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Biomedical subjects

B Lal

Publications and source records attributed to B Lal.

At least 19 recordsLinked to original sources

Comparative analysis of phenotypic and genotypic characteristics of two desulfurizing bacterial strains, Mycobacterium phlei SM120-1 and Mycobacterium phlei GTIS10.

AIM: To compare few phenotypic and genotypic characteristics of two desulfurizing bacterial strains, Mycobacterium phlei SM120-1 and Mycobacterium phlei GTIS10. METHODS AND RESULTS: In the present study, dibenzothiophene (DBT) desulfurizing activity, composition of fatty acids of cell membranes, DBT sulfone monoxygenase gene (bdsA) and the selection pressure applied during the growth and enrichment of the bacterial strains M. phlei SM120-1 and M. phlei GTIS10 were compared in our laboratory. The DBT desulfurization activity of M. phlei SM120-1 was found to be 0.17 +/- 0.02 micromol 2-HBP min(-1) (gram dry cell weight)(-1) and that of the bacterial strain M. phlei GTIS10 was 1.09 +/- 0.05 micromol 2-HBP min(-1) (gram dry cell weight)(-1). Fatty acid methyl ester analysis of cell membranes of these two bacterial strains in the presence of light gas oil showed that both the strains had different fatty acid profiles in their cell membranes. Comparison of the full gene sequences of the desulfurization gene bdsA in the two bacterial strains showed significant difference in the bdsA gene sequences. There was a significant difference observed in the selection pressure applied during the growth and enrichment of the two bacterial strains. CONCLUSIONS: The results of the comparative study of the bacterial strains, M. phlei SM120-1 and M. phlei GTIS10 showed that there were considerable differences in the phenotypic and genotypic characteristics of these two strains. SIGNIFICANCE AND IMPACT OF STUDY: The present study would broaden the understanding of biodesulfurization trait at intra-species level.

Amino Acid Sequence↗

In situ bioremediation potential of an oily sludge-degrading bacterial consortium.

A field-scale study was conducted in a 4000 m2 plot of land contaminated with an oily sludge by use of a carrier-based hydrocarbon-degrading bacterial consortium for bioremediation. The land belonged to an oil refinery. Prior to this study, a feasibility study was conducted to assess the capacity of the bacterial consortium to degrade oily sludge. The site selected for bioremediation contained approximately 300 tons of oily sludge. The plot was divided into four blocks, based on the extent of contamination. Blocks A, B, and C were treated with the bacterial consortium, whereas Block D was maintained as an untreated control. In Block A, at time zero, i.e., at the beginning of the experiment, the soil contained as much as 99.2 g/kg of total petroleum hydrocarbon (TPH). The application of a bacterial consortium (1 kg carrier-based bacterial consortium/10 m2 area) and nutrients degraded 90.2% of the TPH in 120 days, whereas in block D only 16.8% of the TPH was degraded. This study validates the large-scale use of a carrier-based bacterial consortium and nutrients for the treatment of land contaminated with oily sludge, a hazardous hydrocarbon waste generated by petroleum industry.

Acinetobacter↗

Evaluation of inoculum addition to stimulate in situ bioremediation of oily-sludge-contaminated soil.

A full-scale study evaluating an inoculum addition to stimulate in situ bioremediation of oily-sludge-contaminated soil was conducted at an oil refinery where the indigenous population of hydrocarbon-degrading bacteria in the soil was very low (10(3) to 10(4) CFU/g of soil). A feasibility study was conducted prior to the full-scale bioremediation study. In this feasibility study, out of six treatments, the application of a bacterial consortium and nutrients resulted in maximum biodegradation of total petroleum hydrocarbon (TPH) in 120 days. Therefore, this treatment was selected for the full-scale study. In the full-scale study, plots A and B were treated with a bacterial consortium and nutrients, which resulted in 92.0 and 89.7% removal of TPH, respectively, in 1 year, compared to 14.0% removal of TPH in the control plot C. In plot A, the alkane fraction of TPH was reduced by 94.2%, the aromatic fraction of TPH was reduced by 91.9%, and NSO (nitrogen-, sulfur-, and oxygen-containing compound) and asphaltene fractions of TPH were reduced by 85.2% in 1 year. Similarly, in plot B the degradation of alkane, aromatic, and NSO plus asphaltene fractions of TPH was 95.1, 94.8, and 63.5%, respectively, in 345 days. However, in plot C, removal of alkane (17.3%), aromatic (12.9%), and NSO plus asphaltene (5.8%) fractions was much less. The population of introduced Acinetobacter baumannii strains in plots A and B was stable even after 1 year. Physical and chemical properties of the soil at the bioremediation site improved significantly in 1 year.

