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

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Dr Bhanu K Bhakhri

"The Journal of Clinical and Diagnostic Research (JCDR) has been in operation since almost a decade. It has contributed a huge number of peer reviewed articles, across a spectrum of medical disciplines, to the medical literature.
Its wide based indexing and open access publications attracts many authors as well as readers
For authors, the manuscripts can be uploaded online through an easily navigable portal, on other hand, reviewers appreciate the systematic handling of all manuscripts. The way JCDR has emerged as an effective medium for publishing wide array of observations in Indian context, I wish the editorial team success in their endeavour"

Dr Bhanu K Bhakhri
Faculty, Pediatric Medicine
Super Speciality Paediatric Hospital and Post Graduate Teaching Institute, Noida
On Sep 2018

Dr Mohan Z Mani

"Thank you very much for having published my article in record time.I would like to compliment you and your entire staff for your promptness, courtesy, and willingness to be customer friendly, which is quite unusual.I was given your reference by a colleague in pathology,and was able to directly phone your editorial office for clarifications.I would particularly like to thank the publication managers and the Assistant Editor who were following up my article. I would also like to thank you for adjusting the money I paid initially into payment for my modified article,and refunding the balance.
I wish all success to your journal and look forward to sending you any suitable similar article in future"

Dr Mohan Z Mani,
Professor & Head,
Department of Dematolgy,
Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018

Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."

Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018

Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."

Dr Kalyani R
Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018

Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
As an experienced dentist and an academician, I proudly recommend this journal to the dental fraternity as a good quality open access platform for rapid communication of their cutting-edge research progress and discovery.
I wish JCDR a great success and I hope that journal will soar higher with the passing time."

Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
On Sep 2018

Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."

Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata

Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
On Aug 2018

Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".

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

Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".

Dr. Mamta Gupta
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018

Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.

Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."

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)
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.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."

Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
On April 2011

Important Notice

Experimental Research
Year : 2012 | Month : August | Volume : 6 | Issue : 6 | Page : 1105 - 1108

Estimation of the Infarct Size on Occlusion of the Middle Cerebral Artery in Primates

K.S. Satheesha, M.S. Somesh, Shakunthala R Pai, Girish V Patil

1. Assistant Professor, Department of Anatomy, SIMS & RC, Mangalore, India. 2. Assistant Professor, Department of Anatomy, SIMS & RC, Mangalore, India. 3. Professor & Head, Department of Anatomy, SIMS & RC, Mangalore, India. 4. Associate Professor, Department of Anatomy, SIMS & RC, Mangalore, India.

Correspondence Address :
Dr. Somesh M.S. Assistant Professor, Department of Anatomy, Srinivas Institute of Medical Sciences & Research Centre (SIMS &RC), Mukka, Mangalore - 574146, India. Phone: 91 – 0824 – 2425966, 09686924032 E-mail:


Introduction: The model of single artery occlusion in subhuman primates was studied, to verify whether there is any correlation between micro-circulatory perfusion impairment and area of in¬farction after occlusion of middle cerebral artery and to find out whether there is any definite “reperfusion window’ which could be used effectively to prevent or reduce the area of infarction. Materials and Methods: For the present study 24 healthy adult monkeys of either sex were procured and the Middle cerebral artery (MCA) was occluded and reperfused for given set of time periods and the infarction size of the brain was determined. Results: The percentage of the infarction size after occluding the blood vessel increased considerably, as the time interval prolonged. After the reperfusion of the arteries for shorter inter¬vals like 30 min, 4 hours and 12 hours, there was a significant decrease in the infarct size, whereas there was no effect on the infarct size after 12 hours. Conclusion: In the regions of focal cerebral ischaemia, the ad¬verse effect of focal ischemia may be minimized if, in the acute phase, functional microcirculation is increased by some thera¬peutic intervention aimed at augmenting blood flow through the ischemic region.


Cerebral Ischaemia, Occlusion, Reperfusion, Infarct.

