Asif iqbal ppt Slides on cancer for defense
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This study investigates the susceptibility of cancer patients to bacterial infections and analyzes blood cell counts and c-reactive protein (CRP) levels. A total of 200 clinical specimens were collected, revealing 110 pathogenic bacterial isolates, with 18 being extended-spectrum beta-lactamase (ESBL) producers. Recommendations include enhanced ESBL screening, antibiotic usage awareness, and the need for further research on the microbiological and immunological aspects of these infections.
![Conti…
• C-Reactive protein CRP is a significant acute phase complementary system associated protein.
• Which is present in normal serum of humans and produced by macrophage which release cytokines when
bacteria is engulf by macrophage.
• CRP can be precipitated by C-reactive substances which occur in numerous microorganisms – e.g.
Streptococcus pneumoniae (the basis of the C-reactive protein diagnostic test) and Aspergillus fumigatus
[Binding of CRP to C-reactive substance of Aspergillus fumigatus].
• Changes in CRP level may help to differentiate between bacterial and viral infections.
• A rise in the serum level of CRP is more likely to reflect a bacterial infection because viral infections typically
do not produce such a change.
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Asif iqbal ppt Slides on cancer for defense
- 1. 1
- 2. Comparative Analysis of Complete Blood Count in Cancer and Non- Cancer Patients Fallowed by Antibiogram Analysis of Isolated Bacterial Pathogens Department of Microbiology & Biotechnology Abasyn University Peshawar 2 By M.Phil Scholar Asif Iqbal khattak Reg no:9229 Supervised: Dr. Riaz Muhammad Co-supervised: Dr. Robina Wazir
- 4. • Cancer results from a series of molecular events that fundamentally alter the normal properties of cells. • The genes involved in the development of cancer are classified as oncogenes. • Cancer in developing nations is increasing due to polluted environmental conditions and soon to be a key problem as life expectancy rises. • There are 11 million reported cancer cases globally, and more than half (6 million) happen in low and intermediate income countries. • The American Cancer Society (ACS) assessments that this number will rise to 27 million cases by 2050, generally due to increasing global population and aging. • According to WHO 9.7 million new cases are noticed every single year and 6.7 million Individuals dying due to cancer. 4
- 5. Bacterial pathogens involve in cancer patient infections • Cancer patients have more possibility of getting opportunistic infection i.e. tuberculosis, pneumonia, diarrhoea, gonorrhoea, meningitis and many other infections. • Bacteria pathogens involve in various types of cancer patients are (Salmonella, Shigella, Campylobacter sp, Mycobacterium tuberculosis, Escherichia coli, Listeria sp, Actinobacter baumannii, Salmonella sp, Staphylococcus aureus, Vibrio cholera and Yersinia sp). • Because these patients are immune-compromised due to cancer causing abnormalities. • The misuse of antimicrobial agents perform a key role in the development of MDR bacteria especially Extended Spectrum Beta Lactamases (ESBL) producing bacteria like Salmonella, Proteus, Shigella, Escherichia coli. • Antibiotic resistant Bacteria especially ESBL producing pathogen spread rapidly because these pathogens are very small entities it is transfer from one country to another country by travelling of people. • ESBLs producing bacteria are the most abundant in Enterobacteriaceae family, with more than 600 natural variants. 5
