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Biomedical subjects

M Singh

Publications and source records attributed to M Singh.

At least 613 records · Page 34Linked to original sources
PubMed1994

Potentiation of ricin A immunotoxin by monoclonal antibody targeted monensin containing small unilamellar vesicles.

The carboxylic ionophore monensin could be successfully entrapped in small unilamellar vesicles made by the extruder method. Monensin liposomes of size range 100-150 nm were more potent in potentiating ricin A immunotoxin activity in vitro as compared to monensin liposomes of diameter 500 nm or more. These liposomes were further successfully linked to tumor specific monoclonal antibodies with full retention of immunoreactivity. Monoclonal antibody targeted monensin liposomes were 100 times more potent than monensin liposomes in potentiating the activity of ricin A immunotoxins against various tumor cell lines in vitro.

Antibodies, Monoclonal↗
PubMed1994

A molecular rheostat. Co-operative rev binding to stem I of the rev-response element modulates human immunodeficiency virus type-1 late gene expression.

The complete biologically active human immunodeficiency virus type-1 (HIV-1) rev-response element (RRE) RNA is 351 nucleotides (nt) in length, and includes an extra 58 nt on the 5' end and 59 nt on the 3' end beyond the sites proposed in the original models for the RRE secondary structure. The extra sequences are able to form a duplex structure which extends Stem I. The presence of an elongated Stem I structure in the RRE RNA was confirmed by nuclease mapping experiments. Nuclease protection experiments have shown that rev binds to restricted regions of the RRE, including the high affinity site located at the base of Stem IIb and along the length of the Stem I region. The three large stem-loop structures which protrude from Stem I and Stem IIb (Stems IIc, III+IV and V) remain accessible to nucleases even in the presence of a large excess of protein. Gel-retardation experiments show that the truncations of Stem I reduced the total number of rev molecules that can bind co-operatively and with high affinity to the RRE RNA. To test whether the elongated Stem I structure is required for maximal rev activity, a series of truncations which progressively reduced the length of Stem I was introduced into an HIV-1 derived reporter plasmid. In the presence of rev and a functional RRE, there is an increase in the levels of gag and env mRNA in the cytoplasm and a decrease in levels of tat and rev mRNAs. Each of the truncations in Stem I reduced the rev responses, with the longest truncations producing the greatest losses of activity. The data suggest that the RRE acts as a "molecular rheostat" designed to detect rev levels during the early stages of the HIV growth cycle.

Base Sequence↗
PubMed1994

Methylphosphonate mapping of phosphate contacts critical for RNA recognition by the human immunodeficiency virus tat and rev proteins.

The HIV-1 regulatory proteins tat and rev are both RNA binding proteins which recognize sequences in duplex RNA which are close to structural distortions. Here we identify phosphate contacts which are critical for each binding reaction by use of a new method. Model RNA binding sites are constructed carrying substitutions of individual phosphodiesters by uncharged methylphosphonate derivatives isolated separately as Rp and Sp diastereoisomers and tested for protein binding by competition assays. In the binding of tat to the trans-activation response region (TAR), three phosphates, P21 and P22 which are adjacent to the U-rich bulge and P40 on the opposite strand, are essential and in each case both isomers inhibit binding. Similarly, in the interaction between the HIV-1 rev protein and the rev-responsive element (RRE) both methylphosphonate isomers at P103, P104, P124 and P125 interfere with rev binding. At P106, only the Rp methylphosphonate isomer is impaired in rev binding ability and it is proposed that the Rp oxygen is hydrogen-bonded to an uncharged amino acid or to a main chain hydrogen atom. Synthetic chemistry techniques also provide evidence for the conformations of non-Watson-Crick G106:G129 and G105:A131 base-pairs in the RRE 'bubble' structure upon rev binding. Almost all functional groups on the 5 bulged residues in the bubble have been ruled out as sites of contact with rev but, by contrast, the N7-positions of each G residue in the flanking base-pairs are identified as sites of likely hydrogen-bonding to rev. The results show that both tat and rev recognize the major groove of distorted RNA helixes and that both proteins make specific contacts with phosphates which are displaced from the sites of base-pair contact.

Base Composition↗
PubMed1994

Left atrial dilatation in constrictive pericarditis: a pre and post-operative echocardiographic study.

