Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




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Aug 2018




Dr. Rajendra Kumar Ghritlaharey

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Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
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Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2007 | Month : December | Volume : 1 | Issue : 6 | Page : 467 - 475 Full Version

Systemic Immunomodulation and Hypocholesteraemia by Dietary Probiotics: A Clinical Study


Published: December 1, 2007 | DOI: https://doi.org/10.7860/JCDR/2007/.155
PAWAN R**, BHATIA A.*

**Research Scholar, Department of Biotechnology, Punjabi University, Patiala-147002

Correspondence Address :
Professor, Department of Biotechnology, Punjabi University, Patiala-147002.Fax: +91-175-2286682, 2283073, e-mail: aruna_bhatia@rediffmail.com.

Abstract

Probiotics intake has been associated with beneficial effects on the hyporcholesteremia and gut immune system. This improves disease resistance, gut infection and diminished risk were documented. In one of our experimental study in mice, probiotics was found to modulate the immune response and at the same time it decreased cholesteremia. The present study was extended to human beings, male healthy volunteers, to find out whether probiotics given in the form of fermented milk product ‘Dahi’ and ‘Lassi’ show similar effect i.e. enhanced immune response and hypocholesterolemia. The immune response was checked before and after feeding of probiotics by employing INOs and NBT reduction tests. In addition the effect of probiotics on hypertension and cholesteremia was also observed. The results show significantly enhanced immune response by probiotic consumption and non significant reduction in cholesterol level, HDL-C in the human subjects and decrease in systolic blood pressure in hypertensive patients by the consumption of ‘Dahi’ and ‘Lassi’ was also observed. It is suggested that probiotic diet therapy can be a safe additive or alternative to existing drug therapy.

Keywords

INOs; Inducible nitric oxide synthase, NBT; Nitroblue Tetrazolium Chloride, systolic blood pressure, immune response HDL-C; High Density lipoprotein cholesterol.

Introduction
The role of probiotics in human health is well documented now. There are reports which reveal that probiotics results in hypocholesteraemia (3),(21),(24),(31), ameliorate the gut immunity (5),(6),(7),(16),(30) and as a result are helpful in modifying diseases like atherosclerosis and gut infection. But the literature lacks the effect of probiotics on systemic immunity as well as mechanism of hypocholesteraemia. In one of our preliminary studies in mice a direct correlation was observed between immunomodulatory and hypocholesteraemic property of probiotics (10).
Not only this, the immune status of the host may modulate many immunological disorders and immune related diseases e.g. diabetes, cholesteraemia etc. The increase in awareness of the consumer about the side effects and cost of allopathic medicines has resurged the interest of the consumer in alternative therapy. The dietary materials are the most accepted alternative therapeutic agents. Hence the present study was designed to see the effect of consumption of probiotics in the form of ‘Lassi’ and ‘Dahi’, a common Punjabi food on systemic immunity and cholesteremia in human subjects. In addition the effect of probiotics consumption on hypertension was also observed.

Material and Methods

DIET: ‘ ‘Dahi’‘ and ‘ ‘Lassi’‘ made by fermenting the low fat milk, containing ‘lactobacillus sps. ‘, ‘Streptococcus sp. ‘ were procured from Verka Milk Plant Ltd., Mohali and checked for proximate composition and viable cell count 2.1 × 108 cfug-1 with routine microbiological methods were given to each subject to be consumed @ ‘ ‘Lassi’ ‘350 ± 50 g/d/subject and ‘ ‘Dahi’‘ 250 g/d/subject, for consecutive 30 days.

GROUPING: For cholesterol level; Total sixty volunteers were divided in two group; Group-Ia (n=30) as control, the volunteers in this group were not recommended any test diet during the period and test Group IIa (n=30) on test diet for 30 days.

For Hypertensive study:Total 20 hypertensive patients attending OPD of Govt. Rajindra Medical College and Hospital and Amar Hospital, Patiala, Punjab, for regular blood pressure check up were divided into two Groups:
Group–Ib (n=10) (control hypertensive), subjects were not on test diet.
Group-IIb (n=10) (test hypertensive) The patients in this group were having hypertensive disease but were not taking any kind of medication. In addition to the normal diet, this group volunteers took ‘Dahi’ ‘and ‘ ‘Lassi’ ‘daily for 30 days as given above. No attempts were made to alter life style pattern of volunteers and they were told to maintain their usual habits as per the advice of the consultant doctor for both anti-cholesterolemic and hypertensive study (13).

Follow up of study:The study design was described to each volunteer. Each Group was instructed to follow diet guidelines as suggested by White et al. (13) and consume ‘Lassi’ and ‘Dahi’ for 30 days. Multiple blood samples were obtained from each Group on the zero day and 30 day of study and assayed for total serum cholesterol, HDL-C, inducible nitric oxide synthase (INOs) activity and Nitroblue tetrazolium chloride (NBT) reduction test (a measure of immune status).

