Optimization, Purification, and Characterization of Haloalkaline Serine Protease from a Haloalkaliphilic Archaeon Natrialba hulunbeirensis Strain WNHS14

( Rania S Ahmed ),( Amira M Embaby ),( Mostafa Hassan ),( Nadia A Soliman ),( Yasser R Abdel-fattah ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 한국미생물·생명공학회지 Vol.49 No.2

The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The Box- Behnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

Expression of survivin in squamous cell carcinoma and transitional cell carcinoma of the urinary bladder: A comparative immunohistochemical study

Rania Makboul,Abeer EL-Refaiy M. Refaiy,Fatma Ahmed Mahmoud Badary,Islam F. Abdelkawi,Axel S. Merseburger,Rabab Ahmed Ahmed Mohammed 대한비뇨의학회 2015 Investigative and Clinical Urology Vol.56 No.1

Purpose: To compare the expression of survivin and its association with clinicopathological criteria in major types of urinary bladdercarcinoma, specifically, transitional cell carcinoma with and without squamous differentiation and squamous cell carcinoma. Materials and Methods: Immunohistochemical staining for survivin and Ki67 was performed on paraffin-embedded sections of104 carcinomas: 52 transitional cell carcinoma, 20 transitional cell carcinoma with squamous differentiation, and 32 squamous cellcarcinoma. Expression of survivin in >10% of tumor cells was described as altered survivin status. Ki67 staining in >20% of tumorcells was described as a high proliferation index. Results: Altered survivin expression was detected in 60/104 specimens (58%) and was significantly more frequent in transitionalcell carcinoma (78%) than in squamous cell carcinoma (38%) or transitional cell carcinoma with squamous differentiation (40%)(p<0.0001). In transitional cell carcinoma but not in squamous cell carcinoma, altered survivin status was associated with highertumor grade, higher proliferation index, and recurrence. In the whole specimens, altered survivin expression was significantly associatedwith advanced stage (p<0.001), recurrence (p=0.005), distant metastasis (p<0.001), and death (p=0.001). In the multivariateanalysis, altered survivin was an independent poor prognostic factor for recurrence. Conclusions: Unlike in transitional cell carcinoma, alteration of survivin expression in squamous cell carcinoma occurs less frequentlyand is not associated with features of tumor aggression or patient outcome. These findings raise a question: are urinarybladder carcinoma patients with squamous cell carcinoma type suitable candidates for survivin vaccine? This is an important questionto be answered before approving the vaccine in management.

Staphylococcus aureus derived hyaluronic acid and bacillus Calmette-Guérin purified proteins as immune enhancers to rabies vaccine and related immuno-histopathological alterations

Rania Ibrahim Shebl,Mohamed E. Amer,Tamer M. M. Abuamara,Emadeldin R. Matar,Hassan Fathy Ahmed,Tamer Albasyoni Gomah,Laila E. El Moselhy,Mohammed Abu-Elghait,Aly Fahmy Mohamed 대한백신학회 2021 Clinical and Experimental Vaccine Research Vol.10 No.3

Purpose: One of the essential goals regarding the successful control of rabies infection is the development of a safe, effective, and inexpensive vaccine. the current study aimed to evaluate the inactivation potential of β-propiolactone (βPL), binary ethyleneimine (BEI), and hydrogen peroxide (H2O2). Materials and Methods: Estimating the inactivation kinetics of βPL, BEI, and H2O2 revealed that the tested inactivants could completely and irreversibly inactivate rabies virus within 2, 12, and 4 hours, respectively while maintaining its viral immunogenicity. The potency of βPL, BEI, and H2O2 inactivated vaccines was higher than the World Health Organization acceptance limit and were in the order of 3.75, 4.21, and 3.64 IU/mL, respectively. Monitoring the humoral and cellular immunity elicited post-immunization using Staphylococcus aureus derived hyaluronic acid (HA) and bacillus Calmette-Guérin purified protein derivative (PPD) adjuvanted rabies vaccine candidates were carried out using enzyme-linked immunosorbent assay. Results: Results demonstrated that both adjuvants could progressively enhance the release of anti-rabies total immunoglobulin G as well as the pro-inflammatory mediators (interferon-gamma and interleukin-5) relative to time. However, a higher immune response was developed in the case of HA adjuvanted rabies vaccine compared to PPD adjuvanted one. The harmful consequences of the tested adjuvants were considered via investigating the histopathological changes in the tissues of the immunized rats using hematoxylin and eosin stain. Lower adverse effects were observed post-vaccination with HA and PPD adjuvanted vaccines compared to that detected following administration of the currently used alum as standard adjuvant. Conclusion: Our findings suggested that HA and PPD could serve as a promising platform for the development of newly adjuvanted rabies vaccines with elevated immune enhancing potentials and lower risk of health hazards.

