Cloning and chloroplast-targeted expression studies of insect-resistant gene with ricin fusion-gene under chloroplast transit peptide in cotton

Background: Transgenic plants inhabiting single Bt gene are prone to develop insect resistance and this resistance has been reported in case of some important yield-devastating insect larvae of commercial crops, such as cotton and rice. Therefore, it has become essential to adapt new strategies to overcome the problem of insect resistance and these new strategies should be sophisticated enough to target such resistant larvae in broad spectrum. Among these, plants may be transformed with Bt gene tagged with some fusion-protein gene that possesses lectin-binding capability to boost the binding sites for crystal protein gene within insect mid-gut in order to overcome any chances of insect tolerance against Bt toxin. Enhanced chloroplast-targeted Bt gene expression can also help in the reduction of insect resistance. Results: In the present investigation, a combined effect of both these strategies was successfully used in cotton (G. hirsutum). For this purpose, plant expression vector pKian-1 was created, after a series of cloning steps, carrying Cry1Ac gene ligated with chloroplast transit peptide towards N-terminal and Ricin B-Chain towards C-terminal, generating TP-Cry1Ac-RB construct. Conclusions: Efficacy of pKian-1 plasmid vector was confirmed by in-planta Agrobacterium-mediated leaf GUS assay in tobacco. Cotton (G. hirsutum) local variety MNH-786 was transformed with pKian-1 and the stable integration of TP-Cry1Ac-RB construct in putative transgenic plants was confirmed by PCR; while fusion-protein expression in cytosol as well as chloroplast was substantiated by Western blot analysis. Whereas, confocal microscopy of leaf-sections of transgenic plants exposed that hybrid-Bt protein was expressing inside chloroplasts.

Various modes of fatalities and pattern of fatal injuries in the field

To determine various modes of fatalities and pattern of fatal injuries in the field. Descriptive study. The study was carried out at the Combined Military Hospital Bannu, the base hospital of Waziristan FATA from January 2005 to October 2009. All the soldiers brought in dead from operational area to CMH Bannu were included in the study. After entry of their personal data in admission and discharge book the detailed examination was carried out. Only percentages are presented for confidentiality purpose. However details may be provided by the author on requests routed through the authorities concerned. Gunshot wounds [33.51%], bomb blast [30.16%], mine blast [18.99%] and rocket blast [7.82%] were the common modes of fatality. Multiple injuries [two or more than two] were found on 63%, head injuries on 18%, chest injuries on 13% and abdominal injuries on 6% of all the fatal cases. Maximum numbers of fatal cases were received in the year 2005 [37.65%].Most common mode of fatal injuries was gunshot wound followed by bomb blast and mine blast. Multiple injuries were the most common fatal injuries followed by head and chest injuries. There is a need for preparedness at all levels to reduce the mortality.

Pattern of distribution of metastatic lesions within skeleton in patients with breast carcinoma of Faisalabad and its vicinity

Evaluation of pattern of distribution of skeletal metastases in patients with breast carcinoma. by using [99m]Tc MDP Skeletal Scintigraphy. Retrospective Analysis. Study was conducted at Punjab Institute of Nuclear Medicine [PINUM], Faisalabad Pakistan from January 2006 to June 2009. A retrospective study was performed on 465 consecutive patients having CA Breast irrespective of preoperative/postoperative status, presenting for bone scan at Punjab Institute of Nuclear Medicine. Whole body bone scan was performed, along with additional spot views, where needed. The images were interpreted for metastatic deposits by three independent observers. Where needed, plain X-ray was performed to correlate in favor of benign pathology. Out of 465 patients [453 female and 12 Male], 286 [62%] patients were having either normal bone scan or some benign pathology as correlated with plain x-ray of the suspected area. 179 [38%] patients were positive for metastases within skeleton. Among these, 61.4% patients were having multiple, 14% with two, while 24.6% were diagnosed as having solitary lesions within the skeleton. In patients having multiple skeletal lesions, highest number was noted in spine [84.5%- most common in thoracolumbar], followed by ribs [55.5%], pelvis [37.3%---most frequent in iliac bone], skull [32%], scapula [27.3%], sternum [26.4%], femur [19.1%], humerus [14.5%], clavicle [3.6%] and tibia [0.9%]. In humerus and femur, most lesions were located in their upper ends, followed by shafts. Right side of skeleton was more frequently involved than the left. In patients having lesions in ribs and pelvis, most of the lesions were bilateral. Patients with two lesions showed maximum number of lesions in rib cage [44%], followed by spine [32%-all being in thoracolumbar region], scapula [24%], pelvis and scapula [24% in each], sternum [16%], skull [8%], humerous and femur [4% in each]. In patients having solitary focal lesion, highest number was noted in spine [45.5%-most common in thoracolumbar spine], followed by ribs [22.7%], sternum and pelvis [13.6% each], and skull [6.8%]. No lesion was noted in appendicular skeleton. Multiple skeletal metastases are much more common than solitary or two lesions in cases of known breast carcinoma. Axial skeleton including skull is most commonly involved. Spine [especially thoracolumbar] is the most frequent site of involvement followed by rib cage, pelvis and then skull. In peripheral bones, most frequent site is the upper end of femur followed by humerus

Gated spect myocardial perfusion scintigraphy for identifying septal perfusion artifacts in left bundle branch block

