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Fulltext Construction of an escherichia coli expression vector for the non-structural (ns)-1 protein of avian influenza virus H5N1

Agusta, Istiqomah, Jacinta Santhanam, Yap, Wei Boon

Jurnal Sains Kesihatan Malaysia, 2020;18(2):73-81.
MyJurnal

In the search for universal vaccine candidates for the prevention of avian influenza, the non-structural (NS)-1 protein of avian influenza virus (AIV) H5N1 has shown promising potential for its ability to effectively stimulate the host immunity. This study was aimed to produce a bacterial expression plasmid using pRSET B vector to harbour the NS1 gene of AIV H5N1 (A/Chicken/Malaysia/5858/2004 (H5N1)) for protein expression in Escherichia coli (E. coli). The NS1 gene (687 bp) was initially amplified by polymerase chain reaction (PCR) and then cloned into a pGEM-T Easy TA vector. The NS1 gene was released from pGEM-T-NS1 using EcoRI and XhoI restriction enzymes (RE). The pRSET B vector was also linearized using the same RE. The digested NS1 gene and linearized pRSET B were ligated using T4 DNA ligase to form the expression plasmid, pRSET B-NS1. The NS1 gene sequence in pRSET B-NS1 was confirmed by DNA sequencing. To prepare recombinant bacterial cells for protein expression in the future, pRSET B-NS1 was transformed into E. coli strain BL21 (DE3) by heat-shock. Colonies bearing the recombinant plasmid were screened using PCR. The DNA sequencing analysis revealed that the NS1 gene sequence was 97% homologous to that of AIV H5N1 A/Chicken/Malaysia/5858/2004 (H5N1). These results indicated that the NS1 gene of influenza A/Chicken/Malaysia/5858/2004 (H5N1) was successfully amplified and cloned into a pRSET B vector. Bacterial colonies carrying pRSET B-NS1 can be used for the synthesis of NS1-based influenza vaccine in the future and thereby aid in the prevention of avian influenza.

Matched MeSH terms: Heat-Shock Response
Fulltext A case report: Community-acquired Pseudomonas aeruginosa necrotizing fasciitis in a morbidly obese diabetic young man can be fatal

Mazlan MZ, Zainal Abidin H, Wan Hassan WMN, Nik Mohamad NA, Salmuna ZN, Ibrahim K, et al.

IDCases, 2020;22:e01001.
PMID: 33204633 DOI: 10.1016/j.idcr.2020.e01001

We present a case study of a 26-year-old morbidly obese man with a three-day history of right leg pain and swelling. The swelling was associated with low grade fever. He was alert and conscious upon presentation to the hospital. His physical examination showed gross swelling of the entire right lower limb with no systemic manifestations. There was no discharge and bullae from the swelling area of the leg. He had high blood sugar and was newly diagnosed with type 2 diabetes mellitus. He was diagnosed with necrotizing fasciitis. An intravenous imipenem-cilastatin 500 mg every 6 h together with clindamycin 900 mg every 8 h was started empirically. Extensive wound debridement was performed. The swab culture obtained intraoperatively grew Pseudomonas aeruginosa. He required an above knee amputation due to worsening infection despite wound debridement. Post-operatively, he developed acute kidney injury with severe metabolic acidosis, which required daily hemodialysis. However, the patient deteriorated due to septic shock with multi-organ failure, resulting in his death.

Matched MeSH terms: Shock, Septic
Fulltext Prenatal ultrasound heating impacts on fluctuations in haematological analysis of Oryctolagus cuniculus

Ahmad Zaiki FW, Md Dom S, Abdul Razak HR, Hassan HF

Quant Imaging Med Surg, 2013 Oct;3(5):262-8.
PMID: 24273744 DOI: 10.3978/j.issn.2223-4292.2013.10.05

Prenatal Ultrasound (US) is commonly used as a routine procedure on pregnant women. It is generally perceived as a safe procedure due to the use of non-ionizing radiation. However, the neurotoxicity of diagnostic prenatal US was detected to have a correlation with high susceptibility to early developing fetus. This research involved in vivo experimental model by using 3(rd) trimester pregnant Oryctolagus cuniculus and exposing them to US exposures for 30, 60, and 90 minutes at their gestational day (GD) 28-29. The output power and intensities, spatial peak temporal average intensity (ISPTA) of US were varied from 0.4 to 0.7 W and 0.13 to 0.19 W/cm(2) respectively were tested initially in free-field, water. Haematological analysis was carried out to detect any changes in blood constituents. Statistically significant differences were detected in red blood cell (RBC) count (P<0.001), haemoglobin (Hb) concentration (P<0.001) and also platelet (PLT) count (P<0.001) in newborn of Oryctolagus cuniculus. These findings indicate the possibility of US heating in causing defects on studied animal.

