Prevalence of extensively drug-resistant gram negative bacilli in surgical intensive care in Egypt

Introduction: the prevalence of extensively drug resistant gram negative bacilli (XDR-GNB) is rapidly progressing; however in Egypt data are sparse. We conducted the present study to  quantify the incidence, risk factors and outcome of patients harboring XDR-GNB. Methods: a one year prospective study was done by collecting all the bacteriological reports  for cultures sent from the surgical intensive care unit, Cairo university teaching hospital.  XDR-GNB were defined as any gram negative bacilli resistant to three or more classes of  antimicrobial agents .Patients with XDR-GNB compared with those sustaining non extensively drug-resistant infection. A multivariate logistic regression model was created to identify independent predictors of multi-resistance. Results: during one-year study period, a total of 152 samples (65%) out of 234 gram negative  bacilli samples developed extensively drug resistant infection. XDR strains were significantly  higher in Acinetobacterspp (86%), followed by Pseudomonas (63%), then Proteus (61%),  Klebsiella (52%), and E coli (47%). Fourth generation cephalosporine (Cefipime) had the lowest susceptibility (10%) followed by third generation cephalosporines (11%), Quinolones (31%), Amikacin (42%), Tazobactam (52%), Carbapinems (52%), and colistin (90%).Relaparotomy was the only significant risk factor for acquisition of XDR infection. Conclusion: extensively drug-resistant gram negative infections are frequent in our ICU. This  is an alarming health care issue in Egypt which emphasizes the need to rigorously implement  infection control practices

Low cost detection of hepatitis C virus RNA in HCV infected patients by SYBR Green I real-time PCR

The prevalence of hepatitis C virus (HCV) is highest in Egypt compared to other countries. Nucleic acid amplification test (NAT) allows detection of HCV early during the course of infection. Unfortunately, NAT is more expensive than ELISA, thus its routine use as a screening tool for blood products or in clinical practice is quite limited. The aim of this study was to compare two common RT-PCR methods, TaqMan probe technique and SYBR Green method in quantitative detection of HCV RNA for diagnosis and follow up of HCV patients. Among the recruited 220 HCV patients, 154 (70%) were HCV-RNA positive by both the techniques, while 24 (10.9%) were negative by both techniques. On the other hand, 40 (18.2%) cases were HCV RNA positive only by SYBR Green technique, and 2 (0.9%) only by TaqMan probe technique. Forty (20.4%) of the 196 chronic HCV cases were HCV-RNA positive by SYBR Green but negative by TaqMan probe technique.Conclusion: This method is useful for rapid qualitative detection of HCV infection and particularly suitable for routine diagnostic applications.Keywords: HCV, PCR, SYBR Green 1, TaqMan probe, Viral loa

ENHANCED ENZYMATIC ACTIVITY OF STREPTOMYCES GRISEOPLANUS L-ASPARGINASE VIA ITS INCORPORATION IN AN OIL-BASED NANOCARRIER

Objective: L-asparaginase (L-asp) is a vital enzyme used as a therapeutic agent in combination with other drugs in the treatment of acute lymphoma, melanosarcoma and lymphocytic leukemia. Immobilization of enzymes through loading on nanoemulsion (NE) results in some advantages such as enhancing their stability and increasing their resistance to proteases. Aim of the present study is to formulate L-asp loaded nanoemulsion to enhance its efficiency and thermal stability. Methods: Nanoemulsion loaded with L-asp crude extract (specific activity 13.23U/mg protein) was prepared employing oleic acid as oil, tween 20/tween 80 as surfactants and propylene glycol (PG) as co-surfactant. L-asp loaded NE underwent several thermodynamic stability studies and the optimized formulae were further examined for their biochemical properties and thermal stability. Results The developed formulations were spherical in shape and their sizes were in the nanometric dimensions with negatively charged zeta potential values. Upon comparing the enzyme activity of L-asp loaded NE employing tween 20 (F1) or tween80 (F4) at different concentrations, the results revealed that F4 NE showed higher enzymatic activity [323 U/ml] compared to F1 NE [197 U/ml] at the same concentration. The nanosized immobilized L-asp was more stable in the pH range from 8 to 8.5 as compared to free L-asp. The immobilized enzyme preserved about 59.11% of its residual activity at 50 °C; while free L-asp preserved about 33.84%. Conclusion: In the view of these results, NE composed of oleic acid, tween 80 and PG represents a promising dosage form for enhancing the activity and stability of Streptomyces griseoplanus L-asp

