Home > Molecular Characterization of New Delhi Metallo-Β-Lactamase Ndm Producing Gram Negative Rods and the Efficacy of Manuka Honey Against Ndm Strain in Mouse Model
Molecular Characterization of New Delhi Metallo-Β-Lactamase Ndm Producing Gram Negative Rods and the Efficacy of Manuka Honey Against Ndm Strain in Mouse Model
New Delhi Metallo-β-lactamase (NDM) belongs to the B1 super family of metallo-β-lactamase and is rapidly disseminated globally. NDM producing pathogens confer resistance against a wide range of antibiotics including carbapenems; considered to be the last resort to treat such pathogens. Currently, therapeutic treatment of infections by these bacteria is becoming a big challenge. Manuka honey is the one that is widely used medicinal honey for wound dressing worldwide. Several factors are responsible for their antibacterial activity including high osmolality, pH, hydrogen peroxide concentration, phytochemical nature and methylglyoxal. Therefore, we planned to determine the molecular characterization of NDM producing bacteria and in vitro and in vivo efficacy of Manuka honey against NDM positive strain. A total of 302 carbapenem resistant Gram negative bacteria were collected from four different tertiary care hospitals Lahore. Isolates were reconfirmed by Vitek 2 system and MALDI-TOF. Carbapenamases and MBLs detection was done as per CLSI guidelines 2015. Bacteria were further analyzed for the presence of blaNDM by PCR and NDM variants were detected after Sanger sequencing. MIC (μg/mL) of the NDM positive bacteria were done by Vitek 2 system. MLST of NDM positive K. pneumoniae and E. coli was done by amplifying and sequence their house keeping genes. Plasmids characterization of NDM producing bacteria was done by S1 nuclease PFGE and In Gel DNA-DNA hybridization. In vitro activity of Manuka honey was done by screening the NDM positive bacteria by agar well diffusion method. MIC and MBC (%v/v) of Manuka honey against NDM positive pathogens was done by microbroth dilution method. In vivo activity of Manuka honey was done using BALB/c mouse model by intravenous injection of 20% and 30% of Manuka honey against NDM-1 positive K. pneumoniae strain. Bacterial load (CFU) was counted using Miles and Misra method after seven and fifteen days of clinical trial. Out of 302 clinical isolates, K. pneumoniae (29.8%) were identified followed by A. baumannii (28%) and E. coli (11%) in four tertiary care hospitals. Overall, highest prevalence (38.7%) of carbapenem resistant bacteria was observed in the Children hospital and culprit pathogen were K. pneumoniae (31.6%). Generally, 26.5% of the carbapenem resistant bacteria were recovered from blood culture, 25.4% from urine culture and 22.8% from pus swabs. Among these pathogens, 91% were carbapenamase and 89% were MBLs producers. After molecular identification, 144 (47.6%) bacteria were positive for NDM with majority of K. pneumoniae (52%) and E. coli (15.2%). Multi-sequence alignment revealed four different variants of NDM (NDM-1, 4, 5, and 7) and mainly these variants were found in K. pneumoniae and E. coli with the majority of NDM-1 (82.6%). Overall 100% of the NDM positive bacteria showed highest MIC (μg/mL) against commonly used antibiotics including meropenem, 70%-80% to moxifloxacin, aztreonam, cefepime and levofloxacin while most effective drugs were colistin and tigecycline. Most of the K. pneumoniae belonged to the sequence type (ST) of ST11, ST147 and ST29 however majority of the E. coli belonged to ST405 and ST101. Plasmids characterization displayed most of the isolates (n=130; 90%) carried blaNDM on their plasmids and 6 (4%) carried on chromosome however about 17% of the blaNDM carried on ~150kb of plasmids. Overall, in vitro activity of Manuka honey against 144 isolates exhibited that 55 (38%) NDM positive bacteria were inhibited at the concentration of 30% (v/v), 52 (36%) at 25% (v/v) and 30 (20.8%) at 20% (v/v). Highest MIC was observed against K. pneumoniae (35% v/v) and least observed to C. freundii and M. morganii (15% v/v). In vivo activity of Manuka honey against K. pneumoniae ST11 showed that there was a substantial reduction in the mean log value of each honey treated group as compared to the positive control group (Group B). The mean log value for Group C and D significantly and gradually reduced from 1.829 to 1.035 and 1.624 to 0.610 in first and second week respectively. Statically no significant difference was observed between the treatment Group C and D. This study concluded that there is an alarming spread of blaNDM and its variants in the Gram-negative bacteria particularly among children that displayed resistance to most of the drugs including carbapenems which can lead to treatment failure and mortality. In vitro Manuka honey has the potential effect against the NDM producing pathogens however in vivo the therapeutic effect of Manuka honey gradually and effectively reduces the bacterial load of K. pneumoniae ST11. Therefore, Manuka honey could be used as one of the remedy to treat such pathogens following several in vitro and in vivo clinical studies. Hence, it is need of the hour to conduct a national multicenter survey to combat this serious threat in our hospitals and also extensively look to treat these pathogens with Manuka honey.