- 6. Conti… • In recent times growing evidence indicated that blood leucocytes, i.e. monocytes and lymphocytes which is further developed into macrophage’s are significantly interconnected to the diagnoses and treatment of various types of cancers. • There is a strong communication between leucocytes and microbial infection. Leucocytes play a very critical role to defend against microbial invasion. • Especially neutrophils, monocytes, dendritic cells and lymphocytes. • The platelets also perform an important role in immune system of our body during bacterial and parasitic infections. • The analysis of Complete Blood Count (CBC) is done through two ways. The programmed or automatic system and a manual system. 6
- 7. Conti… • C-Reactive protein CRP is a significant acute phase complementary system associated protein. • Which is present in normal serum of humans and produced by macrophage which release cytokines when bacteria is engulf by macrophage. • CRP can be precipitated by C-reactive substances which occur in numerous microorganisms – e.g. Streptococcus pneumoniae (the basis of the C-reactive protein diagnostic test) and Aspergillus fumigatus [Binding of CRP to C-reactive substance of Aspergillus fumigatus]. • Changes in CRP level may help to differentiate between bacterial and viral infections. • A rise in the serum level of CRP is more likely to reflect a bacterial infection because viral infections typically do not produce such a change. 7
- 8. Aims of the present study were to investigate the susceptibility of various types of cancer patients toward bacterial infections. Furthermore to analyze the number of blood cells and protein contributing in immunity during bacterial infections & malignancies. Objectives of the current study were as follows: I. Identification and antibiotic susceptibility of bacterial pathogens isolated from blood & urine samples of various cancer and non-cancer patients. II. Screening of ESBL producing bacteria among the isolated bacterial pathogens. III. Comparative analysis of Urine, CBC and CRP in cancer and non-cancer patients. The aims and objectives of the present study were: 8
- 10. 10 Collection of patient specimens LRH Peshawar HMC Peshawar Irnum Hospital Peshawar Transported to laboratory Microbiology research lab of AUP and Microbiology lab HMC
- 11. 11 Isolation of pathogenic bacteria from collected clinical specimens Identification Gram staining Biochemical test Coagulase Catalase Oxidase Urease Indole TSI Cell and colony Morphology Microscopy
- 12. 12 Antibiotic susceptibility Disc diffusion assay Blood analysis Urine analysis Physical Chemical Microscopic Sysmex and Cell dyne ruby for CBC analysis ESBL DETECTION C- reactive protein test Smear method
- 13. The List of Antibiotics used in the study. S/No Abbreviation Antibiotics 01 VA Vancomycin 02 CN Gentamycin 03 AMC Amoxillin 04 CAZ Ceftazidime 05 TZP Tazobactam 06 TE Tetracycline 07 CTX Cefotaxime 08 CIP Ciprofloxacin 09 RD Rifampicin 10 DO Doxycycline 11 C Chloramphenicol 12 PB Polymaxin-B 13 P Penicillin 13
- 14. Results 14
- 15. 15 100 100 200 0 50 100 150 200 250 Blood Urine Total Samples Total sample
- 16. Age limit Male samples Female sample Total Samples 00-35 22 19 41 36-45 20 21 41 46-55 23 18 41 56-65 21 17 38 66-75 23 16 39 Total 109 91 200 16
- 17. 17 00-35 36-45 46-55 56-65 66-75 Total 22 20 23 21 23 109 19 21 18 17 16 91 41 41 41 38 39 200 SAMPLE DISTRIBUTION ACCORDING TO AGE AND GENDER Male samples Female sample Total Samples Age limit
- 18. 18 200 90 110 51 59 30 80 0 20 40 60 80 100 120 140 160 180 200 220 Total Samples Negative Samples Positive Bacterial Isolates Bacterial Isolates in Cancer Samples Bacterial Isolates in Non-Cancer Samples Bacteria Isolates from Blood Samples Bacteria Isolates from Urine Samples Positive and Negative samples
- 19. Percentage distribution of various bacterial pathogens identified in both cancer and non-cancer sample. 19 S No. Bacteria Isolates Count in Cancer Patients Count in Non- cancer Patients TOTAL Count Percentage (%) 01 E. coli 09 07 16 13.33% 02 S. aureus 06 08 14 11.66% 03 P. aeruginosa 06 08 14 11.66% 04 Salmonella 05 07 12 10% 05 bacillus spp. 06 05 11 9.16% 06 Enterobacter spp. 06 04 10 8.33% 07 Mycobacterium Spp. 06 03 09 7.5% 08 S. pyogene 05 04 09 7.5% 09 H. pylori 08 00 08 6.66% 10 Klebsiella spp. 04 03 07 5.83% 11 S. epidermidis 05 00 05 4.16% 12 Shigella 03 02 05 4.16% Total 69 51 120