Immunohistochemical Expression of Cytokeratins and Epithelial Membrane Protein 2 in Nasopharyngeal Carcinoma and its Potential Implications

Ahmed, Hussain Gadelkarim,Suliman, Rania Saad Abdul Gader,Abd El Aziz, Mohammed Siddig,Alshammari, Fawaz D. Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

Background: Nasopharyngeal carcinoma (NPC) is an aggressive disease and tends to involve surrounding tissues, and biomarkers for better management are yet to be identified. Materials and Methods: One hundred and fifty tissue samples with NPC diagnosis were were investigated using pan cytokeratin (CK) and epithelial membrane protein 2 (EMP2) antibodies. Results: Immunohistochemical expression of CK was identified in 144/150 (96%) and of EMP2 in 120/150 (80%). Conclusions: There is a high loss of EMP2 in NPC, especially high grade examples. Loss of CK expression is also linked to high grade NPC types.

Ascitic Fluid Calprotectin and Serum Procalcitonin as Accurate Diagnostic Markers for Spontaneous Bacterial Peritonitis

Ahmed Abdel-Razik,Nasser Mousa,Dina Elhammady,Rania Elhelaly,Rasha Elzehery,Sherif Elbaz,Mohamed Eissa,Niveen El-Wakeel,Waleed Eldars 거트앤리버 소화기연관학회협의회 2016 Gut and Liver Vol.10 No.4

Background/Aims: The diagnosis of spontaneous bacterial peritonitis (SBP) is based on a polymorphonuclear leukocytes (PMNs) exceeding 250/μL in ascitic fluid. The aim of the study was to evaluate serum procalcitonin and ascitic fluid calprotectin as accurate diagnostic markers for detecting SBP. Methods: Seventy-nine patients with cirrhotic ascites were included. They were divided into a SBP group, including 52 patients, and a non-SBP group of 27 patients. Serum procalcitonin, ascitic calprotectin, and serum and ascitic levels of tumor necrosis factor α (TNF-α) and interleukin 6 (IL6) were measured using an enzyme-linked immunosorbent assay. Results: Serum procalcitonin and ascitic calprotectin were significantly higher in SBP patients than in non-SBP patients. Significant increases in both serum and ascitic levels of TNF-α and IL-6 were observed in SBP patients versus nonSBP patients. At a cutoff value of 0.94 ng/mL, serum procalcitonin had 94.3% sensitivity and 91.8% specificity for detecting SBP. In addition, at a cutoff value of 445 ng/mL, ascitic calprotectin had 95.4% sensitivity and 85.2% specificity for detecting SBP. Both were positively correlated with ascitic fluid proteins, PMN count, TNF-α, and IL-6. Conclusions: According to our findings, determination of serum procalcitonin levels and ascitic calprotectin appears to provide satisfactory diagnostic markers for the diagnosis of SBP.