How to cite this article :

K.S. Satheesha, M.S. Somesh, Shakunthala R Pai, Girish V Patil. ESTIMATION OF THE INFARCT SIZE ON OCCLUSION OF THE MIDDLE CEREBRAL ARTERY IN PRIMATES. Journal of Clinical and Diagnostic Research [serial online] 2012 August [cited: 2018 Oct 20 ]; 6:1105-1108. Available from

Cerebral ischaemia, usually of the Middle Cerebral Artery (MCA) territory, is one of the common causes of cerebrovascular dis¬eases. The human syndrome of stroke consists of the abrupt development of a focal neurological deficit whose origin can be traced to either the occlusion of a cerebral vessel (usually arterial), or to the spontaneous rupture of an intracranial artery, with a consequent haemorrhage in the brain parenchyma or in the subarachnoid space (1). The retrospective analyses of a large group of stroke patients have revealed that a vast majority were afflicted with one of the 3 anatomical lesions. About 75% of all the stroke affected cases are the clinical expression of brain in¬farctions and large, usually single, brain haemorrhages constitute the background for about 11% of the strokes and primary, non-traumatic, subarachnoid haemorrhages make up almost 5% of all the strokes (2),(3),(4). Brain infarction is a localized destructive lesion which owes to the occlusion of a brain vessel, usually an arterial one. On the basis of long term, repeated physiological and clinical observations, the model of a single artery occlusion in subhuman primates has been found to be the closest to an ideal model of ischaemic stroke (5). The clinical and the morphologic features of this model are very similar to those of massive ischaemic stroke (brain infarction) in human cerebral hemispheres (6). The transorbital surgical approach to the initial segment of the MCA was first described in the 1970s (7). The reliability of this method to induce either an infraction or tissue abnormalities that evolved into an infraction has been confirmed in many laborato¬ries around the world (8),(9). This method allows reperfusion of the ischaemic territory, an important pre-requisite in the development of experimental methods of transient ischaemic attacks. An ex¬tensive injury to the neurons precedes the development of micro-vasculature obstruction and neuronal destruction which is more widespread than impaired perfusion (10). Different experimental approaches have shown that under physiological conditions, all the brain capillaries are perfused continuously with the plasma (11). Under the ischaemic conditions, the swelling of the astrocytes and the capillary endothelial cells or a destruction of the micro-vascular basal lamina occurs during the first hours of the ischaemia, resulting in capillaries with narrowed or total occluded lumens (12). This results in a decrease in the capillary perfusion velocities and later, in the non-perfusion through the capillaries. The aim of the present study was to verify whether there was any correlation between the micro-circulatory perfusion impair¬ment and the area of infarction after the occlusion of the middle cerebral artery and to find out whether there was any definite “reperfusion window’ which could be used effectively to prevent or to reduce the area of the infarction.

Materials and Methods
For the present study, 24 healthy adult monkeys (Macaca radi¬ate) of either sex were procured from non- forest areas with prior permission from the forest Department, Government of Karnataka, India. They were maintained in the animal house of Kasturba Medi¬cal College, Mangalore, Manipal University, India, till the experiment was done and they were maintained accordingly after obtaining the ethical clearance for the present study. Anaesthesia was induced by giving intravenous injections of Nembutal (pentobarbital) (25mg/ml of distilled water for the injection/kg) and it was maintained in a supplementary, minimum dose as and when it was necessary, through the peripheral leg vein, by using a butterfly needle initially.

Material and Methods

For the present study, 24 healthy adult monkeys (Macaca radi¬ate) of either sex were procured from non- forest areas with prior permission from the forest Department, Government of Karnataka, India. They were maintained in the animal house of Kasturba Medi¬cal College, Mangalore, Manipal University, India, till the experiment was done and they were maintained accordingly after obtaining the ethical clearance for the present study. Anaesthesia was induced by giving intravenous injections of Nembutal (pentobarbital) (25mg/ml of distilled water for the injection/kg) and it was maintained in a supplementary, minimum dose as and when it was necessary, through the peripheral leg vein, by using a butterfly needle initially.

After the scarification, the disposals of the animals were carried out through an environment friendly incinerator which was maintained by our university hospital.

a) Surgical Methods:
Under aseptic conditions, the femoral vein and artery were exposed and they were canalized with a 18 G polyethylene tube for the intermittent infusion of the anaesthesia (whenever it was required) and for the blood pressure recording. The anaesthetized animal’s head was fixed firmly in a stereotaxic head holder. The right eye was enucleated and the orbital contents were removed to expose the bony orbit through a transorbital approach. With the help of an operating microscope, the lateral wall of the orbit was widened by using a high speed electric dental drill. The dura matter was cut and the animals were divided in to 3 groups as control, permanent occlusion and temporary occlusion, with 8 animals in each group.