- 20. 20 9 6 6 5 6 6 6 5 8 4 5 3 69 7 8 8 7 5 4 3 4 0 3 0 2 51 16 14 14 12 11 10 9 9 8 7 5 5 120 1 2 3 4 5 6 7 8 9 10 11 12 Total Bacteria Isolated of cancer patients bacteria isolated from non cancer patients COUNT IN CANCER PATIENTS COUNT IN NON-CANCER PATIENTS Number of Isolates
- 21. 21 0 10 20 30 40 50 60 70 80 90 100 Count VA % R VA % S CN% R CN% S CAZ% R CAZ% S TZP% R TZP% S
- 22. 22 0 10 20 30 40 50 60 70 80 90 Count TE % R TE % S AMC % R AMC % S CTX % R CTX % S CIP % R CIP % S
- 23. 23 0 10 20 30 40 50 60 70 80 90 100 RD % R RD % S DO % R DO % S C % R C % S PB % R PB % S Pen % R Pen % S
- 24. 24 Name of Bacteria Isolated Total number of bacterial Isolates Total ESBL Positive Isolates ESBL Isolates In Cancer Patients ESBL Isolates In Non-Cancer Patients E. coli 16 6 4 2 P. aeruginosa 14 4 2 2 Enterobacter spp. 10 4 3 1 Klebsiella spp. 07 3 1 2 Shigella 05 1 0 1 Total 52 18 10 8
- 25. 25 16 14 10 7 5 52 6 4 4 3 1 18 4 2 3 1 0 10 2 2 1 2 1 8 0 5 10 15 20 25 30 35 40 45 50 55 60 Total number of bacterial Isolates Total ESBL Positive Isolates ESBL Isolates In Cancer Patients ESBL Isolates In Non-Cancer Patients
- 26. ESBL producing E. coli strain. Confirmed with Amoxillin + clavulanate, cefotaxime (a) versusceftazidime alone (b).Figure 2. Inhibition zone differences produced by test cefotaxime alone and in combination with clavulanate. Extended-spectrum beta-lactamase producing E. coli strain. Confirmed with cefotaxime- clavulanate (c) versus cefotaxime alone (d). 26
- 27. C-Reactive Protein analysis for cancer patients Total samples Lowest (0.92 mg/L) Second (0.92-1.93 mg/L) Third (1.94-3.69 mg/L) Highest (>3.69 mg/L) 50 21 08 11 10 27
- 28. 28 50 21 8 11 10 0 10 20 30 40 50 60 Total samples Lowest (0.92 mg/L) Second (0.92-1.93 mg/L) Third (1.94-3.69 mg/L) Highest (>3.69 mg/L) C-Reactive Protein analysis of cancer samples
- 29. C-Reactive Protein analysis for non- cancer patients Total samples Lowest (0.92 mg/L) Second (0.92- 1.93 mg/L) Third (1.94-3.69 mg/L) Highest (>3.69 mg/L) 50 09 11 25 03 29
- 30. 30 50 9 11 25 3 0 5 10 15 20 25 30 35 40 45 50 55 Total samples Lowest (0.92 mg/L) Second (0.92-1.93 mg/L) Third (1.94-3.69 mg/L) Highest (>3.69 mg/L) C-Reactive Protein analysis for non-cancer samples
- 31. 31 0 5 10 15 20 25 30 35 40 45 50 55 60 50 21 23 6 24 12 14 21 20 29 43 Urine analysis of cancer samples
- 32. 32 0 5 10 15 20 25 30 35 40 45 50 50 19 20 9 20 16 14 26 24 20 39 Urine analysis of non-cancer samples
- 33. Complete Blood Count of cancer patients for the evolution of infections. Total sample Lowest WBC (2000- 4000) Highest WBC (5000- 10000) Numerous WBC Lowest HGB (05-10) Highest HGB (10-15) HGB (15-20) Lowest Platelets Below normal Normal Platelets 150,000 -450,000 Highest Platelets Above Normal 50 19 20 11 10 25 15 11 09 30 33
- 34. 34
- 35. 35 0 5 10 15 20 25 30 35 40 45 50 Normal (05-10) (10-15) (15-20) Below normal Above Total sample Lowest WBC (2000-6000) Highest WBC (6000- 10000) Numerous WBC Lowest HGB Highest HGB HGB Lowest Platelets Normal Platelets 150,000 - 450,000 Highest Platelets 50 19 20 11 10 25 15 11 9 30 Blood Analysis Of Cancer Patients
- 36. Complete blood count of Non-cancer patients for the evolution of infections. Total sample Lowest WBC (2000- 6000) Highest WBC (6000- 10000) Numerou s WBC Lowest HGB (05-10) HGB (10-15) Highest HGB (15-20) Lowest Platelets Below normal Normal Platelets 150,000 -450,000 Highest Platelets Above Normal 50 15 15 20 06 25 19 15 15 20 36