HARNESSING THE ANTIOXIDANT PROPERTY OF CERIUM AND YTTRIUM OXIDE NANOPARTICLES TO ENHANCE MESENCHYMAL STEM CELL PROLIFERATION

Objective: This work was designed to explore if cerium oxide (CeO2) and yttrium oxide (Y2O3) nanoparticles as antioxidant agents could potentiate the proliferation of mesenchymal stem cells (MSCs) derived from human dental pulp (hDPSCs).Methods: Nanoparticles were characterized by transmission electron microscopy, particle size and zeta potential, X-ray diffraction, Fourier-transform infrared spectroscopy, and scanning electron microscope (SEM) along with energy-dispersive X-ray spectrometry. Furthermore, MSCs were isolated from human dental pulp, propagated and characterized by flow cytometry. Thereafter, the proliferative impact of the suggested nanoparticles on hDPSCs was investigated by 3-(4,5)-dimethylthiazol)-2,5-diphenyl tetrazolium bromide assay.Results: Different sizes (14.09–26.50 nm and 18.80–31.31 nm) for CeO2 and Y2O3 respectively, morphology, charges, and proliferative efficacy in hDPSCs were recorded for both nanoparticles.Conclusion: Generally speaking, the tested nanoparticles heightened the proliferative response of hDPSCs with the most prominent effect exerted by 15 μg/ml of CeO2 and 5 μg/ml of Y2O3. It is reasonable to assume that the antioxidant property of CeO2 and Y2O3 be involved in strengthening the proliferation process of hDPSCs

PHYSIOLOGICAL AND BIOCHEMICAL RESPONSES OF WHEAT (TRITICUM AESTIVUM L.) PLANTS TO POLYAMINES UNDER LEAD STRESS

Objective: The distribution, growth, development and productivity of wheat plants are greatly affected by various abiotic stresses such as lead (Pb) stress which become one of the most abundant toxic metal in the earth crust. Under the three applied polyamine (PAs) applications, the efficiency of wheat plants to tolerate Pb2+ stress in terms of growth and yield characteristics was noticed to varying degrees. Methods: The current study focused on the impact of 2.0 mM lead (Pb2+) on growth and performance of wheat plants before and after PAs applications. The sterilized seeds were soaked for 8 h at room temperature, either in distilled water (as a control), 0.25 mM spermine (Spm), 0.50 mM spermidine (Spd), or in 1.0 mM putrescine (Put). Results: Point out that, better growth and yield characteristics, chlorophyll “a” (Chl-a), chlorophyll “b” (Chl-b), soluble sugars, indoles, and enzymatic antioxidants (i.e., peroxidase (POX), catalase, ascorbate peroxidase, ascorbate oxidase, polyphenol oxidase, and glutathione reductase) and the enzyme α-amylase contents were obtained with seed soaking in 0.25 mM Spm, 0.50 mM Spd, or 1.0 mM Put than those generated with seed soaking in water under 2.0 mM Pb2+ stress. In contrast, the concentration of endogenous Pb2+ was significantly reduced. Conclusion: Among all tested PAs, 1.0 mM Put showed the best results and thus is recommended, as seed soaking, for wheat to grow well under Pb2+ stress

Sustainable release of propranolol hydrochloride laden with biconjugated-ufasomes chitosan hydrogel attenuates cisplatin-induced sciatic nerve damage in in vitro/in vivo evaluation