Matched MeSH terms: High-Energy Shock Waves
Fulltext Strongyloides Hyperinfection Syndrome in an Immunosuppressed Patient

Cheo, Seng Wee, Tan, Yee Ann, Low, Qin Jian

Borneo Journal of Medical Sciences, 2019;13(1):39-43.
MyJurnal

Strongyloides stercoralis is an intestinal nematode which is endemic in tropical and subtropical countries. The global prevalence of Strongyloides is unknown. Strongyloidiasis is found more frequently in the socioeconomically disadvantaged, in institutionalized populations, and in rural areas. The spectrum of disease varies. It may cause asymptomatic infection, mild eosinophilia or hyperinfection syndrome in the most severe form. Here we reported a case of Strongyloides hyperinfection syndrome in an immunosuppressed patient. This patient is a 54-year-old man with myasthenia gravis on long term azathioprine and prednisolone. He presented with fever associated with diarrhoea and was in septic shock. His blood culture was positive forKlebsiella pneumoniae.Strongyloides stercoralis larvae were detected in his sputum and stool sample. He was diagnosed to have Strongyloides hyperinfection and was treated with subcutaneous ivermectin. He recovered well. Our case demonstrated the association of Strongyloideshyperinfection with superimposed gram-negative sepsis as a consequence of prolonged immunosuppression. A high index of suspicion is needed in approaching patient with risk factors of hyperinfection syndrome.

Matched MeSH terms: Shock, Septic
A case of co-infection: First reported case of severe plasmodium knowlesi malaria and dengue co-infection in Sabah, Malaysia

Wong CK, Md Fuzi NH, Baherin MF, Lee HG

Med J Malaysia, 2020 03;75(2):171-172.
PMID: 32281602

We report a rare case of severe Plasmodium knowlesi malaria and dengue co-infection in a 36-year-old lady with hyperparasitaemia, metabolic acidosis, haemolysis and acute kidney injury. She was in shock requiring inotropic support and elective intubation. She had pericardial tamponade which necessitate pericardiocentesis to allow for haemodynamic stability during haemodialysis. She underwent haemodialysis, was ventilated for six days and stayed in hospital for 29 days. She was discharged home well with almost complete renal recovery. Physicians must have a high degree of suspicion for dengue co-infection in malaria patients with plasma leakage such as pericardial effusion to allow for prompt management.

Matched MeSH terms: Shock
Fulltext Antimicrobial activity and mode of action of terpene linalyl anthranilate against carbapenemase-producing Klebsiella pneumoniae

Yang SK, Yusoff K, Ajat M, Yap WS, Lim SE, Lai KS

J Pharm Anal, 2021 Apr;11(2):210-219.
PMID: 34012697 DOI: 10.1016/j.jpha.2020.05.014

Mining of plant-derived antimicrobials is the major focus at current to counter antibiotic resistance. This study was conducted to characterize the antimicrobial activity and mode of action of linalyl anthranilate (LNA) against carbapenemase-producing Klebsiella pneumoniae (KPC-KP). LNA alone exhibited bactericidal activity at 2.5% (V/V), and in combination with meropenem (MPM) at 1.25% (V/V). Comparative proteomic analysis showed a significant reduction in the number of cytoplasmic and membrane proteins, indicating membrane damage in LNA-treated KPC-KP cells. Up-regulation of oxidative stress regulator proteins and down-regulation of oxidative stress-sensitive proteins indicated oxidative stress. Zeta potential measurement and outer membrane permeability assay revealed that LNA increases both bacterial surface charge and membrane permeability. Ethidium bromide influx/efflux assay showed increased uptake of ethidium bromide in LNA-treated cells, inferring membrane damage. Furthermore, intracellular leakage of nucleic acid and proteins was detected upon LNA treatment. Scanning and transmission electron microscopies again revealed the breakage of bacterial membrane and loss of intracellular materials. LNA was found to induce oxidative stress by generating reactive oxygen species (ROS) that initiate lipid peroxidation and damage the bacterial membrane. In conclusion, LNA generates ROS, initiates lipid peroxidation, and damages the bacterial membrane, resulting in intracellular leakage and eventually killing the KPC-KP cells.