- 37. 37 0 5 10 15 20 25 30 35 40 45 50 Normal (05-10) (10-15) (15-20) Below normal Above Total sample Lowest WBC (2000-6000) Highest WBC (6000-10000) Numerous WBC Lowest HGB HGB Highest HGB Lowest Platelets Normal Platelets 150,000 - 450,000 Highest Platelets 50 15 15 20 6 25 19 15 15 20 Blood analysis of Non-cancer patients
- 38. Conclusion 38
- 39. • Clinical specimens (200) were collected in which 110 (55 %) pathogenic bacteria were isolated out of which 18 (9.36%) were ESBL producing bacteria. • The isolated cancer bacteria are E. coli, S. aureus, P. aeruginosa, Salmonella, bacillus spp, Enterobacter spp, Mycobacterium Spp, S. pyogene, H. pylori, Klebsiella spp. S. epidermidis and Shigella. • The isolated non-cancer bacteria are E. coli, S. aureus, P. aeruginosa, Salmonella, bacillus spp, Enterobacter spp, Mycobacterium Spp, S. pyogene, Klebsiella spp. S. epidermidis and Shigella. 39
- 40. Conti… • The ESBL producing cancer bacteria are E. coli, P. aeruginosa, Enterobacter spp. and Klebsiella spp . • The ESBL producing non-cancer bacteria are E. coli, P. aeruginosa, Enterobacter spp. Klebsiella spp and Shigella. • In total 50 cancer patients blood sample CRP were analysed that (42%) patients have the Lowest (0.92 mg/L), (19%) was (0.92-1.93 mg/L) (22%) was (1.94-3.69 mg/L) and (20%) patients were on the Highest (>3.69 mg/L) risk. • In 50 non-cancer blood sample the CRP level are (18%) patients were (0.92 mg/L), (22%) patients were (0.92-1.93 mg/L), (50%) were patients (1.94-3.69 mg/L) and (06%) patients were Highest (>3.69 mg/L) risk. 40
- 41. Conti… • The blood analysis were performed for cancer patients in which 60 % patient were high level of WBC and non-cancer patients were 56 % were High level of WBC. • Total 100 urine samples is also microscopically studied for presence of bacteria in cancer which is 86% and 78% in non-cancer samples. • Maximum resistivity (91%) were showed by penicillin and maximum sensitivity (78%) were showed by rifampicin against isolated bacterial pathogens. 41
- 43. • Continuous ESBL screening and supervision are necessary at hospital settings to observe and develop approaches for monitoring and controlling. • Experimental treatment must be unfavourable which the main cause of ESBL development in bacteria. • Community awareness is required for use and mismanagement of antibiotics. • Sale of antimicrobial drugs without prescription must be inhibited to regulate resistance problem. • Alternate approaches must be hopeful to find out novel sources of antibacterial for regulate of bacterial resistance on the way to existing obtainable antibiotics. 43
- 44. Conti… • More research studies are required to study cancer and non- cancerous patients on microbiological, molecular and immunological level. • Advance Studies are required to correlate CRP level in the serum of cancer and bacterial infected patients. • Which will facilitate the physicians to use a diagnostic procedure for diagnosis of cancer and bacterial infected indirectly. • It require long term research, commitment and budget availability. 44
- 45. Dr. Riaz Muhammad (supervisor) Dr. Robina Wazir (Co-Supervisor) Mr shahid Ali khan (Microbiology Section Head) Mr. Shabeer Ahmad (Head of Department) Dr. Muhammad Salman Dr. Muhammad Rizwan Dr. Hayat Khan Mr. Abdur Rehman Mr. Ghadir Ali Mr. Saif Ullah Mr. Khalid Amin Madam Sidra Mr. Umair Ahmad Mr. Jamshaid Khan Mr. Arsalan All colleagues and friends 45 Acknowledgement
- 46. Thank you 46
Editor's Notes
- #3: My name is asif Iqbal my research superviser name is dr raiz my research topic name is ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
- #5: Cancer is a group of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. he genes that code for the positive cell cycle regulators are called proto-oncogenes. Proto-oncogenes are normal genes that, when mutated in certain ways, become oncogenes: genes that cause a cell to become cancerous. There are several ways by which a proto-oncogene can be converted into an oncogene.