Peripheral nerve injuries significantly impact patients’ quality of life and poor functional recovery. Chitosan–ufasomes (CTS–UFAs) exhibit biomimetic features, making them a viable choice for developing novel transdermal delivery for neural repair. This study aimed to investigate the role of CTS–UFAs loaded with the propranolol HCl (PRO) as a model drug in enhancing sciatica in cisplatin-induced sciatic nerve damage in rats. Hence, PRO–UFAs were primed, embedding either span 20 or 60 together with oleic acid and cholesterol using a thin-film hydration process based on full factorial design (2(4)). The influence of formulation factors on UFAs’ physicochemical characteristics and the optimum formulation selection were investigated using Design-Expert(®) software. Based on the optimal UFA formulation, PRO–CTS–UFAs were constructed and characterized using transmission electron microscopy, stability studies, and ex vivo permeation. In vivo trials on rats with a sciatic nerve injury tested the efficacy of PRO–CTS–UFA and PRO–UFA transdermal hydrogels, PRO solution, compared to normal rats. Additionally, oxidative stress and specific apoptotic biomarkers were assessed, supported by a sciatic nerve histopathological study. PRO–UFAs and PRO–CTS–UFAs disclosed entrapment efficiency of 82.72 ± 2.33% and 85.32 ± 2.65%, a particle size of 317.22 ± 6.43 and 336.12 ± 4.9 nm, ζ potential of −62.06 ± 0.07 and 65.24 ± 0.10 mV, and accumulatively released 70.95 ± 8.14% and 64.03 ± 1.9% PRO within 6 h, respectively. Moreover, PRO–CTS–UFAs significantly restored sciatic nerve structure, inhibited the cisplatin-dependent increase in peripheral myelin 22 gene expression and MDA levels, and further re-established sciatic nerve GSH and CAT content. Furthermore, they elicited MBP re-expression, BCL-2 mild expression, and inhibited TNF-α expression. Briefly, our findings proposed that CTS–UFAs are promising to enhance PRO transdermal delivery to manage sciatic nerve damage

<i>Garcinia cambogia</i> phenolics as potent anti-COVID-19 agents:phytochemical profiling, biological activities, and molecular docking

COVID-19 is a disease caused by the coronavirus SARS-CoV-2 and became a pandemic in a critically short time. Phenolic secondary metabolites attracted much attention from the pharmaceutical industries for their easily accessible natural sources and proven antiviral activity. In our mission, a metabolomics study of the Garcinia cambogia Roxb. fruit rind was performed using LC-HRESIMS to investigate its chemical profile, especially the polar aspects, followed by a detailed phytochemical analysis, which led to the isolation of eight known compounds. Using spectrometric techniques, the isolated compounds were identified as quercetin, amentoflavone, vitexin, rutin, naringin, catechin, p-coumaric, and gallic acids. The antiviral activities of the isolated compounds were investigated using two assays; the 3CL-Mpro enzyme showed that naringin had a potent effect with IC50 16.62 &mu;g/mL, followed by catechin and gallic acid (IC50 26.2, 30.35 &mu;g/mL, respectively), while the direct antiviral inhibition effect of naringin confirmed the potency with an EC50 of 0.0169 &mu;M. To show the molecular interaction, in situ molecular docking was carried out using a COVID-19 protease enzyme. Both biological effects and docking studies showed the hydrophobic interactions with Gln 189 or Glu 166, per the predicated binding pose of the isolated naringin

Establishment of Gypsophila paniculata root culture for biomass, saponin, and flavonoid production

Baby’s breath (Gypsophila paniculata L.) roots are valuable as pharmaceuticals due to the content of triterpenoid saponins and other bioactive phytochemicals. However, the long root harvest period and fluctuation of these constituent’s content are among the constraints to traditional agricultural production. The present study shows an efficient and reliable liquid root culture of G. paniculata cv. ‘Perfecta’ established in a shake flask system using different auxins, media, and sugars. Hairy root (HR) induction in G. paniculata through Rhizobium rhizogenes-mediated transformation was also investigated. α-Naphthaleneacetic acid (NAA) was optimized for adventitious roots (AR) biomass, saponin, and flavonoid production at 1 or 2 mg/L compared to IBA. Full strength Gamborg’s medium (B5) recorded higher saponin content; however, the highest yield of total saponin and total flavonoids was achieved by full strength Murashige and Skoog's (MS) medium. Sucrose was more essential for root growth and accumulated total saponins and flavonoids rather than fructose and glucose. Polymerase chain reaction (PCR) analysis showed that G. paniculata HR carried rolC gene of R. rhizogenes A4 strain in its genome but not virD2 gene. Compared to non-transformed root, saponin content of leaf and stem-derived HR was 2.7 and 2.3-fold, while total flavonoid content was 2.1 and 2.0-fold, respectively. The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity was also higher in HR extracts than in non-transgenic roots. This study established an efficient protocol for G. paniculata root cultures for sustainable production of important natural saponins and flavonoids