Matched MeSH terms: Heat-Shock Proteins
Fulltext Proteomic Analysis on Anti-Proliferative and Apoptosis Effects of Curcumin Analog, 1,5-bis(4-Hydroxy-3-Methyoxyphenyl)-1,4-Pentadiene-3-One-Treated Human Glioblastoma and Neuroblastoma Cells

Lee YQ, Rajadurai P, Abas F, Othman I, Naidu R

Front Mol Biosci, 2021;8:645856.
PMID: 33996900 DOI: 10.3389/fmolb.2021.645856

Curcumin analogs with excellent biological properties have been synthesized to address and overcome the poor pharmacokinetic profiles of curcumin. This study aims to investigate the cytotoxicity, anti-proliferative, and apoptosis-inducing ability of curcumin analog, MS13 on human glioblastoma U-87 MG, and neuroblastoma SH-SY5Y cells, and to examine the global proteome changes in these cells following treatment. Our current findings showed that MS13 induced potent cytotoxicity and anti-proliferative effects on both cells. Increased caspase-3 activity and decreased bcl-2 concentration upon treatment indicate that MS13 induces apoptosis in these cells in a dose- and time-dependent manner. The label-free shotgun proteomic analysis has defined the protein profiles in both glioblastoma and neuroblastoma cells, whereby a total of nine common DEPs, inclusive of glyceraldehyde 3-phosphate dehydrogenase (GAPDH), alpha-enolase (ENO1), heat shock protein HSP 90-alpha (HSP90AA1), Heat shock protein HSP 90-beta (HSP90AB1), Eukaryotic translation initiation factor 5A-1 (EFI5A), heterogenous nuclear ribonucleoprotein K (HNRNPK), tubulin beta chain (TUBB), histone H2AX (H2AFX), and Protein SET were identified. Pathway analysis further elucidated that MS13 may induce its anti-tumor effects in both cells via the common enriched pathways, "Glycolysis" and "Post-translational protein modification." Conclusively, MS13 demonstrates an anti-cancer effect that may indicate its potential use in the management of brain malignancies.

Matched MeSH terms: Heat-Shock Proteins
Montelukast Reduces the Risk of Dengue Shock Syndrome in Dengue Patients

Ahmad A, Waseem T, Butt NF, Randhawa FA, Malik U, Shakoori TA

Trop Biomed, 2018 Dec 01;35(4):1115-1122.
PMID: 33601858

A significant percentage of dengue patients develop Dengue Shock Syndrome (DSS) which is characterized by increased vascular permeability, circulatory failure and often death. Montelukast, a cysteinyl leukotriene receptor antagonist regulates vascular permeability and we hypothesized that it may be effective in protecting against DSS. An open label, parallel, randomized controlled trial (RCT) was thus carried out at Mayo Hospital, Department of Medicine, Lahore. A total of 200 patients of dengue fever were recruited and randomized into two groups. The group A was treated with Montelukast 10 mg once daily for 5 days along with general supportive treatment. Group B received the standard supportive treatment and served as the control group. The frequency of DSS was compared in the two groups by Chi square test. A binary logistic regression analysis was conducted to assess the effects of montelukast treatment on onset of DSS after adjusting for gender, age, white cell count, platelet count, haematocrit, serum alanine transaminase (ALT) and aspartate transaminase (AST). Relative risk (RR), absolute risk reduction (ARR), relative risk reduction (RRR) and numbers needed to treat (NNT) were calculated. Significance level was set at p0.001, Odds ratio=5.01, 95% CI=2.17-11.60) even after adjusting for confounders. Montelukast reduced the absolute risk (ARR=22%) and the relative risk (RRR=71%) of DSS in dengue fever. Numbers needed to treat were 4.55. We thus conclude that treatment with oral montelukast may protect patients of dengue fever from DSS and greatly reduce mortality.