Effect of diets on the developmental rate of calliphorid fly of forensic importance Chrysomya megacephala (Fabricius, 1794)

El Hadi Mohamed Rania Ali,Galil Fahd Mohammed Abd Al,Al-Keridis Lamya Ahmed,Al-Shuraym Laila A.,Al-Mekhlafi Fahd A.,Alhag Sadeq K. 한국응용곤충학회 2021 Journal of Asia-Pacific Entomology Vol.24 No.3

The aim of this study was to compare the effect of diets on the morphological parameters of different devel opmental stages of blowflies Chrysomya megacephala (Fabricius, 1794). Adult C. megacephala flies were collected from a goat cadaver and reared in the laboratory when the max and min temperature were 32 and 24 ◦ C and the relative max and min humidity were ranged between 55 and 31% respectively. Blowflies at various feeding stages (first-, second-, and third-instar larvae) received different types of diets (meat, liver, fats, and mixed). The effects of the different diets on the life cycle duration and the morphological parameters of various stages were observed. The results revealed that the fat diet prolonged the post-mortem interval (PMI) up to 298.5 ± 1.15 hrs, whereas the mixed food diet shortened PMI to the 243.67 ± 0.12 hrs. Significant variation were observed at level (F = 740.71 df = 3P < 0.001). Furthermore, the mixed food diet significantly increased the morphological parameters (length, width, and weight) for almost all stages during the life cycle of blowflies. In contrast, the fat diet decreased these parameters to the lowest level. The present findings may provide a scientific basis for the determination of PMI.

Voluntary testing for COVID-19: perceptions and utilization among the inhabitants of Saudi Arabia

Ehab A. Abo-Ali,Ahmed Mousa,Rania Hussien,Shahad Mousa,Shayma Al-Rubaki,Mennatulla Omar,Badr Al-Haffashi,Abdullah Almilaibary 질병관리본부 2022 Osong Public Health and Research Persptectives Vol.13 No.3

Objectives: Voluntary testing (VT) plays a crucial role in the prevention and control of infectious diseases. The present study investigated the perceptions and utilization of VT services for coronavirus disease 2019 (COVID-19) among the inhabitants of Saudi Arabia.Methods: In total, 3,510 adult participants from all provinces of Saudi Arabia were recruited via a national online survey.Results: Of the 3,510 participants, 88.9% were aware of the testing services available to them and of those, more than half (59.5%) had used the VT services and 96.1% were satisfied with the services. Contact with a positive COVID-19 case was the top reason for accessing VT, while a lack of awareness about the availability of VT services was the top perceived limiting factor. A history of chronic health conditions, anxiety and/or depression, and previous symptoms suggestive of COVID-19 were found to be predictors of the utilization of VT services (odds ratio [OR] 1.55, 95% confidence interval [CI] 1.22−1.96; OR 1.48, 95% CI 1.16−1.88; and OR 3.31, 95% CI 2.77−3.95), respectively.Conclusion: The awareness of voluntary COVID-19 testing services was satisfactory among the Saudi Arabian population, but can be improved. Sociodemographic and health history predictors of the utilization of VT services were identified.

Intravenous versus perineural dexmedetomidine as adjuvant in adductor canal block for total knee arthroplasty

Abo-Zeid Salim Maha Ahmed,Bakrey Sherine,Elbadrawy Rania Elmohamady 대한마취통증의학회 2023 Korean Journal of Anesthesiology Vol.76 No.4