To determine the role of gated Single Photon Emission Computed Tomography [SPECT] for accurate assessment of myocardial perfusion scintigraphy [MPS] of patients with left bundle branch block [LBBB]. Analytical study. Punjab Institute of Nuclear Medicine [PINUM], Faisalabad, Pakistan, from June 2002 to April 2003. MPS data of patients with LBBB was analyzed. Resting gated SPECT MPS was performed after an injection of 740 MBq 99mTc-MIBI in 10 normal and 25 subjects with LBBB [with low probability of coronary artery disease]. Visual and quantitative analyses were done on non-gated [NG], end diastolic [ED], end systolic [ES] images. Calculations included septal to lateral wall ratio [SLR], myocardial thickening [MT=% increase in counts during systole] at end systolic phase and myocardial thickening at peak level [% peak MT]. Septal hypoperfusion was noted in 19 [76%] patients on NG images and in only 1 [4%] patient on gated SPECT ED images. On NG images of LBBB group, SLR was lower than in controls [0.68 +/- 0.07 vs. 0.87 +/- 0.05, p<0.001]. SLR of LBBB patients approached to that of control group in gated SPECT ED data [0.86 +/- 0.06 vs 0.88 +/- 0.06, p=ns]. Myocardial thickening at ES for septum was markedly lower in LBBB group than in controls [21.83% ' 10.86 vs. 66.32% ' 20.15, p<0.001]. In patients with LBBB, reduced septal thickening results in artifactual septal perfusion defects. Gating the perfusion scintigraphy and reporting perfusion status on end diastolic frames in LBBB patients can eliminate these artifacts

Diagnosis of coronary artery disease by stress Echocardiography and perfusion scintigraphy

This meta-analysis was performed to compare the diagnostic efficacy of stress echocardiography [SE] and Stress perfusion studies [SPS] in detecting coronary artery disease [CAD]. A meta-analysis of peer reviewed articles, published in English language, reporting head-to-head comparison of vasodilator stress echocardiography [VSE] and SPS for the diagnosis of CAD, was performed. Data of 13 studies comprising of 860 patients from 13 different institutions were analyzed. Algorithms were developed to generate raw data from published papers to calculate statistical parameters with confidence intervals and then compare them at specified significance levels. The overall diagnostic accuracy of the two tests was almost similar, 0.77 for VSE vs 0.8 for SPS [p=ns]. SPS gave higher sensitivity, 0.88 vs 0.70 in cumulative data [p<0.0001] while VSE gave higher specificity, 0.90 vs 0.67 [p<0.0001]. Accuracy of VSE with state-of-the-art protocols became even better than SPS [p<0.05]. In hypertensive patients specificity of SPS was markedly deteriorated. Contrary to this, VSE gave higher specificity [0.90 vs 0.40] in this subgroup of patients as well. VSE might become an effective alternative of SPS where scintigraphy techniques are not available or affordable

99mTc MAG3 and 99mTc DTPA; comparison to evaluate renal functions in renal transplant patients

Renal scintigraphy evaluates the renal function non-invasively. The present study was conducted to compare 99mTc MAG3 and 99mTc DTPA in renal transplant patients for the evaluation of renal functions. Materials and 99mTc MAG3 and 99mTc DTPA studies were performed on 33 renal transplant patients and 10 normal subjects, with 1 day interval in between the two studies. Renal transplant patients were categorized as normal, acute tubular necrosis, rejection, urine outflow obstruction and urinary tract infection groups on the basis of routine clinical investigations and preliminary results of scintigraphic studies. Both radiopharmaceuticals, 99mTc MAG3 and 99mTc DTPA were having equal diagnostic efficacy, however image quality with 99mTc MAG3 was superior to that of 99mTc DTPA. Renograms clearly differentiated various renal groups. Correlation between glomerular filtration rate [GFR] and effective renal plasma flow [ERPF] was good [r=0.89 at P<0.00001]. In terms of perfusion index [PI] our results with both agents were similar [r=0.96 at p<0.00001], clearly differentiating all cases of rejection from acute tubular necrosis. PI was raised in rejection cases, while normal in all other groups. In cases of acute tubular necrosis and rejection, 99mTc MAG3 based parenchymal retention index [R-20/3] was markedly raised, compared to that of 99mTc DTPA [p<0.00001]. 99mTc- MAG3 and 99mTc DTPA have equal diagnostic efficacy in renal transplant patients. However visual delineations of the kidneys is better with 99mTc MAG3, especially in cases with compromised renal functions

Detection of severity of coronary stenosis by technetium-99M-MIBI imaging at rest, a quantitative spect study

Our main aim of the study was to evaluate the potential role of resting technetium-99m-MIBI imaging as a detector of degree of severity of coronary stenosis in patients having an episode of acute myocardial infarction six to twelve weeks before. There were 18 patients underwent resting technetium-99m MIBI perfusion imaging. Scintigraphic results obtained in the form of perfusion indices and angiographic results in the form of percentage stenosis. Perfusion index < 0.65 was considered as tested positive and percentage stenosis > 80 percent without the existence of collateral circulation was taken as diseased positive. The sensitivity, specificity, positive predictive value, and a negative predictive value of the overall test is ninety, eighty five, seventy eight, and ninety four percent respectively. In the myocardial segments supplied by severely stenosed coronary arteries, there was significantly higher MIBI uptake in the segments with good collateral circulation as compared with those subtended by a vessel totally occluded with poor collateral circulation [p<0.01]. It is concluded that quantitative analysis of myocardial tomography with technetium99m MIBI at rest can differentiate between myocardial segments supplied by critically stenosed and non-critically stenosed coronary arteries. It also evaluates the existence of collateral circulation