Matched MeSH terms: Shock
Risks and burdens of third-generation cephalosporin resistant Enterobacteriaceae in neonatal sepsis

Thatrimontrichai A, Premprat N, Janjindamai W, Dissaneevate S, Maneenil G

Trop Biomed, 2020 Sep 01;37(3):663-670.
PMID: 33612780 DOI: 10.47665/tb.37.3.663

Third-generation cephalosporin resistant Enterobacteriaceae (TCRE) is a global concern especially in neonatal sepsis. We performed a secondary data analysis in a Thai neonatal intensive care unit to identify the risk factors for acquisition of TCRE sepsis and mortality of Enterobacteriaceae sepsis between 1991 and 2017. Multivariate logistic and Cox proportional regression were used for analysis. Numbers of neonates with TCRE and nonTCRE sepsis were 100 and 41 patients, respectively. Medians (interquartile ranges) of gestational age, birthweight, onset of sepsis and total hospital stay of neonates with Enterobacteriaceae sepsis were 32 (28, 38) weeks, 1670 (1025, 2750) grams, 11 (6, 25) days and 41 (22, 74) days, respectively. Univariate and multivariate analysis, neonates with TCRE sepsis were more likely to have birth asphyxia (adjusted odds ratio [aOR] = 2.6; 95% confidence interval [CI] 1.1-6.0; p = 0.02) and history of aminoglycoside exposure (aOR = 2.9; 95% CI 1.3- 6.7; p = 0.01). In-hospital case fatality rate from Enterobacteriaceae sepsis was 26% (36/141). In Cox regression, neonates with TCRE sepsis was not an independent risk of non-survivors, but septic shock (adjusted hazard ratio = 9.9; 95% CI 5.0-19.7, p < 0.001) increased 30-day mortality in the final model. Asphyxia and previous aminoglycoside consumption were risks of acquisition for neonatal TCRE sepsis while the burden was not a significant difference. Infection prevention and control must be strictly implemented in high multidrug-resistant area.

Matched MeSH terms: Shock, Septic
Fulltext Post-partum streptococcal toxic shock syndrome associated with necrotizing fasciitis

Chua WC, Mazlan MZ, Ali S, Che Omar S, Wan Hassan WMN, Seevaunnantum SP, et al.

IDCases, 2017;9:91-94.
PMID: 28725564 DOI: 10.1016/j.idcr.2017.05.002

We report a fatal case of post-partum streptococcal toxic shock syndrome in a patient who was previously healthy and had presented to the emergency department with an extensive blistering ecchymotic lesions over her right buttock and thigh associated with severe pain. The pregnancy had been uncomplicated, and the mode of delivery had been spontaneous vaginal delivery with an episiotomy. She was found to have septicemic shock requiring high inotropic support. Subsequently, she was treated for necrotizing fasciitis, complicated by septicemic shock and multiple organ failures. A consensus was reached for extensive wound debridement to remove the source of infection; however, this approach was abandoned due to the patient's hemodynamic instability and the extremely high risks of surgery. Both the high vaginal swab and blister fluid culture revealed Group A beta hemolytic streptococcus infection. Intravenous carbapenem in combination with clindamycin was given. Other strategies attempted for streptococcal toxic removal included continuous veno-venous hemofiltration and administration of intravenous immunoglobulin. Unfortunately, the patient's condition worsened, and she succumbed to death on day 7 of hospitalization.

Matched MeSH terms: Shock, Septic
Fulltext Enterobacter gergoviae peritonitis in a patient on chronic ambulatory peritoneal dialysis - first reported case

Anna Misya’il Abdul Rashid, Lim, Christopher Thiam Seong

Malaysian Journal of Medicine and Health Sciences, 2017;13(2):67-69.
MyJurnal

Enterobacter gergoviae is a gram negative rod-shaped opportunistic organism reported to cause urinary and respiratory tract infections, but peritonitis caused by this organism is unknown. We report a case of 50-year-old patient on peritoneal dialysis (PD) presented with Enterobacter gergoviae peritonitis with septic shock. Despite Intraperitoneal (IP) cloxacillin 250mg qid and IP ceftazidime 1gram q24h and subsequent escalation with IP amikacin 2mg/kg q24h and IP vancomycin 15mg/kg q24h within the next 48 hours, his peritonitis remained refractory and required catheter removal. Although Enterobacter gergoviae is naturally sensitive to aminoglycosides, carbapenems and quinolones, it reacts differently to the beta lactam antibiotics. Their resistance to third-generation cephalosporins is fast emerging and treatment with third-generation cephalosporins may cause AmpC-overproducing mutants. The majority of
Enterobacteriaceae, including Extended-spectrum beta-lactamases producers, remain susceptible to carbapenems. Our report provides an unfavourable course of E. gergoviae peritonitis likely due to acquired secondary drug resistance during the therapy period.