Background: The shivering effect after spinal anesthesia in total knee arthroplasty (TKA) is challenging for anesthesiologists. This study aimed to compare two administration routes of dexmedetomidine as a post-neuraxial shivering prevention measure and an adjunctive analgesic and sedative agent.Methods: Fifty-six patients were randomly allocated into two equal groups. The intravenous dexmedetomidine (IV dex) group received an IV infusion of 0.5 µg/kg dexmedetomidine diluted in 20 ml saline and an adductor canal block (ACB) consisting of 20 ml of 0.25% levobupivacaine and 1 ml saline. The adductor canal block dexmedetomidine (ACB dex) group received a 20 ml IV infusion of saline and an ACB consisting of 20 ml 0.25% levobupivacaine and 1 ml of 0.5 µg/kg dexmedetomidine.Results: The incidence of shivering 1 h post spinal anesthesia was equal in both groups (50%); however, the shivering grade was significantly lower in the IV dex group 1 h postoperatively. The onset of sensory block was significantly later in the IV dex group (22.14 ± 2.52 min) than in the ACB dex group (12 ± 3.31 min). Postoperative analgesic duration (h) was significantly longer in the ACB dex group (12.28 ± 4.47) compared to the IV dex group (9.28 ± 1.90). The sedation scores were also significantly higher in the IV dex group in the preoperative, intraoperative, and immediate postoperative periods.Conclusions: While perineural ACB dexmedetomidine had similar intraoperative anti-shivering with less sedative effects as IV dexmedetomidine, it was associated with both less shivering control and superior analgesia post-TKA under spinal anesthesia.

Synergistic impact of multiwalled carbon nanotubes on the properties of Ni-Mo thin-film via electrodeposition technique

Mona Hasan Gomaa,Zeinab Abdel Hamid,Magdy Ahmed Mahmoud Ibrahim,Rania Abd El Sttar,El-Said Helmy El- Mosallamy 한국화학공학회 2023 Korean Journal of Chemical Engineering Vol.40 No.5

Carbon nanotubes (CNTs) are the hardest and strongest materials due to their perfect mechanical properties and excellent chemical, electrical, and thermal characteristics. Therefore, CNTs are attractive candidates for the development of innovative multifunctional nanocomposites. The goal of the study was to synthesize and characterize NiMoCNT nanocomposite coatings onto steel substrates by electrodeposition technique to enhance the properties of the NiMo layer coating. The electrodeposition was carried out galvanostatically, and the percentage of MWCNT (wt%) in the composites was investigated under various working circumstances, including current density, pH, temperature, and CNTs concentration in the electroplating bath. Different techniques, such as X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and energy dispersive X-ray analysis (EDS), were used for the composite characterization. Moreover, the corrosion resistance and the nanocomposites’ mechanical characteristics were investigated. The results of the NiMo alloy show that the enhancement in current density decreases the Mo content from 41.8 wt% (at 3.5×10−2 Acm−2) to 31.06 wt% (at 6.5×10−2 Acm−2). On the other hand, the results proved that as the concentration of CNTs in the bath increases, the wt% of CNTs co-deposited in the NiMo matrix enhances, peaking at 22.36 wt% at 0.03 gL−1. Furthermore, the findings show that the Mo content of the coating is reduced when CNTs are present. In comparison to a NiMo coating without CNTs, the composite incorporating CNTs exhibits better corrosion resistance. In addition, the mechanical properties show that the microhardness of NiMoCNT composite-coated steel is better than that of NiMo, and the highest microhardness of NiMoCNT composite coated steel was 4.69 GPa, while pure NiMo coated steel had a microhardness of 2.37 GPa.

Growth and development of Chrysomya saffranea (Diptera: Calliphoridae) in response to different food substrates

Abd Al Galil Fahd Mohammed,Al‐Keridis Lamya Ahmed,AL‐Mekhlafi Fahd A.,Al‐Shuraym Laila A.,Alhag Sadeq K.,Ali El Hadi Mohamed Rania,Wadaan Muhammad A. 한국곤충학회 2021 Entomological Research Vol.51 No.10

Chrysomya saffranea is an insect that is important in criminal forensics as it plays a large role in decomposition on human cadavers. In this paper, the different developmental stages of C. saffranea flies were studied in the laboratory on different food, such as meat, liver, fat tissue and mixed food, to test whether these media affect the development of their larvae and have an impact on the calculation of the post-mortem interval. Our results indicated a significant growth enhancement among flies that fed on the mixed food (pure fat, meat and liver minced in equal proportions) compared with those that fed on minced (pure fat tissues, pure meat of and minced liver) of buffalo. The present results also confirmed the impact of food on the growth of C. saffranea, as well as the post-mortem interval PMI.