Acinetobacter↗

Effect of Trigonella foenum-graecum (fenugreek) seeds on glycaemic control and insulin resistance in type 2 diabetes mellitus: a double blind placebo controlled study.

OBJECTIVES: To evaluate the effects of Trigonella foenum-graecum (fenugreek) seeds on glycemic control and insulin resistance, determined by HOMA model, in mild to moderate type 2 diabetes mellitus we performed a double blind placebo controlled study. METHODS: Twenty five newly diagnosed patients with type 2 diabetes (fasting glucose < 200 mg/dl) were randomly divided into two groups. Group I (n=12) received 1 gm/day hydroalcoholic extract of fenugreek seeds and Group II (n=13) received usual care (dietary control, exercise) and placebo capsules for two months. RESULTS: At baseline both the groups were similar in anthropometric and clinical variables. Oral glucose tolerance test, lipid levels, fasting C-peptide, glycosylated haemoglobin, and HOMA-model insulin resistance were also similar at baseline. In group 1 as compared to group 2 at the end of two months, fasting blood glucose (148.3 +/- 44.1 to 119.9 +/- 25 vs. 137.5 +/- 41.1 to 113.0 +/- 36.0) and two hour postglucose blood glucose (210.6 +/- 79.0 to 181.1 +/- 69 vs. 219.9 +/- 41.0 to 241.6 +/- 43) were not different. But area under curve (AUC) of blood glucose (2375 +/- 574 vs 27597 +/- 274) as well as insulin (2492 +/- 2536 vs. 5631 +/- 2428) was significantly lower (p < 0.001). HOMA model derived insulin resistance showed a decrease in percent beta-cell secretion in group 1 as compared to group 2 (86.3 +/- 32 vs. 70.1 +/- 52) and increase in percent insulin sensitivity (112.9 +/- 67 vs 92.2 +/- 57) (p < 0.05). Serum triglycerides decreased and HDL cholesterol increased significantly in group 1 as compared to group 2 (p < 0.05). CONCLUSIONS: Adjunct use of fenugreek seeds improves glycemic control and decreases insulin resistance in mild type-2 diabetic patients. There is also a favourable effect on hypertriglyceridemia.

Adult↗

Prevalence of coronary risk factors among Indian physicians.

BACKGROUND AND OBJECTIVES: Prevalence of coronary risk factors has not been well studied in Indian physicians, therefore, to determine prevalence of selected lifestyle and biochemical coronary risk factors we performed this study. METHODS: Physicians attending a national conference were invited to participate. Of the 1000 questionnaires circulated 256 physicians (221 males, 35 females) responded and were examined for presence of smoking, obesity, truncal obesity, hypertension and ECG abnormalities. Two hundred and thirty four physicians (91.4%, 203 males, 31 females) underwent a fasting blood examination for determination of glucose and lipid profile. Subjects were divided into three age groups: Group I aged < 40 years; Group II 40-49 years; and Group III > or = 50 years. RESULTS: Mean age of the study population was 41.7 +/- 9.3 years (range 26-70). There was no significant difference in the distribution of height, weight, body-mass index (BMI), waist and hip circumference and waist-hip ratio (WHR) in different age-groups. Systolic blood pressure (BP) and fasting glucose and triglyceride levels increased with age in both males and females, while diastolic BP, total - LDL and HDL cholesterol levels did not change. Smoking or tobacco use was seen in five males (2.3%). Prevalence of obesity (BMI > or = 25.0 kg/m2) was in 104 (48.6%) males and 18 (51.4%) females. Truncal obesity diagnosed by WHR > 0.9 in males and > 0.8 in females was in 160 (72.4%) males and 23 (65.7%) females and a large waist circumference, > or = 100 cm in males and > or = 90 cm in females, was in 58 (26.2%) males and 7 (20.0%) females. Hypertension (> or = 140/90) was in 74 (33.5%) males and 7 (20%) females. A high prevalence of diabetes diagnosed using fasting blood glucose > or = 126 mg/dl or previous history was noted in males 19 (9.4%), females 4 (12.9%). Prevalence of high total cholesterol levels > or = 200 mg/dl was in 91 (44.8%) males and 10 (32.3%) females. High LDL cholesterol level (> or = 100 mg/dl) was in 144 (70.9%) males and 22 (70.9%) females and LDL levels > or = 130 mg/dl in 70 (34.5%) males and 9 (29.0%) female physicians. High triglyceride levels (> or = 200 mg/dl) were in 38 (18.7%) males and 4 (12.9%) females. Electrocardiographic abnormalities were present in seven subjects: Q-waves in two males and one female and ST-T changes in three males and one female. BMI correlated significantly (p < 0.05) with systolic BP, fasting glucose, and triglycerides and WHR with systolic BP, diastolic BP, fasting glucose, cholesterol and triglyceride levels. CONCLUSIONS: There is a high prevalence of obesity, truncal obesity, hypertension and hypercholesterolaemia in Indian physicians while smoking, low HDL cholesterol and hypertriglyceridemia is low. The overall coronary risk is lower among Indian physicians as compared to previous Indian population studies.