Thirty-three surgically proven cases of constrictive pericarditis were studied pre-operatively by echocardiography for left atrial dilatation and 18 of them underwent post-operative study at a mean follow-up period of 229 +/- 105 days. The degree of left atrial dilatation in these patients was compared with patients of restrictive cardiomyopathy (n = 8) and sex matched controls of similar age (n = 33). Significant left atrial dilatation was present in patients of constrictive pericarditis compared to controls and it was of a similar or greater degree compared to patients of restrictive cardiomyopathy (the left atrium to aorta ratio was 1.7 +/- 0.31, 1.53 +/- 0.18 and 1.07 +/- 0.1, in constrictive pericarditis, restrictive cardiomyopathy and controls, respectively, P = NS for constrictive pericarditis vs. restrictive cardiomyopathy and P < 0.001 for constrictive pericarditis vs. controls). There was no correlation of degree of left atrial dilatation with clinical and hemodynamic variables. Postoperatively, there was regression of left atrial size in patients with normal hemodynamics (n = 12, pre- vs. post-operative left atrium to aorta ratio 1.65 +/- 0.23 vs. 1.32 +/- 0.14, P < or = 0.001) and persisting or increasing left atrial dilatation in patients with persisting hemodynamic abnormality (n = 6, left atrium to aorta ratio 1.66 +/- 0.23 vs. 1.82 +/- 0.15, P = NS). We conclude that significant left atrial dilatation is a consistent echocardiographic feature of constrictive pericarditis. Hemodynamic normalization following successful pericardiectomy is associated with regression of atrial size.

Adolescent↗
PubMed1994

Ovarian steroid deprivation results in a reversible learning impairment and compromised cholinergic function in female Sprague-Dawley rats.

We hypothesized that estradiol (E2) serves as a neurotrophomodulatory substance for basal forebrain cholinergic neurons thought to be involved in learning and memory. Learning/memory was assessed using the two-way active avoidance paradigm and the Morris water task. Female Sprague-Dawley rats were either ovariectomized (OVX) or OVX for 3 weeks, followed by s.c. implantation of a Silastic pellet containing 17-beta E2 (E2 pellet), resulting in a replacement of E2 to physiological levels. Ovary-intact (INTACT) animals served as our positive control. Active avoidance behavior and choline acetyltransferase (ChAT) activity in the frontal cortex and hippocampus were assessed at 5 and 28 weeks postovariectomy while performance on the Morris water task and high-affinity choline uptake (HACU) were measured only at the 5-week time point. At the 5-week time point, E2 replacement caused a significant elevation in the level of active avoidance performance relative to OVX animals. At the 28-week time point, OVX animals demonstrated a significantly lower number of avoidances relative to controls (61%) whereas E2-pellet animals not only demonstrated superior performance relative to OVX animals but also showed an accelerated rate of learning. Morris water task performance, on the other hand, was not significantly affected by estrogenic milieu despite a trend towards better performance in the E2-pellet group. Neurochemical analyses revealed that 5 weeks of ovariectomy was sufficient to reduce HACU in both the frontal cortex and hippocampus by 24 and 34%, respectively, while E2 replacement was successful in elevating HACU relative to OVX animals in both regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Conjugated linoleic acid suppresses mammary carcinogenesis and proliferative activity of the mammary gland in the rat.

Conjugated linoleic acid (CLA) is a collective term which refers to a mixture of positional and geometric isomers of linoleic acid. It is naturally occurring in meat and dairy products. We have previously reported (Ip, C., Chin, S. F., Scimeca, J. A., and Pariza, M. W. Cancer Res., 51: 6118-6124, 1991) that 1% CLA in the diet suppressed mammary carcinogenesis in rats given a high dose (10 mg) of 7,12-dimethylbenz(a)anthracene. In the present study, dietary CLA between 0.05 and 0.5% was found to produce a dose-dependent inhibition in mammary tumor yield when fed chronically to rats treated with a lower dose (5 mg) of 7,12-dimethylbenz(a)anthracene. Short-term CLA feeding for 5 weeks, from weaning to the time of carcinogen administration at 50 days of age, also offered significant protection against subsequent tumor occurrence. This period corresponds to maturation of the mammary gland to the adult stage in the rat. The inhibitory response to short-term CLA exposure was observed with the use of 2 different carcinogens: 7,12-dimethylbenz(a)anthracene and methylnitrosourea. The fact that CLA was protective in the methylnitrosourea model suggests that it may have a direct modulating effect on susceptibility of the target organ to neoplastic transformation. The proliferative activity of the mammary epithelium was assessed by the incorporation of bromodeoxyuridine. Immunohistochemical staining results showed that CLA reduced the labeling index of the lobuloalveolar compartment, but not that of the ductal compartment of the mammary tree. Since the lobuloalveolar structures are derived from the terminal end buds which are the sites of carcinogenic transformation, the above finding is consistent with the bioassay data of tumor inhibition. Thus, changes in gland development and morphogenesis may be a locus of action of CLA in modulating mammary carcinogenesis. CLA is a unique anticarcinogen because it is present in foods from animal sources. Furthermore, its efficacy in cancer protection is manifest at dietary concentrations which are close to the levels consumed by humans.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1994

A computerized system for sequential analysis of aggregation of erythrocytes under dynamic conditions.

Aggregation of erythrocytes at hematocrit 5% in plasma, placed in a glass chamber, is analyzed under dynamic conditions by transmission of He-Ne laser light. The transmitted intensity (TI) signal which contains mean intensity superimposed with fluctuations caused by the movement of erythrocytes and formed aggregates, is detected by photodiode-amplifier assembly mounted on a stepper motor. The alignment of this detector for the maximum TI is achieved by a dedicated software. The TI data after A/D conversion is stored in a PC-AT for further processing. The parameters, aggregate size index, aggregate sedimentation time index, total time taken for completion of the process, process onset time index, total number of fluctuations, and asymmetry index are calculated sequentially for the entire data. A comparison of these parameters in blood samples obtained from healthy and diabetic subjects show the significance of these parameters.