- #6: non-specpic doage of antibiotics and for proYou can spread an ESBL infection simply by touching someone or leaving bacteria on a surface that someone else touches. This can include: shaking hands breathing on someone handling an object that is then handled by someone else per duration also a cause of esbl ,,,,, The Enterobacteriaceae are a large family of Gram-negative bacteria that includes, along with many harmless symbionts, many of the more familiar pathogens, such as Salmonella, Escherichia coli, Yersinia pestis, Klebsiella, and Shigella.
- #7: Monocytes are the largest type of leukocyte and can differentiate into macrophages and myeloid lineage dendritic cells. part of innate immune system monocytes A type of immune cell that has granules (small particles) with enzymes that are released during infections, allergic reactions, and asthma. Neutrophils, eosinophils, and basophils are granulocytes. A granulocyte is a type of white blood cell. Hematopoiesis The formation of new blood cells. Dendritic cells (DCs) are antigen-presenting cells
- #8: CRP is produced in the liver and its level is measured by testing the blood. CRP is classified as an acute phase reactant, which means that its levels will rise in response to inflammation. Complement is a system of plasma proteins that can be activated directly by pathogens or indirectly by pathogen-bound antibody, leading to a cascade of reactions that occurs on the surface of pathogens and generates active components with various effector functions. any of a number of substances, such as interferon, interleukin,
- #10: The study will be carried out in Microbiology Research Laboratory (MRL), Abasyn University Peshawar and in Hayatabad Medical Complex (HMC) main Microbiology and Hematology laboratory from March 2017 to October 2017. In This study we will focus on the analysis of CBC between cancerous and non-cancerous patient’s and identification, antibiotic susceptibility of bacterial pathogens isolated from urine and blood sample of cancer patients of Peshawar region Khyber Pakhtunkhwa (KPK).
- #11: Nutrient agar, MacConkey agar, Salmonella, Shigella Agar (SSA), and SSA will be used for identification of salmonella and Shigella. Eosin Methylene Blue (EMB), MacConkey agar is a selective and differential media so it will be used for the growth of Gram negative bacteria.
- #12: Gram-negative produce pink color. , gram-positive produce purple…
- #22: green,,,, light blue ,,,,,,, orange ,,red,,, gray….dark brown,,,,,light blue…. Dark blue,,,,purple VA Vancomycin, CN Gentamycin, CAZ Ceftazidime, TZP Tazobactam, more resistant antibiotic is Tazobactam and less potent antibic is gentamycin
- #23: Dark Red …. Red,,,0range,,,,yellow,,, light green,,,,,, green,,,, light blue,,,, white dark,,,,purple TE Tetracycline, AMC Amoxillin, CTX Cefotaxime, CIP Ciprofloxacin, more resistant antibiotic are Ciprofloxacin and more sensitive antibiotic are Amoxillin
- #24: Green,,,orange,,, dark white,,, dark,,, blue,,,green,,, red,,,,dark red,,, light orange,,,purple RD Rifampicin, DO Doxycycline, C Chloramphenicol, PB Polymaxin-B, P Penicillin., most resistant antibiotic are pencillion and less resistant antibiotic
- #25: of clavulanic acid to cefotaxime and ceftazidime A 0.5 MacFarland’s suspension of every single isolate will be spread on a Muller – Hinton agar (MHA) plate ceftazidime (30 μg) ceftazidime and clavulanic acid (30 μg/ 10 μg) discs will be positioned aseptically on the agar plate.
- #26: McFarland standards were made by mixing specified amounts of barium chloride and sulfuric acid together.
- #28: Normal level of is between 1.0 mg/L and 3.0 mg/L. Your CRP levels should be normal if you don't have any infections or chronic inflammatory illnesses such as RA, Crohn's disease, or lupus. C-reactive protein is measured in milligrams of CRP per liter of blood (mg/L). Normal CRP levels are below 3.0 mg/dL.
- #29: McFarland standards were made by mixing specified amounts of barium chloride and sulfuric acid together.
- #30: . The cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF) are widely reported to induce synthesis of CRP by hepatocytes both in vitro and in vivo.
- #34: The normal number of WBCs in the blood is 4,500 to 11,000 WBC per microliter (4.5 to 11.0 × 109/L). Normal value of wbc is 4000 to 11000 hb14 – 18, For men, 13.5 to 17.5 grams per deciliter. For women, 12.0 to 15.5 grams per deciliter. Pletaltes normal quantity are 150,00 4500 150,000 to 400,000 per microliter