Adult↗

Glioma inhibition by HGF/NK2, an antagonist of scatter factor/hepatocyte growth factor.

Strategies that antagonize growth factor signaling are attractive candidates for the biological therapy of brain tumors. HGF/NK2 is a secreted truncated splicing variant and potential antagonist of scatter factor/hepatocyte growth factor (SF/HGF), a multifunctional cytokine involved in the malignant progression of solid tumors including glioblastoma. U87 human malignant glioma cells that express an autocrine SF/HGF stimulatory loop were transfected with the human HGF/NK2 cDNA and clonal cell lines that secrete high levels of HGF/NK2 protein (U87-NK2) were isolated. The effects of HGF/NK2 gene transfer on the U87 malignant phenotype were examined. HGF/NK2 gene transfer had no effect on 2-dimensional anchorage-dependent cell growth. In contrast, U87-NK2 cell lines were approximately 20-fold less clonogenic in soft agar and approximately 4-fold less migratory than control-transfected cell lines. Intracranial tumor xenografts derived from U87-NK2 cells grew much slower than controls. U87-NK2 tumors were approximately 50-fold smaller than controls at 21 days post-implantation and HGF/NK2 gene transfer resulted in a trend toward diminished tumorigenicity. This report shows that the predominant effect of transgenic HGF/NK2 overexpression by glioma cells that are autocrine for SF/HGF stimulation is to inhibit their malignant phenotype.

Agar↗

Role of curcumin in idiopathic inflammatory orbital pseudotumours.

The present report, describes for the first time the clinical efficacy of curcumin, the active constituent of rhizomes of Curcuma longa, in the treatment of patients suffering from idiopathic inflammatory orbital pseudotumours. Curcumin was administered orally at a dose of 375 mg/3 times/day orally for a period of 6-22 months in eight patients. They were followed up for a period of 2 years at 3 monthly intervals. Five patients completed the study, out of which four recovered completely and in one patient the swelling regressed completely but some limitation of movement persisted. No side effect was noted in any patient and there was no recurrence. It is suggested that curcumin could be used as a safe and effective drug in the treatment of idiopathic inflammatory orbital pseudotumours.

Administration, Oral↗

Reversion of human glioblastoma malignancy by U1 small nuclear RNA/ribozyme targeting of scatter factor/hepatocyte growth factor and c-met expression.