Estimation of Total serum cholesterol:
Total serum cholesterol was estimated by the method of Wybenga et al. (1) using commercial kit (Diagnostic Reagent Kit manufactured by Span Diagnostic Ltd., India) based on reaction of cholesterol hot solution of ferric perchlorate, ethyl acetate and sulphuric acid, which gives lavender color complex which is measured spectrophotometerically at 540 nm.
The concentration of cholesterol in mg/dl of the test samples was calculated as:

Total Serum Cholesterol (mg/dl) = (O. D. of Test (T)/ O. D. of Standard (s)) x 200

HDL cholesterol (HDL-C): HDL-C was determined by one step method of Wybenga et al. (1) using commercial kit, manufactured by Span Diagnostic Ltd., India, Briefly 0.2 ml of serum sample mixed with 0.2 ml of precipitating reagent and based on reactions, lavender color complex which is measured by using UV-VIS spectrophotometer (Shimadzu) at 540 nm.
The concentration of serum HDL cholesterol in mg/dl of the test samples was calculated as:

Serum HDL-C (mg/dl) =( O. D. of Test (T)/ O. D. of Standard (s)) x 50

Determination of INOS (Inducible nitric oxide synthase test) activity: The iNOS activity was measured in vitro in blood lymphocytes (suspended in MEM @ 1 × 106 viable cells/ml) using arginine and Greiss reagent by the method of Stuehr and Marletta (11). Optical density of the citrulline formed was determined spectrophotometerically with a UV-VIS spectrophotometer (Shimazu) at 540 nm against control.
Determination of Nitroblue Terazolium Test reduction (NBT): The NBT reduction test was employed to assess the function of phagocytes as described by Hudson and Hay (25). Briefly, blood leukocytes were exposed to NBT and development of color due to formazon was measured spectrophoto

Results

The results (Table/Fig 1) and (Table/Fig 2) show consumption of probiotic containing diet (Dahi’ and ‘Lassi’) in Test Group-IIa resulted in 10.30% ±1.32 reduction in total serum cholesterol and 3.56% ±0.8 difference in HDL-C after 30 days on test diet. These differences in total serum cholesterol and HDL-C were insignificant statistically by (p<0.05). The effect of probiotics consumption on immune response is shown in (Table/Fig 3) and (Table/Fig 4). Significantly higher level (p<0.05) of INOs activity 29.30% ±0.61 and NBT activity 16.10% ±0.14 in test Group-IIa was observed as compared to the control Group-Ia.

The results of probiotic consumption in hypertensive patients are shown in the fig. 5. There was 6.93% ±2.0 decrease in systolic blood pressure level in test Group-IIb on fermented milk product for 30 days. However, no marked difference in systolic blood pressure level in control Group-Ib was observed (Table/Fig 5), In addition there was enhancement in immune response (Table/Fig 3), (Table/Fig 4)in these subjects as observed by enhanced INOs and NBT activity. The INOs activity and NBT reduction was 13.13% ±0.27 (p>0.05) and 4.56% ±0.0.04 (p>0.05) more respectively in test Group-IIb as compared to the control group-Ib. Our results show, though insignificant, yet an inverse relationship between the immune activity and level of blood pressure.

Discussion

In the present study the effect of probiotic consumption in the form of ‘Dahi’ and ‘lassi’ was studied on immune response, cholesterol level and blood pressure in normal and hypertensive subjects. Results reveal that consumption of these fermented products show hypocholesteremic, immunomodulatory and hypotensive effect. Our observation that probiotic diet can lower the cholesterol level is in agreement with other experimental (3),(31) and clinical studies (14). Ashar et al. (19) reported hypocholesterolemic effect of probiotic diet in humans and showed total cholesterol reduction to an extent of 12-21% by feeding on acidophilus milk. The lactic fermenters of yogurt reduce the levels of total cholesterol, HDL-C and LDL-C, in a well-balanced way by assimilation of cholesterol by bacteria in youghurt (in vitro) (12). Similar findings were earlier observed by us in experimental animals (18). The effect of probiotic intake on cholesterol level on human subjects carried by Fabian and Elmadfa (24) was quite like our study which supported consumption of fermented milk leads to decrease in cholesterolemia. In our study, reduction in total serum cholesterol and small difference in HDL-C contents in the humans fed on diet containing lactic acid bacteria was observed. Similar results in humans were also reported by Keim et al. (21). However, variable effect of different probiotic microorganisms on serum cholesterol has been given in a review by Shah, 2007 (32). In a study performed by Hashimoto et al. (23), a diet containing L. casei TMC 0409 was found to raise the concentration of HDL-C in the blood. Besides, Fukushima and Nakao (22) and Simons et al. 2006 indicated no significant difference in the HDL-C content or cholesterol level corresponding to supplement of probiotics, including Lactobacillus and Streptococcus, in lipid-rich and cholesterol-rich diets.