Matched MeSH terms: Shock, Septic
Genoproteomics-assisted improvement of Andrographis paniculata: toward a promising molecular and conventional breeding platform for autogamous plants affecting the pharmaceutical industry

Valdiani A, Talei D, Lattoo SK, Ortiz R, Rasmussen SK, Batley J, et al.

Crit Rev Biotechnol, 2017 Sep;37(6):803-816.
PMID: 28049346 DOI: 10.1080/07388551.2016.1260525

Andrographis paniculata (Burm. f.) Wall. ex Nees. (AP) is a hermaphroditic, self-compatible, and habitual inbreeding plant. Its main bioactive component is andrographolide, which is capable of inducing autophagic cell death in some human cancer cells and helps fight HIV/AIDS. Increasing the andrographolide content by investigating the genetic mechanisms controlling its biosynthesis in order to improve and develop high-yielding cultivars are the main breeding targets for AP. However, there might exist some limitations or barriers for crossability within AP accessions. Recently, this problem was addressed in AP by using a combination of crossbreeding and biotechnology-aided genetic methods. This review emphasizes that development of a breeding platform in a hard-to-breed plant, such as AP, requires the involvement of a broad range of methods from classical genetics to molecular breeding. To this end, a phenological stage (for example, flowering and stigma development) can be simplified to a quantitative morphological trait (for example, bud or stigma length) to be used as an index to express the highest level of receptivity in order to manage outcrossing. The outcomes of the basic crossability research can be then employed in diallel mating and crossbreeding. This review explains how genomic data could produce useful information regarding genetic distance and its influence on the crossability of AP accessions. Our review indicates that co-dominant DNA markers, such as microsatellites, are also capable of resolving the evolutionary pathway and cryptic features of plant populations and such information can be used to select the best breeding strategy. This review also highlights the importance of proteomic analysis as a breeding tool. In this regard, protein diversification, as well as the impact of normal and stress-responsive proteins on morphometric and physiological behaviors, could be used in breeding programs. These findings have immense potential for improving plant production and, therefore, can be regarded as prospective breeding platforms for medicinal plants that have an autogamous mode of reproduction. Finally, this review suggests that novel site-directed genome editing approaches such as TALENs (Transcription Activator-Like Effector Nucleases) and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein-9 nuclease) systems together with other new plant breeding technologies (NPBT) should simultaneously be taken into consideration for improvement of pharmaceutical plants.

Matched MeSH terms: Heat-Shock Proteins
Fulltext Clinical Manifestations and Risk Factors of Streptococcus suis Mortality Among Northern Thai Population: Retrospective 13-Year Cohort Study