[اہلیہ،مولانا سعیداحمد اکبرآبادی] میری اہلیہ!اختری بیگم کے انتقال پرملال پرجو دو ماہ کی شدید علالت کے بعد ۳۰ / شعبان المعظم مطابق ۱۴/جولائی کوعصر اورمغرب کے درمیان ہوا۔جن دوستوں، عزیزوں نے برصغیر ہندوپاک اوربیرونی ممالک سے تعزیت کے خطوط و ٹیلگرام بھیجے ہیں اورہندوستان وپاکستان کے جن اخبارات نے تعزیتی نوٹ کے ساتھ اس خبر کوشائع کیاہے اورجن اداروں نے اپنے یہاں اجتماعی ایصال ثواب کااہتمام کیا ہے ان سب کی محبت وہمدردی کاتہہ دل سے نہایت شکرگذار ہوں اوردعاکرتا ہوں جزاھم اﷲ عنی جزاء خیرا۔ ساتھ ہی معذرت اس کی کرنی ہے کہ اس حادثہ نے ذہن وقلم کوجیسے مفلوج کردیاہے، چنانچہ برہان کے زیرنظر شمارہ کے لیے جب نظرات بھی نہیں جاسکے تواپنے ناتمام مضامین کے فائل سے ایک مضمون نکال کر بھرت پورا کر دیا ہے ، اس لیے قارئین کلکتہ کے سفرنامہ اورتبصروں کے لیے معذور تصور فرمائیں۔ [سعید اکبر آبادی، ۷/اگست۱۹۸۰ء]
Organ transplantation and organ donation are an urgent problem of our time. This was done in different ways. The medical field introduces new treatments, including organ transplantation and service to humanity. It is allowed to donate organs both in Judaism and Christianity. What is allowed in the Islamic perspective, transplantation and organ donation only if they follow the goals of Islamic sharia? The human body has the honor and dignity that must be preserved in every movement of life. In the case of organ transplants, if it leads to the destruct-tion of a human being and the violation of his honor or paves the way for the abduction, which offers immorality, and in these circumstances, it should not be done. Organ donation and transplantation will be beneficial, under the supervision of a qualified transplant team. In this article, the permissibility of organ transpla-ntation in the light of Islam has been discussed.