BACKGROUND: Expression of scatter factor (SF), also known as hepatocyte growth factor (HGF), and its receptor, c-met, is often associated with malignant progression of human tumors, including gliomas. Overexpression of SF/HGF in experimental gliomas enhances tumorigenicity and tumor-associated angiogenesis (i.e., growth of new blood vessels). However, the role of endogenous SF/HGF or c-met expression in the malignant progression of gliomas has not been examined directly. In this study, we tested the hypothesis that human glioblastomas can be SF/HGF-c-met dependent and that a reduction in endogenous SF/HGF or c-met expression can lead to inhibition of tumor growth and tumorigenicity. METHODS: Expression of the SF/HGF and c-met genes was inhibited by transfecting glioblastoma cells with chimeric transgenes consisting of U1 small nuclear RNA, a hammerhead ribozyme, and antisense sequences. The effects of reduced SF/HGF and c-met expression on 1) SF/HGF-dependent induction of immediate early genes (c-fos and c-jun), indicative of signal transduction; 2) anchorage-independent colony formation (clonogenicity), an in vitro correlate of solid tumor malignancy; and 3) intracranial tumor formation in immunodeficient mice were quantified. Statistical tests were two-sided. RESULTS: Introduction of the transgenes into glioblastoma cells reduced expression of the SF/HGF and c-met genes to as little as 2% of control cell levels. Reduction in c-met expression specifically inhibited SF/HGF-dependent signal transduction (P<.01). Inhibition of SF/HGF or c-met expression in glioblastoma cells possessing an SF/HGF-c-met autocrine loop reduced tumor cell clonogenicity (P =.005 for SF/HGF and P=.009 for c-met) and substantially inhibited tumorigenicity (P<.0001) and tumor growth in vivo (P<.0001). CONCLUSIONS: To our knowledge, this is the first successful inhibition of SF/HGF and c-met expression in a tumor model directly demonstrating a role for endogenous SF/HGF and c-met in human glioblastoma. Our results suggest that targeting the SF/HGF-c-met signaling pathway may be an important approach in controlling tumor progression.

Animals↗

Efficacy of curcumin in the management of chronic anterior uveitis.

Curcumin, obtained from rhizomes of Curcuma longa, was administered orally to patients suffering from chronic anterior uveitis (CAU) at a dose of 375 mg three times a day for 12 weeks. Of 53 patients enrolled, 32 completed the 12-week study. They were divided into two groups: one group of 18 patients received curcumin alone, whereas the other group of 14 patients, who had a strong PPD reaction, in addition received antitubercular treatment. The patients in both the groups started improving after 2 weeks of treatment. All the patients who received curcumin alone improved, whereas the group receiving antitubercular therapy along with curcumin had a response rate of 86%. Follow up of all the patients for the next 3 years indicated a recurrence rate of 55% in the first group and of 36% in the second group. Four of 18 (22%) patients in the first group and 3 of 14 patients (21%) in the second group lost their vision in the follow up period due to various complications in the eyes, e.g. vitritis, macular oedema, central venous block, cataract formation, glaucomatous optic nerve damage etc. None of the patients reported any side effect of the drug. The efficacy of curcumin and recurrences following treatment are comparable to corticosteroid therapy which is presently the only available standard treatment for this disease. The lack of side effects with curcumin is its greatest advantage compared with corticosteroids. A double blind multi-centric clinical trial with this drug in CAU is highly desirable to further validate the results of the present study.

Adult↗

Plasmodium falciparum: drug-resistant malaria complicating leukemias and lymphomas in children.

The present communication deals with drug-resistant Plasmodium falciparum malaria complicating hematologic malignancies (leukemias, n = 24, and lymphomas, n = 7) in children. Of 50 cases of hematologic malignancies, 31 patients were microscopically diagnosed as having P. falciparum infection (MP +). Initially, all the patients were treated with chloroquine. The results of primary treatment showed chloroquine resistance in 16 (51. 62%) cases. Of these 16 chloroquine-resistant cases, 13 were secondarily treated with a combination of pyrimethamine plus sulfamethopyrazine. The results of secondary treatment also revealed resistance to pyrimethamine plus sulfamethopyrazine in 6 of 13 (46. 10%) cases. The 6 pyrimethamine plus sulfamethopyrazine-resistant P. falciparum cases were finally cured by quinine therapy, against which no resistance was encountered. Conversely, in the control group comprising 38 cases of P. falciparum without malignancy, the incidence of chloroquine resistance was found in only 9 cases, which is rather low (23.70%). Of these 7 chloroquine-resistant cases, 5 were found to be sensitive to pyrimethamine plus sulfamethopyrazine treatment, while the 2 nonresponders were finally cured with quinine. The overall results of this study show a high prevalence of chloroquine resistance among clinical cases of falciparum malaria (25/69; 30.6%). Among the nonresponders (n = 20) 40% of cases were also resistant to the pyrimethamine plus sulfamethopyrazine combination. There was no resistance to quinine.