Rayanakorn A, Katip W, Goh BH, Oberdorfer P, Lee LH

Infect Drug Resist, 2019;12:3955-3965.
PMID: 32021313 DOI: 10.2147/IDR.S233326

Purpose: Streptococcus suis (S. suis) is an emerging zoonotic disease mainly in pigs, causing serious infections in humans with high prevalence in Southeast Asia. Despite a relatively high mortality rate, there are limited data regarding the risk factors of this life-threatening infection. Therefore, a 13-year retrospective cohort study in Chiang Mai, Thailand during 2005-2018 was conducted to explore risk factors associated with S. suis mortality and to update the outcomes of the disease.
Patients and methods: S. suis positive cases were derived from those with positive S. suis isolates from microbiological culture results and Matrix-Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF). Potential risk factors of mortality were identified using univariate and multivariate logistic regression.
Results: Of 133 patients with culture-proven S. suis infection identified, there were 92 males and 41 females. The mean age was 56.47 years. Septicemia (55.64%) was the most common clinical manifestation followed by meningitis (37.59%) and infective endocarditis (25.56%). Alcohol drinking and raw pork consumption were documented in 66 (49.62%) and 49 (36.84%) cases respectively. The overall mortality rate was 12.03% (n=16). According to the multivariate analysis, the independent risk factors for mortality were prolonged bacteremia ≥ 6 days (OR = 43.57, 95% CI = 2.46-772.80, P =0.010), septic shock (OR = 13.34, 95% CI = 1.63-109.03, P =0.016), and direct bilirubin > 1.5 mg/dL (OR = 12.86, 95% CI = 1.91-86.59, P =0.009).
Conclusion: S. suis is not infrequent in Northern Thailand, where the cultural food habit of raw pork eating is still practiced. To the best of our knowledge, this is the largest series focusing on risk factors of S. suis mortality which has been conducted in Thailand. Prolonged bacteremia ≥ 6 days, septic shock, and direct bilirubin > 1.5 mg/dL were strong predictors associated with S. suis mortality. The mortality risk factors identified may be further utilized in clinical practice and future research to improve patient outcomes.

Matched MeSH terms: Shock, Septic
Fulltext Management of Takotsubo Syndrome: A Comprehensive Review

Sattar Y, Siew KSW, Connerney M, Ullah W, Alraies MC

Cureus, 2020 Jan 03;12(1):e6556.
PMID: 32042529 DOI: 10.7759/cureus.6556

Takotsubo syndrome (TTS), also known as Takotsubo cardiomyopathy, is a transient left ventricular wall dysfunction that is often triggered by physical or emotional stressors. Although TTS is a rare disease with a prevalence of only 0.5% to 0.9% in the general population, it is often misdiagnosed as acute coronary syndrome. A diagnosis of TTS can be made using Mayo diagnostic criteria. The initial management of TTS includes dual antiplatelet therapy, anticoagulants, beta-blockers, angiotensin-converting enzyme inhibitors or aldosterone receptor blockers, and statins. Treatment is usually provided for up to three months and has a good safety profile. For TTS with complications such as cardiogenic shock, management depends on left ventricular outflow tract obstruction (LVOTO). In patients without LVOTO, inotropic agents can be used to maintain pressure, while inotropic agents are contraindicated in patients with LVOTO. In TTS with thromboembolism, heparin should be started, and patients should be bridged to warfarin for up to three months to prevent systemic emboli. Our comprehensive review discussed the management in detail, derived from the most recent literature from observational studies, systematic review, and meta-analyses.

Matched MeSH terms: Shock, Cardiogenic
Septic miscarriage with toxic shock syndrome and disseminated intravascular coagulation (DIC): The role of surgery, recombinant activated factor VII and intravenous immunoglobulin (IVIG)

Tan LN, Mariappa G, Voon HY, Suharjono H

Med J Malaysia, 2017 12;72(6):380-381.
PMID: 29308782 MyJurnal

Severe sepsis with multi-organ failure is associated with a high mortality rate. This case report highlights the challenges and modalities available in the management of a lady with refractory shock and disseminated intravascular coagulation (DIC) due to toxic shock syndrome (TSS) from genital tract sepsis. Early surgical intervention to remove the source of infection, the use of recombinant activated factor VII to treat intractable disseminated intravascular coagulation and intravenous immunoglobulin to neutralise the circulating exotoxins, have been employed and shown to drastically improve outcomes.

Matched MeSH terms: Shock, Septic
Effect of Gibberellin and Heat Shock on the Lipid Composition of Endoplasmic Reticulum in Barley Aleurone Layers