This thesis is dedicated to theoretical characterizing of two component metamaterials as arrays of metal rods/wires periodically immersed in a dielectric isotropic matrix. The rods material is supposed to be non-magnetic. Only a circular cross section case of the rods is considered in the thesis while the rods do not touch each other. At the same time, there is no restriction to the radius of cross section of the rods. The microwave frequency range (from 0 to 5 GHz) has only been considered in this study. The metamaterial media/structures are being considered in the thesis as artificial semiconductors with their own effective complex dielectric and magnetic constants. So, the presented metamaterial media/structures in the thesis are considered as perfect crystals with their own dispersive properties studied in the microwave frequency range. The lattice constant of the crystal is equal to the constant of unit cell of metamaterial under consideration. The characterizations of considered metamaterial structures in the thesis are identified with the study of properties of the effective dielectric and magnetic constants as functions of the frequency (in the GHz frequencies) of incident electromagnetic wave and the volume fraction of metal rods in the unit cell. The above characterizing is the key for defining unusual material properties unavailable in real nature: enhancement of the effective parameters; a possibility to get negative values of the effective parameters; ultra-low values of the refractive index. Throughout the thesis, we consider the initial plane electromagnetic wave that is normally incident to the flat boundaries of the chain. The wave has the magnetic induction vector parallel to the axes of rods while the electric intensity vector is perpendicular to the ones. The above effective complex dielectric and magnetic constants have been obtained for the first time by author of the thesis on base of extension of the case of 2-D infinite metamaterial medium to a slab metamaterial scatterer under consideration through the implementation of the Effective Medium Theory (EMT) in appropriate frequency range. The expressions of the appropriate effective constants for the infinite medium are obtained by other authors. These expressions of the effective constants obtained in this thesis takes in account multipole effects for the case of composite with a very small value of the rods volume fraction while dipole effects are taken in account for the case of large volume fraction values. The accuracy of obtained mathematical models was always benchmarked through a comparison with numerical calculations obtained via the implementations of Finite-Domain Time-Difference (FDTD) method for calculating S-parameters of a metamaterial structure under considerations. S-parameters were used to calculate the effective constants by means of the using Nicolson-Ross approach. All of the numerical experiments presented in the thesis have been carried out with the help of the free Meep FDTD software package while analytical modeling has been done using MatLab software. In this thesis, an improved broadband method for determining complex effective refractive index, dielectric and magnetic constants of an arbitrary passive metamaterial has been proposed. Evaluation of the effective parameters is realized using the reflection-transmission S-parameters obtained by simulation or experimental measurements and analytically evaluated interface reflection coefficient of the slab. In consideration of practical party of this thesis, the obtained qualitative and quantitative results in this thesis have allowed to formulating some properties of two component slab metamaterial structures as arrays of metal rods/wires periodically immersed in a dielectric isotropic matrix: 1. The effective electromagnetic properties of infinite 2-D array of copper cylinders immersed in metal-dielectric matrix in the GHz frequencies shows the existence of the enhancement of effective dielectric constant and low absorption in the microwave frequencies. 2. The obtained analytical models of the composite in the thesis gives a good qualitative but a weak quantitative correlation with results of numerical simulations in the case if cylinders touch each other. 3. The above analytical models of infinite metamaterial medium quantitatively describes well the slabs embedded with the above metamaterial medium if there is some relation between the width of slabs and the dimension of unit cell of the metamaterial medium for appropriate frequency range. 4. The considered artificial material medium can be used to increase the directivity of patch antenna and to obtain ULI structures in the GHz frequency range and to design a new type of waveguides. 5. The obtained mathematical models cannot reveal negative values of the effective dielectric and/or magnetic constants (their real parts) in the GHz frequency range. The main theoretical results of this thesis can be presented by two theoretical methods of characterizing of any 2-D slab metamaterial structures in the microwave frequencies via EMT approach: 1. Non-destructive broadband method for the evaluation of the effective complex dielectric and magnetic parameters of 2-D slab metamaterials. 2. The analytical and numerical optimization method for separating a slab metamaterial into its elementary sub-slabs of the order of the unit cell dimension of the slab. It is important to mention that the above methods have been designed irrespective to the shape of inclusion in the unit cell. Moreover these methods allow to easy evaluating the optical and transport properties of slab metamaterial structures including magnetic ones trough using the relation for the total reflection and transmit ion coefficients and the above constants of single layer. The obtained results in this thesis are in a good quantitative and qualitative agreement with the results of experimental research carried out earlier by one of the supervisors. Moreover these results can be used for creating the course of laboratory works with the using of personal computers for students of Engineer and Sciences directions (Industrial Mathematics, Theoretical Physics, Electrical & Electronic Engineering, Material Science) to study the optical and transport properties of slab metamaterial structures in the microwave frequency range.