Adolescent↗

Hydroxyacyl derivatives of forskolin--their positive inotropic activity.

Using appropriate protection and deprotection sequence novel hydroxyacyl chains of the type CO(CH2)nOH are synthesized and are utilized to develop new analogues of forskolin. Several compounds showed good positive inotropic activity. Compound 12 is almost 10 times more active than forskolin (EC50 = 0.002 microgram/ml).].

Acylation↗

In search of novel water soluble forskolin analogues for positive inotropic activity.

Using the novel lead from hydroxy acetyl substituted forskolin analogues, such as 7 beta-hydroxyacetyl-7 beta-deacetyl forskolin or 6 beta-hydroxyacetyl forskolin, a number of water soluble omega-amino acyl derivatives were synthesized. Two such compounds 6 and 18 showed better in vitro activity but failed to show in vivo activity.

Acetylation↗

Detection of anti-PPD IgG antibody and PPD-induced delayed type hypersensitivity in anterior uveitis patients.

Present study relates to the results of anti-PPD IgG, anti-A60 and antinuclear antibodies and PPD-induced delayed type hypersensitivity (DTH) in 17 anterior uveitis, (AU) patients. Results of anti-PPD IgG assay revealed detection of higher mean antibody level (O.D. 0.11 +/- 0.06) compared to healthy controls (O.D. 0.04 +/- 0.03), other eye disease controls (O.D. 0.05 +/- 0.003) and leprosy controls (O.D. 0.03 +/- 0.03). Anti-A60 IgM antibody assay revealed insignificant differences in mean antibody levels between various groups. Four of 17(23.5%) AU patients and 1(5.8%) subject each, belonging to other eye disease and healthy control groups had raised anti-nuclear antibody index. Findings of PPD skin test revealed detection of moderate to strong (2 to 4+) reactivity in 14 (82%). AU patients. Conversely, 13(76%) healthy controls and 8(47%) other eye disease controls gave mild (1+) reactivity. Results of this study suggested possible role of hypersensitivity to mycobacterial antigens in pathogenesis of anterior uveitis.

Antibodies, Antinuclear↗

Endothelial cell-based cytokine gene delivery inhibits 9L glioma growth in vivo.

Malignant brain neoplasms present great therapeutic challenges due to their extremely aggressive behavior and relative isolation by the blood-brain and blood-tumor barriers. Endothelial cells may be versatile platforms for delivering genes to solid tumors by virtue of their location at blood-tissue interfaces and their proliferation in response to endothelial mitogens produced by tumors. Immortalized rat brain endothelial cells that express the E. coli lacZ reporter gene and the gene for murine interleukin-2 (RBEZ-IL2) were co-inoculated with 9L glioma cells to Fisher rats to examine the effects of endothelial cell-based cytokine delivery on glioma growth in vivo. 9L glioma growth was not affected by the implantation of control RBEZ cells. The growth of subcutaneous and intracranial 9L gliomas was significantly inhibited by RBEZ-IL2 cells (P < 0.005 and P < 0.01, respectively) when compared to control transfected RBEZ cells. Rats receiving intracranial 9L glioma cells with RBEZ-IL2 cells showed increased survival (P < 0.001). Histologic and immunohistologic analysis showed enhanced activation of microglia/macrophages and CD8-positive T lymphocytes and/or natural killer cells within brain at sites of 9L inoculation with RBEZ-IL2 cells. This report establishes that immortalized endothelial cells can be used for cytokine gene delivery and to activate anti-tumor host responses to experimental gliomas within the central nervous system.

Animals↗

Role of anion exchange and thiol groups in the regulation of potassium efflux by lead in human erythrocytes.