Computer Systems↗
PubMed1994

A multiple fission-like mode of asexual reproduction in Blastocystis hominis.

A non-axenic and an axenic isolate of Blastocystis hominis have been induced to form cysts in vitro using an encystation medium. The morphology of the parasite at different time points was observed by scanning electron microscopy. In day-2 cultures the cysts were spherical and had a non-uniform, coarse outer surface around the body. A deep, pore-like opening was seen in some of the parasites. Most of the cysts from day-4 and day-6 cultures ruptured, revealing small, uniformly sized spherical bodies occurring in grape-like clusters. Acridine orange staining confirmed that these bodies were the progeny of Blastocystis hominis. A multiple fission-like reproduction process giving rise to many daughter Blastocystis occurs within the cyst.

Acridine Orange↗
PubMed1994

Clinico-pathological studies on experimental cryptococcal mastitis in goats.

Unilateral intramammary inoculation of 10 goats with Cryptococcus neoformans (2 x 10(6) yeast cells) resulted in the development of mastitis, with gross and microscopic lesions being restricted to the infected udder halves only and there was no dissemination of infection to the opposite uninfected udder halves as well as to other organs of the body. The experiment was continued for 40 days, with 2 animals each from the infected and control groups being killed on 5th, 10th, 20th, 30th and 40th day post-inoculation (DPI). Initial enlargement of the infected udder halves was followed by marked decrease in size leading to very small, firm and nodular udder halves. After infection, there was also sharp fall in the milk yield. Cryptococcal organisms were demonstrated in the mastitic milk and udder impression smears with special stains. C. neoformans was reisolated from the milk of the only infected udder halves up to 25th DPI. Microscopically, there was initially acute diffuse purulent mastitis which later on became chronic, characterised by marked infiltration of lymphocytes, macrophages, extensive fibrosis and development of multiple granulomas. The cryptococcal organisms could be demonstrated in the udder sections only up to 30th DPI. It is concluded that intramammary inoculation of Cryptococcus neoformans in goats leads to severe mastitis with sharp fall in milk yield.

Animals↗
PubMed1994

Ultrastructural changes during in vitro encystment of Blastocystis hominis.

The morphological changes occurring in Blastocystis hominis at different time points following in vitro encystment were studied by electron microscopy. The following stages of the parasite were sequentially seen: (a) the amoebic form, which was irregular in shape, with a majority of the organelles being concentrated at the condensed cytoplasmic region; (b) the pre-cystic form, which was rounded and had an electron-dense material forming a homogeneous wall around the central body; and (c) the cystic form, which had a very prominent, thick osmiophilic electron-dense wall, within which there were many inclusions and possibly reproductive granules. The amoebic form appeared to be an intermediate stage between the vacuolar form and the pre-cystic form, as this stage allowed the parasite to ingest bacteria to enhance encystment. The pre-cystic stage had previously been shown in experimental infection to be infective. The role of the cystic stage in producing infection is currently being investigated.

Animals↗
PubMed1994

Differentiation of the various stages of Blastocystis hominis by acridine orange staining.

Acridine orange staining differentiates the cystic and the central body forms of Blastocystis hominis and offers a very convenient and easy method to observe the internal structure of the parasite. Acridine orange stains the nuclei and the central body of the rounded vacuolar forms of the parasite bright and dull green, respectively. The colour changes to yellow and then to flaming red-orange when the rounded central body forms of the parasite become cystic.

Acridine Orange↗
PubMed1994

A behavioral measure of hand preference as opposed to hand skill.

Twenty-five self-professed left-handers and 21 self-professed right-handers were given a variety of performance tests to assess handedness, along with a preference inventory and a dichotic listening test of language lateralization. The performance tests included the Annett pegboard task, that Tapley and Bryden dot-filling tasks, and two procedures, the long pegboard and long dots tasks, that were intended to assess the point in space at which a particular unimanual movement became sufficiently awkward for one to shift to the other hand. All four of these performance tests differentiated between left-handers and right-handers, although the differences between handedness groups were somewhat larger when handedness was defined in terms of the preference inventory rather than on the basis of self-report. When the difference between preferred and non-preferred hands was examined, the best predictor of hand preference was the long pegboard task. Such a finding is consistent with the view that the long pegboard provides a behavioral measure of hand preference, while the pegs and dots tasks are more closely linked to specific skills. In addition, the correlations between individual preference items and the dichotic right-ear advantage suggest that language lateralization is related to rather different handedness measures than those usually used to define handedness. This finding would suggest that handedness and language lateralization are determined by somewhat different mechanisms.

Dichotic Listening Tests↗