Green synthesized extracts/Au complex of <i>Phyllospongia lamellosa</i>:unrevealing the anti-cancer and anti-bacterial potentialities, supported by metabolomics and molecular modeling

The anti-cancer and anti-bacterial potential of the Red Sea sponge Phyllospongia lamellosa in its bulk (crude extracts) and gold nanostructure (loaded on gold nanaoparticles) were investigated. Metabolomics analysis was conducted, and subsequently, molecular modeling studies were conducted to explore and anticipate the P. lamellosa secondary metabolites and their potential target for their various bioactivities. The chloroformic extract (CE) and ethyl acetate extract (EE) of the P. lamellosa predicted to include bioactive lipophilic and moderately polar metabolites, respectively, were used to synthesize gold nanoparticles (AuNPs). The prepared AuNPs were characterized through transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and UV–vis spectrophotometric analyses. The cytotoxic activities were tested against MCF-7, MDB-231, and MCF-10A. Moreover, the anti-bacterial, antifungal, and anti-biofilm activity were assessed. Definite classes of metabolites were identified in CE (terpenoids) and EE (brominated phenyl ethers and sulfated fatty amides). Molecular modeling involving docking and molecular dynamics identified Protein-tyrosine phosphatase 1B (PTP1B) as a potential target for the anti-cancer activities of terpenoids. Moreover, CE exhibited the most powerful activity against breast cancer cell lines, matching our molecular modeling study. On the other hand, only EE was demonstrated to possess powerful anti-bacterial and anti-biofilm activity against Escherichia coli. In conclusion, depending on their bioactive metabolites, P. lamellosa-derived extracts, after being loaded on AuNPs, could be considered anti-cancer, anti-bacterial, and anti-biofilm bioactive products. Future work should be completed to produce drug leads

Establishment of Gypsophila paniculata root culture for biomass, saponin, and flavonoid production

Baby’s breath (Gypsophila paniculata L.) roots are valuable as pharmaceuticals due to the content of triterpenoid saponins and other bioactive phytochemicals. However, the long root harvest period and fluctuation of these constituent’s content are among the constraints to traditional agricultural production. The present study shows an efficient and reliable liquid root culture of G. paniculata cv. ‘Perfecta’ established in a shake flask system using different auxins, media, and sugars. Hairy root (HR) induction in G. paniculata through Rhizobium rhizogenes-mediated transformation was also investigated. α-Naphthaleneacetic acid (NAA) was optimized for adventitious roots (AR) biomass, saponin, and flavonoid production at 1 or 2 mg/L compared to IBA. Full strength Gamborg’s medium (B5) recorded higher saponin content; however, the highest yield of total saponin and total flavonoids was achieved by full strength Murashige and Skoog's (MS) medium. Sucrose was more essential for root growth and accumulated total saponins and flavonoids rather than fructose and glucose. Polymerase chain reaction (PCR) analysis showed that G. paniculata HR carried rolC gene of R. rhizogenes A4 strain in its genome but not virD2 gene. Compared to non-transformed root, saponin content of leaf and stem-derived HR was 2.7 and 2.3-fold, while total flavonoid content was 2.1 and 2.0-fold, respectively. The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity was also higher in HR extracts than in non-transgenic roots. This study established an efficient protocol for G. paniculata root cultures for sustainable production of important natural saponins and flavonoids