Pb2+ is thought to enter erythrocytes through anion exchange (AE) and to remain in the cell by binding to thiol groups. To define the role of AE mechanisms and thiol groups in Pb2+ toxicity, we studied the effects of drugs and conditions that modify AE and that modify thiol groups on the ability of Pb2+ to stimulate potassium efflux as measured with 86Rb. The most potent stimulation of 86Rb efflux by Pb2+ occurred when conditions were optimal for the AE mechanism--that is, when bicarbonate was included in the buffer or a buffer made with Nal or NaCl rather than NaClO4 or NaNO3 was used. Furthermore, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid and 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfuonic acid, potent inhibitors of the AE mechanism, completely inhibited stimulation of the 86Rb efflux by Pb2+. These conditions or inhibitors did not affect stimulation of the 86Rb efflux by ionomycin plus Ca2+. A role for Ca2+ channels was dismissed because the inorganic Ca2+ channel blockers, Cd2+ or Mn2+, did not prevent stimulation of 86Rb efflux by Pb2+ but did inhibit stimulation by ionomycin plus Ca2+. 86Rb efflux was more sensitive to Pb2+ if erythrocytes were treated for 15 min with thiol-modifying reagents that enter cells, such as iodoacetamide, N-ethylmaleimide, or dithiothreitol, than to reduced glutathione, a thiol-modifying reagent that is not permeable to the cell. Thus, in erythrocytes the AE mechanism and internal thiol groups are critical factors that affect the stimulation of a Ca(2+)-dependent process by Pb2+.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Chest wall hamartoma in an adult: an unusual chest wall tumor.

An asymptomatic intrathoracic mass was demonstrated on imaging studies in an adult. Operative removal was accomplished and it proved to be an extrapulmonary hamartoma arising from the soft tissue of the chest wall. This represents a rare location of this benign tumor in an adult.

Adult↗

Degradation of crude oil by Acinetobacter calcoaceticus and Alcaligenes odorans.

Two types of Indian crude oil (Bombay High and Gujarat) were tested for their biodegradability by Acinetobacter calcoaceticus and Alcaligenes odorans. Acinetobacter calcoaceticus S30 and Alc. odorans P20 degraded Bombay High crude oil by 50% and 45%, while only 29% and 37% of Gujarat crude oil (heavy crude oil) was degraded by these isolates, respectively. Acinetobacter calcoaceticus and Alc. odorans in combination degraded 58% and 40% of Bombay High and Gujarat crude oils, respectively, which were significantly higher than that of by individual cultures. Acinetobacter calcoaceticus S30 degraded more of the alkanes fraction than the aromatics fraction of both crude oils. GC fingerprinting of alkane fraction showed major degradation of heptadecane (C17), octadecane (C18), nonadecane (C19), eicosane (C20), docosane (C22), tricosane (C23) and tetracosane (C24) of crude oil, while the Alc. odorans P20 degraded alkanes and aromatics equally. The asphaltenic component increased in both types of crude oil after biodegradation . The two strains grew very well on n-alkane up to C33 as well as on pristane (branched-chain alkane) but could not grow on cycloalkanes. Acinetobacter calcoaceticus S30 could not grow on pure polycyclic aromatic hydrocarbon (PAH) compounds except naphthalene but Alc. odorans P20 could grow on anthracene, phenanthrene, dibenzothiophene, fluorene, fluoranthene, pyrene and chrysene.

Acinetobacter calcoaceticus↗

Post-mastectomy seroma: a new look into the aetiology of an old problem.

Postmastectomy seroma (PMS) remains an unresolved quandary as the risk factors for its formation have still not been identified. Sixty-four consecutive female patients undergoing Halsted mastectomy were prospectively studied for this purpose. The risk factors that were studied included age and weight, diabetes, hypertension, tumour (size and location) and nodal (positive or negative) status and the use of perioperative blood transfusion. Additionally, the effect of PMS on total hospital drainage (THD), post-operative hospital stay and other wound-related complications were assessed. The incidence of PMS in this study was 28%, and those patients with PMS were designated as group A while the remaining patients were designated as group B. Group A patients were significantly older (P < 0.02) and heavier (P < 0.05) than group B patients; also, there were significantly (P < 0.0001) greater number of hypertensive patients in group A than in group B. In terms of other risk factors the two groups were statistically similar. Group A patients had significantly greater (P < 0.01) amount of THD than group B patients. Although no septic complications were observed as a result of seroma, its presence significantly (P < 0.0001) lengthened the post-operative hospital stay in group A patients. It is concluded that (1) PMS is a common problem, (2) its occurrence significantly increases the post-operative hospital stay, and (3) hypertension is perhaps the most important risk factor for its causation. Hypertension contributes to seroma formation, probably by way of prolonged oozing from the large mastectomy wound.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