Grindstaff KK, Fielding LA, Brodl MR

Plant Physiol, 1996 Feb;110(2):571-581.
PMID: 12226205

The heat-shock responses of barley (Hordeum vulgare L. cv Hi- malaya) aleurone layers incubated with or without gibberellic acid (GA3) were compared. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that heat shock blocked the synthesis and secretion of secretory proteins from GA3-treated layers but not untreated layers. This suppression of secretory protein synthesis has been correlated with changes in endoplasmic reticulum (ER) membranes (F.C. Belanger, M. R. Brodl, T.-h.D. Ho [1986] Proc Natl Acad Sci USA 83: 1354-1358; L. Sticher, A.K. Biswas, D.S. Bush, R.L. Jones [1990] Plant Physiol 92: 506-513). Our secretion data suggested that the ER membranes of aleurone layers incubated without GA3 may be more heat shock tolerant. To investigate this, the lipid profiles of membrane extracts in aleurone layers labeled with [14C]glycerol were examined. Heat shock markedly increased [14C]glycerol incorporation into phosphatidylcholine (PC), and gas chromatography revealed an increase in the amount of saturated fatty acids associated with thin layer chromatography-purified PC in GA3-treated layers. In contrast, aleurone layers incubated without GA3 at normal temperature contained PC-associated fatty acids with a greater degree of saturation than GA3-treated layers. Heat shock modestly increased the degree of fatty acid saturation in untreated aleurone layers. This same trend was noted in fatty acids isolated from ER membranes purified by continuous sucrose density centrifugation. We propose that increased fatty acid saturation may help sustain ER membrane function in heat-shocked aleurone layers incubated in the absence of GA3.

Matched MeSH terms: Heat-Shock Response
Fulltext Gastroprotective activity of Annona muricata leaves against ethanol-induced gastric injury in rats via Hsp70/Bax involvement

Moghadamtousi SZ, Rouhollahi E, Karimian H, Fadaeinasab M, Abdulla MA, Kadir HA

Drug Des Devel Ther, 2014;8:2099-110.
PMID: 25378912 DOI: 10.2147/DDDT.S70096

The popular fruit tree of Annona muricata L. (Annonaceae), known as soursop and graviola, is a widely distributed plant in Central and South America and tropical countries. Leaves of A. muricata have been reported to possess antioxidant and anti-inflammatory activities. In this study, the gastroprotective effects of ethyl acetate extract of A. muricata leaves (EEAM) were investigated against ethanol-induced gastric injury models in rats. The acute toxicity test of EEAM in rats, carried out in two doses of 1 g/kg and 2 g/kg, showed the safety of this plant, even at the highest dose of 2 g/kg. The antiulcer study in rats (five groups, n=6) was performed with two doses of EEAM (200 mg/kg and 400 mg/kg) and with omeprazole (20 mg/kg), as a standard antiulcer drug. Gross and histological features showed the antiulcerogenic characterizations of EEAM. There was significant suppression on the ulcer lesion index of rats pretreated with EEAM, which was comparable to the omeprazole effect in the omeprazole control group. Oral administration of EEAM to rats caused a significant increase in the level of nitric oxide and antioxidant activities, including catalase, glutathione, and superoxide dismutase associated with attenuation in gastric acidity, and compensatory effect on the loss of gastric wall mucus. In addition, pretreatment of rats with EEAM caused significant reduction in the level of malondialdehyde, as a marker for oxidative stress, associated with an increase in prostaglandin E2 activity. Immunohistochemical staining also demonstrated that EEAM induced the downregulation of Bax and upregulation of Hsp70 proteins after pretreatment. Collectively, the present results suggest that EEAM has a promising antiulcer potential, which could be attributed to its suppressive effect against oxidative damage and preservative effect toward gastric wall mucus.

Matched MeSH terms: HSP70 Heat-Shock Proteins/metabolism*
Fulltext α-Mangostin from Cratoxylum arborescens demonstrates apoptogenesis in MCF-7 with regulation of NF-κB and Hsp70 protein modulation in vitro, and tumor reduction in vivo

Ibrahim MY, Hashim NM, Mohan S, Abdulla MA, Kamalidehghan B, Ghaderian M, et al.

