incubated under 85% RH at room temperature. Five days after inoculation, the fruits showed similar dark necrotic discoloration and confirmed as N.dimidiatum by PCR (Fig.1G). The Koch postulates were fulfilled by inoculation and re-isolation of the fungal pathogen. This pathogen has also been reported previously to cause economic losses on a number of other hosts, such as pitayah fruits in Israel and Malaysia (Erza et al., 2013; Mohd et al., 2013)) and almond in California (Mohomed et al., 2018). To our knowledge this is the first report of N. dimidiatum causing postharvest stem end rot on MD2 pineapple in Malaysia. It may have the possibility to develop postharvest economic losses to pineapple industry, if severely affected fruits with high population of the pathogen left unattended in store houses.

Browning metabolism and quality of fresh-cut drumstick (Moringa oleifera) as influenced by acidulant treatments.

Though fresh-cut products save our time, but they are very much prone to enzymatic browning that drastically affects product's quality and marketability. Drumstick pods are considered as super food due to high nutritional contents. However, the fresh-cut pods are prone to brown discoloration. The enzyme activities promote the softening and cut-surface browning of pods, thus deteriorates their texture, decreases consumer appeal and shortens the shelf life. So, we aimed to assess the effect of citric (1%) and ascorbic (1%) acid treatments on quality attributes of fresh-cut drumsticks at 3-d interval during storage (5 ± 1 °C). In general there was an increase in lignin and quinone contents, while phenolic content was decreased during storage. However, samples subjected to ascorbic acid dip had higher phenolic content, lower rate of lignin formation, and reduced membrane permeability. Enzyme activities (polyphenol oxidase and peroxidase) were found to increase during storage, however, samples treated with ascorbic acid showed lower activities than that of the control and citric acid treated samples. The reduced enzyme activities resulted in the reduced browning incidence and maintained the quality. Therefore, postharvest dip of fresh-cut drumstick in to ascorbic acid (1%) could be suggested to increase the shelf life with reduced browning during low temperature storage.

Should the Tibiotalar Angle Be Measured Using an AP or Mortise Radiograph? Does It Matter?

Understanding the tibiotalar angle (TTA) is key to planning for deformity correction. The TTA is an important radiographic tool to determine alignment or malalignment of the ankle and hindfoot. Two methods of measuring the TTA have been described: the midline TTA (MTTA) and the lateral TTA (LTTA). The aim of this study was to compare the 2 angles as measured on mortise and anteroposterior (AP) radiographs in a series of normal and pathological cases. A radiographic review was performed of sequential ankle AP and mortise radiographs taken between January 2016 and September 2017 across 4 specialist orthopedic centers. Patients were categorized into a normal group, where patients had normal radiological appearances, and an arthritis group, where patients had radiographic arthritis. The MTTA and the LTTA were measured. The overall mean ± standard deviation MTTA was 88.7° ± 5.1°, and mean LTTA was 87.5° ± 5.2° (p < .01). There was no statistically significant difference between the MTTA and LTTA in the normal group or on AP radiographs alone (p = .09). There was a statistically significant difference between the MTTA and LTTA in the arthritis group (p < .01) and when measured on mortise radiographs (p = .02). The MTTA had no difference when measured on the AP and mortise radiographs. There was a statistically significant difference in the LTTA between AP and mortise radiographs (p = .04). We have shown the MTTA to be a reliable and reproducible tool in all patients, on AP and mortise radiographs. The type of radiograph does not alter the measurement of deformity. In contrast, we have shown the LTTA to be unreliable and statistically different when measured on AP and mortise radiographs.