Group I - Control Animals

Group II - Permanent occlusion

Group III - Temporary occlusion

In group I, the MCA was freed from the arachnoid matter and it was exposed; in group II, the MCA was occluded with an aneurysm clip permanently for the given time periods; and in group III, the MCA was temporarily occluded and it was removed after different time periods of the occlusion. The reflow was established and 1½ hours, 4 hours, 12 hours and 24 hours were chosen as the time periods of the study. The dural opening and the orbit were sealed with dental cement to prevent a CSF leak. The whole procedure was carried out under aseptic conditions and the animals were given an antibiotic coverage of 250mg bd penicillin. A periodical blood gas analysis was done and the Blood Pressure (BP) was recorded periodically to ensure that the animals were maintained under normal physiological conditions. The body temperature was monitored with a rectal thermometer and it was maintained within normal limits by using a heating pad (*).

b) Quantification of the ischaemic area:
At the end of the experimental period, the brains of the animals were quickly removed and they were sliced with improvised slicer at the levels of the optic chiasm, the temporal pole and the mammilary body. The slices were incubated at 37°C for 15 minutes in a 1% solution of TTC in saline. The reaction was terminated by substituting the incubation medium with 10% formalin (13). By this method, the normal area was stained red, while the ischaemic area was left unstained. The ischaemic area was quantified with a locally improvised grid in millimeters (mm2). At the end of the selected time period, for the study, the animals were heparinized and perfused with 10% buffered formal saline, after cutting the superior vena cava. Once the animals were properly formalin fixed, the formalin was washed off with warm saline, followed by a treatment with India ink in gelatin, till the animals turned black (14). After keeping them overnight at 4°C, their brains were removed for the quantification of the perfusion defect. An improvised grid was used for the quantification. The total infarct size was determined by TTC (mm2) in millimeters after the occlusion and the reperfusion of the blood vessel for various time intervals, viz 30 minutes, 4 hours, 12 hours and 24 hours and these values were compared with those of the control groups. The statistical analysis of the data which was obtained was done by using the SPSS software, version 6 and the Student’s t test was applied for the comparison between the groups. A p value of ≤ 0.01 was considered as significant.


(A) Physiological parameters: Various physiological parameters like the rectal temperature, blood pressure, etc were recorded for the primate animals, before and 4 hours after the cerebral occlusion, as has been shown in (Table/Fig 1), as a part of the routine protocol.

(B) The cerebral infarction:
In the brains of the control animals (group I), there was no evidence of an infarction, as the blood supply was not interrupted and the brain of these primates looked relatively healthy (Table/Fig 4). The percentage of the infarction size was determined by TTC (mm2) in millimeters in the permanent occluded group (group II) after occluding the blood vessel for various time intervals, viz 30 minutes, 4 hours, 12 hours and 24 hours. As was expected, the infarct size was found to increase considerably as the time interval was prolonged, as has been shown by the arrows in (Table/Fig 5) (a) and (Table/Fig 5) (b)]. The values of the infarct size were also determined, which were significant when they were compared with those of the control animals for the same given time intervals (Table/Fig 2) (Table/Fig 3).

In the group III animals, after the occlusion of the arteries for 4 hours 12 hours , the reperfusion of the arteries was done for various time intervals, viz 30 minutes, 4 hours, 12 hours and 24 hours respectively and the changes in the infarct size were determined. In the brains with the 4 hrs occlusion and reperfusion for 30 minutes, 4 hours and 12 hours, – the infarct size was found to be decreased (Table/Fig 6)(a)and (Table/Fig 6)(c) and the values were significant as compared to those of the controls, whereas the reperfusion for 24 hours had no beneficial effects (Table/Fig 3). Also, in the occlusion of the arteries for 12 hours with reperfusion for various time intervals, there were no changes in the infarct size (Table/Fig 3).