In our results the higher values of INOs and NBT in test groups than the control groups show stimulation of systemic immunity. This supports the earlier findings that the probiotic diet might modulate and stimulate immune response (2),(4),(7),(8). Fermented low fat milk product ‘Lassi’ and ‘Dahi’ feeding reduced the cholesterol level and increase immune activity in humans. A similar correlation between immune response and cholesteremia has been earlier observed in our previous study (10). Moreover, Matsuzuki and Chin 2000 (26) reported the oral administration of Lactobacillus casei strain Shirota (LcS) has been found to enhance innate immunity by stimulating the activity of splenic NK cells. However Christensen et al. 2006 (27) could not find any effect of probiotic supplemented diet on immune response of young healthy adults, even with high doses of B. animalis ssp. Lactis BB-12 and L. paracasei ssp. paracasei CRL-43. But Gill et al. (28) observed enhancement in cellular immunity in the elderly on B. lactis as probiotic dietary supplement.

In our results the test groups show enhancement in immune response with simultaneous decrease in blood pressure. Our findings corroborate the earlier report by Kawase et al (15) who stated decrease in systolic blood pressure by fermented milk products.
The role of fermented food in lowering of risk for cardiovascular diseases has been shown earlier by Frick et al. (17) and reduction in serum ch

Conclusion

The available literature reveals that cholesterol and hypertension both causes of atherosclerosis can be modified by the intake of probiotic diet. Diet therapy is cost effective to drug therapy. Moreover the unwanted side effects of the drug can also be minimized. Results reveal that dietary intake of fermented milk product ‘Dahi’ and ‘Lassi’ by humans’ results in decrease in total serum cholesterol and systolic blood pressure and on the other hand significantly increase INOs and NBT activity of macrophages, which show the inverse and indirect relationship of cholesterol and blood pressure with immune activity. Our, this study clearly projects the usefulness of the probiotic containing diet as safe alternative immunotherapeutic agent and functional food for the persons suffering from hypercholesterolemia, hypertension and immune disorder. Not only this, the immune system of body further controls many disorders like diabetes, diarrhea, cancer etc. hence probiotics can improve the overall health of the consumer.

References

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Wybenga DR, Pileggi VJ, Dirstine PH, Giorgio JD. Direct manual determination of serum total cholesterol with a single stable reagent. Clin. Chem 1970;16:980-984 .
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Schiffrin EJ, Rochat F, Link-Amster H, Aeschlimann JM, Donnet- Hughes A. Immunomodulation of human blood cells following the ingestion of lactic acid bacteria. J Dairy Sci 1995;78:491–497.
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Lin J, Lin EC, Yu LT, Liu HT, Yang TS, Huang CH. Effect of probiotic supplementation on growth performance, serum cholesterol and triglyceride immune response and fecal bacteria in early weaned piglets. J Agri Ass of china 2002;3(4): 325-336.
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Bujalance C, Moreno E, Jimenez-Valera M, Ruiz-Bravo A. A probiotic strain of Lactobacillus plantarum stimulates lymphocyte responses in immunologically intact and immunocompromised mice. Int J Food Microbiol. 2007 Jan 1;113(1):28-34.
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Dunne C, Shanahan F. Role of probiotics in the treatment of intestinal infections and inflammation. Curr Opin Gastroenterol 2002 Jan;18(1):40-5
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Fedorak RN, Madsen KL. Probiotics and prebiotics in gastrointestinal disorders. Curr Opin Gastroenterol. 2004 Mar;20(2):146-55.
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Meydani SN, Woel-Kyu H. Immunologic effects of yogurt. Am J Clin Nutr 2000;71:861–872.
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Solis-Pereyra B, Lemonnier D. Induction of 29-59 A synthetase activity and interferon in humans by bacteria used in dairy products. Eur Cytokine Netw 1991;2:137–140.
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Kiyoshi T, Takeya M, Sakashita N. Multi functional roles of macrophages in the development and regression of Atherosclerosis in human and experimental animals. Med Elect Microscopy 2004: 35(4): 179-203.
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Ritu Pawan, Bhatia, A. Correlation and modulation of immune response and serum cholesterol level. Punjabi University Patiala: M.Sc. Dissertation 2003.
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Stuehr DJ, Marletta MA. Synthesis of nitrite and nitrate in murine macrophage cell lines. Cancer Res 1987;47:590-594.
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Abdelkader DB. Assimilation (In Vitro) of cholesterol by yogurt bacteria. Ann Agric Environ Med 2006;13:49–53
Tables and Figures
[Table / Fig - 1] [Table / Fig - 2] [Table / Fig - 3] [Table / Fig - 4] [Table / Fig - 5]

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