Drug Des Devel Ther, 2014;8:1629-47.
PMID: 25302018 DOI: 10.2147/DDDT.S66105

Cratoxylum arborescens is an equatorial plant belonging to the family Guttiferae. In the current study, α-Mangostin (AM) was isolated and its cell death mechanism was studied. HCS was undertaken to detect the nuclear condensation, mitochondrial membrane potential, cell permeability, and the release of cytochrome c. An investigation for reactive oxygen species formation was conducted using fluorescent analysis. To determine the mechanism of cell death, human apoptosis proteome profiler assay was conducted. In addition, using immunofluorescence and immunoblotting, the levels of Bcl-2-associated X protein (Bax) and B-cell lymphoma (Bcl)-2 proteins were also tested. Caspaces such as 3/7, 8, and 9 were assessed during treatment. Using HCS and Western blot, the contribution of nuclear factor kappa-B (NF-κB) was investigated. AM had showed a selective cytotoxicity toward the cancer cells with no toxicity toward the normal cells even at 30 μg/mL, thereby indicating that AM has the attributes to induce cell death in tumor cells. The treatment of MCF-7 cells with AM prompted apoptosis with cell death-transducing signals. This regulated the mitochondrial membrane potential by down-regulation of Bcl-2 and up-regulation of Bax, thereby causing the release of cytochrome c from the mitochondria into the cytosol. The liberation of cytochrome c activated caspace-9, which, in turn, activated the downstream executioner caspace-3/7 with the cleaved poly (ADP-ribose) polymerase protein, thereby leading to apoptotic alterations. Increase of caspace 8 had showed the involvement of an extrinsic pathway. This type of apoptosis was suggested to occur through both extrinsic and intrinsic pathways and prevention of translocation of NF-κB from the cytoplasm to the nucleus. Our results revealed AM prompt apoptosis of MCF-7 cells through NF-κB, Bax/Bcl-2 and heat shock protein 70 modulation with the contribution of caspaces. Moreover, ingestion of AM at (30 and 60 mg/kg) significantly reduced tumor size in an animal model of breast cancer. Our results suggest that AM is a potentially useful agent for the treatment of breast cancer.

Matched MeSH terms: HSP70 Heat-Shock Proteins/metabolism*
Fulltext Gastroprotection studies of Schiff base zinc (II) derivative complex against acute superficial hemorrhagic mucosal lesions in rats

Golbabapour S, Gwaram NS, Hassandarvish P, Hajrezaie M, Kamalidehghan B, Abdulla MA, et al.

PLoS One, 2013;8(9):e75036.
PMID: 24058648 DOI: 10.1371/journal.pone.0075036

The study was carried out to assess the gastroprotective effect of the zinc (II) complex against ethanol-induced acute hemorrhagic lesions in rats.

Matched MeSH terms: HSP70 Heat-Shock Proteins/biosynthesis
Eurycomanone suppresses expression of lung cancer cell tumor markers, prohibitin, annexin 1 and endoplasmic reticulum protein 28

Wong PF, Cheong WF, Shu MH, Teh CH, Chan KL, AbuBakar S

Phytomedicine, 2012 Jan 15;19(2):138-44.
PMID: 21903368 DOI: 10.1016/j.phymed.2011.07.001

Bioactive compounds from the medicinal plant, Eurycoma longifolia Jack have been shown to promote anti-proliferative effects on various cancer cell lines. Here we examined the effects of purified eurycomanone, a quassinoid found in Eurycoma longifolia Jack extract, on the expression of selected genes of the A549 lung cancer cells. Eurycomanone inhibited A549 lung cancer cell proliferation in a dose-dependent manner at concentrations ranging from 5 to 20 μg/ml. The concentration that inhibited 50% of cell growth (GI(50)) was 5.1 μg/ml. The anti-proliferative effects were not fully reversible following the removal of eurycomanone, in which 30% of cell inhibition still remained (p<0.0001, T-test). At 8 μg/ml (GI(70)), eurycomanone suppressed anchorage-independent growth of A549 cells by >25% (p<0.05, T-test, n=8) as determined using soft agar colony formation assay. Cisplatin, a chemotherapy drug used for the treatment of non small cell lung cancer on the other hand, inhibited A549 cells proliferation at concentrations ranging from 0.2 μg/ml to 15 μg/ml with a GI(50) of 0.58 μg/ml. The treatment with eurycomanone reduced the abundance expression of the lung cancer markers, heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1, p53 tumor suppressor protein and other cancer-associated genes including prohibitin (PHB), annexin 1 (ANX1) and endoplasmic reticulum protein 28 (ERp28) but not the house keeping genes. The mRNA expressions of all genes with the exception of PHB were significantly downregulated, 72 h after treatment (p<0.05, T-test, n=9). These findings suggest that eurycomanone at viable therapeutic concentrations of 5-20 μg/ml exhibited significant anti-proliferative and anti-clonogenic cell growth effects on A549 lung cancer cells. The treatment also resulted in suppression of the lung cancer cell tumor markers and several known cancer cell growth-associated genes.

Matched MeSH terms: Heat-Shock Proteins/chemistry*

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