The ability to determine the location and the extent of the areas of infarction in the cerebrum after ischaemia is essential for the assessment of medical or surgical interventions for the confirma¬tion of the clinical findings and for the evolution of new diagnostic techniques. The traditional histopathological methods which are based on the staining with Hematoxylin and Eosin can accurately and reliably distinguish the infracted from the normal tissues. But the infarction is identified during its microscopic examination by its shrinkage, by the dark staining of the neurons and by the swelling with vacuolization of the perineural and the perivascular glial ele¬ments (15),(10). The shrinkage of the tissue during its preparation 1112for the microscopic examination may affect the estimate of the size of the area of the infarction. Originally, (2),(3),(5)- Triphenyltetrazolium hydrochloride (TTC) was used to test the viability of seeds (16) and since 1958, it has been used as a stain to detect the ischaemic infarction in mammalian tissues (17). While this water soluble salt is not a dye, it is reduced by certain enzymes in the normal tissues to a deep red, fat soluble, light sensitive compound (formazan), that turns the normal tissues into a deep red, thereby clearly delineating the abnormal areas (16).

Depending on the physiological factors that affect the cerebral tissue (15), ultra structural changes may appear after 15 to 30 minutes of the ischaemia (18), as the rate of the cerebral oxygen consumption (CMRO2) is reduced in the immediate post ischae¬mic period, as was demonstrated in the rat brain (15). Staining with an agent such as TTC, that is based on the presence of an intact enzyme electron transport chain, might be expected to de¬lineate the infarction at an earlier stage than the traditional histo¬logic methods. The TTC staining is an excellent research method that can be used to confirm the size and the location of the areas of the infarction, and it may be valuable in determining the pres¬ence and the location of the infarction in autopsy samples.

Local blood flow changes are brought about by the modification of the micro-circulation in physiological and pathological states (19). In focal cerebral ischaemia, a reduction in the blood flow has been well documented in the clinical practice (20),(21). A morpho¬metric study of the functional microvasculature was performed on ischaemic cats (22) and also in asphyxiated (23), hypoxic (24), haemorrhagic (25) and hypercapnic rats (11).

Since it was difficult for the study to delineate only the patent micro-vasculature, the India ink perfusion method was adopted. A transcardiac infusion which was done by using the animal’s own systolic pressure at the beginning of the perfusion, avoided the errors of the technique such as variations in the infusion pres¬sure, size of the cannula, and partial obstruction of the tip of the canula against the wall of the vessel (26). The perfusion mixture was prepared just before its use, to prevent the possible varia¬tions due to the differences in its viscosity.

Also, there is a lot of literature on cerebral ischaemia. In a study, when the focal ischaemia due to the Middle Cerebral Artery Oc¬clusion (MCAO) was induced in halothane anaesthetized hyper¬tensive rats, the most marked initial impairments in the perfusion were observed in the core MCA territory and a secondary perfu¬sion impairment was found to develop over time in the peri-focal region (27). The changes in the interstitial oxygen tension (pO2) in the cerebral ischaemic regions, particularly in the ischaemic core and the peri-focal area, induce a complex series of molecu¬lar pathways which involve the signaling mechanisms, gene tran¬scription, protein formation, etc and free radicals and oxidative stress have been suggested to be involved in each of the steps in the injury cascade (28). Also, cerebral ischaemia, followed by the vessel ischaemia/reperfusion of the MCA was studied and it was noted that it alters the vessel properties of the brain arteries in rats, thus inducing an inflammatory response and excessive generation of the reactive oxygen species, leading to an increase in the wall thickness, the cross-sectional area and the wall/lu¬men, and decreased wall stress, which were prevented by the treatment with CR-6, a vitamin-E analogue, that is an antioxidant and a scavenger of the nitrogen-reactive species (29). Also in an other similar study, the administration of oral CR-6, immediately within 2 hours of ischaemia of the MCA occlusion, resulted in the reduction of the glutathione consumption and the superoxide generation in the ischaemic brain, with a decrease in the infarct volume and an ischaemia induced neurological deficit, thus protecting the brain from reperfusion injury but not against perma¬nent ischaemia (30).


So in conclusion, in the regions of focal cerebral ischaemia, where the blood flow is deficient, as was demonstrated in the present study, the adverse effect of focal ischaemia can be prevented or minimized, if in the acute phase, the functional microcirculation is increased by some therapeutic intervention which is aimed at augmenting the blood flow through the ischaemic region.


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ID: JCDR/2012/4646:2344

Date of Submission: Jun 06, 2012
Date of Peer Review: Jun 28, 2012
Date of Acceptance: Jul 12, 2012
Date of Publishing: Aug 10, 2012

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