Splenic abscess amenable to conservative therapy.
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Abscess↗
Biomedical subjects
A Agarwal
Publications and source records attributed to A Agarwal.
A p53-dependent S-phase checkpoint helps to protect cells from DNA damage in response to starvation for pyrimidine nucleotides.
Normal mammalian cells arrest primarily in G1 in response to N-(phosphonacetyl)-L-aspartate (PALA), which starves them for pyrimidine nucleotides, and do not generate or tolerate amplification of the CAD gene, which confers resistance to PALA. Loss of p53, accompanied by loss of G1 arrest, permits CAD gene amplification and the consequent formation of PALA-resistant colonies. We have found rat and human cell lines that retain wild-type p53 but have lost the ability to arrest in G1 in response to PALA. However, these cells still fail to give PALA-resistant colonies and are protected from DNA damage through the operation of a second checkpoint that arrests them reversibly within S-phase. This S-phase arrest, unmasked in the absence of the G1 checkpoint, is dependent on p53 and independent of p21/waf1.
Animals↗
B cell responses to a peptide epitope. VII. Antigen-dependent modulation of the germinal center reaction.
Germinal center responses to two analogous peptides, PS1CT3 and G32CT3, that differ in sequence only at one position within the B cell epitopic region were examined. In comparison with peptide PS1CT3, peptide G32CT3 elicited a poor germinal center response. By demonstrating equal facility of immune complexes with IgM and IgG Ab isotypes to seed germinal centers, we excluded differences in isotype profiles of early primary anti-PS1CT3 and anti-G32CT3 Ig as the probable cause. Quantitative differences in germinal center responses to the two peptides were also not due to either qualitative/quantitative differences in T cell priming or variation in the frequency of the early Ag-activated B cells induced. Rather, they resulted from qualitative differences in the nature of B cells primed. Analysis of early primary anti-PS1CT3 and anti-G32CT3 IgMs revealed that the latter population was of a distinctly lower affinity, implying the existence of an Ag affinity threshold that restricts germinal center recruitment of G32CT3-specific B cells. The impediment in anti-G32CT3 germinal center initiation could be overcome by making available an excess of Ag-activated Th cells at the time of immunization. This resulted in the appearance of a higher affinity population of G32CT3-specific B cells that, presumably, are now capable of seeding germinal centers. These data suggest that the strength of a germinal center reaction generated is Ag dependent. At least one regulatory parameter represents the quality of B cells that are initially primed.
Amino Acid Sequence↗
p21Waf1 inhibits the activity of cyclin dependent kinase 2 by preventing its activating phosphorylation.
Prostaglandin A2 (PGA2), a potent inhibitor of the growth of many cell types, inhibits G1 phase cyclin dependent kinases (cdk). Although PGA2 suppresses cyclin D1 and elevates p21Waf1 levels, it was the failure of cdk2 to become activated by phosphorylation which correlated best with growth inhibition. In kinetic studies, cdk2 activation was inhibited efficiently only if p21Waf1 levels increased prior to the activating phosphorylation; suggesting that p21Waf1 had blocked this phosphorylation. This model was confirmed in cells from p21Waf1 knockout mice where PGA2 was completely unable to block the activating phosphorylation of cdk2, or inhibit cdk2 activity. As expected, growth inhibition of p21Waf1(-/-) cells was not observed at PGA2 concentrations which inhibited cdk2 activity and growth of p21Waf1(+/+) cells.
3T3 Cells↗
Structural and functional heterogeneity among the zinc fingers of human MRE-binding transcription factor-1.
MRE-binding transcription factor-1 (MTF-1) activates the expression of metallothionein (MT) genes in mouse and human cells upon binding to one or more tandem metal-response elements (MREs; 5'-ctnTGCRCnCgGCCc) in the MT promoter. MTF-1 contains six Cys2-His2 zinc finger sequences. Previous work suggests that the zinc finger domain itself may function as a zinc sensor in zinc-activated expression of MTs. To obtain molecular insight into MTF-1 function, a recombinant fragment of MTF-1 containing only the zinc finger domain (denoted MTF-zf) has been purified using nondenaturing conditions and characterized with respect to zinc-binding properties, secondary structure, and DNA-binding activity. Different preparations of MTF-zf, following an anaerobic dialysis to quantify Zn(II) and reduced cysteine (by DTNB reactivity) content, reveal Zn(II)/MTF-zf stoichiometries ranging from 3.3 to 5.5 g at Zn(II) and 11-13 reduced thiolates (12 expected). Far-UV CD spectra reveal indistinguishable secondary structural content in all preparations, i.e., enough to fold just three of six zinc fingers of MTF-zf. Removal of additional zinc from MTF-zf gives rise to an insoluble apoprotein. Complex formation between a Zn5.5 MTF-zf and a coumarin-labeled MREd-containing oligonucleotide as monitored by changes in the anisotropy of the coumarin fluorescence gives a Kapp = 3.8 (+/-0.5) x 10(8) M-1 (pH 7.0, 0.20 M NaCl, 25 degreesC). Investigation of the salt type and concentration dependence of Kapp suggests significant contributions from both cation and anion release upon complex formation. Zn5.5 MTF-zf exhibits a large negative heat capacity of complex formation with MREd and can discriminate among DNA duplexes which have mutations deposited on either the TGCRC core or the C-rich side of the MREd. Air oxidation of Zn5.5 MTF-zf results in the reversible conversion of 6 of the 12 Cys thiolates to 3 disulfide bonds; as expected, this has no effect on the secondary structure of MTF-zf, but results in approximately 30-fold reduction in Kapp to approximately 1.2 x 10(7) M-1. In contrast, fully reduced Zn3.5 MTF-zf binds to the MREd with an affinity and [NaCl] dependence largely indistinguishable from those of Zn5.5 MTF-zf. The zinc fingers in MTF-zf are physically and functionally inequivalent. A subset (approximately 3-4) of zinc fingers plays a structural role in folding and high-affinity MREd binding, while one or more additional fingers have properties potentially consistent with a metalloregulatory role.
Amino Acid Sequence↗
Why cancer patients request disposal of cryopreserved semen specimens posttherapy: a retrospective study.
OBJECTIVE: To determine why patients with cancer stop storing semen in a sperm bank program. DESIGN: Retrospective study. SETTING: Hospital andrology laboratory. PATIENT(S): Cancer patients (n = 56) who discontinued sperm storage. INTERVENTION(S): A database of 342 patients with cryopreserved sperm was searched for disease diagnosis, marital status before and after diagnosis, type of therapy, number of specimens banked, interval between diagnosis and sperm banking, and postthaw semen characteristics. Patients discontinuing storage were surveyed. MAIN OUTCOME MEASUREMENT(S): Reasons for discontinuing storage and clinical correlation of the decision. RESULT(S): Reasons included patient death (n = 21); fertility but no plans for more children (n = 23); good sperm quality (n = 8); and no plans to have children (n = 4). Patients were similar in age, number of specimens, and interval between diagnosis and treatment, but they showed significant differences in type of treatment and time in the program. Cost of cryopreservation and specimen storage was not cited. CONCLUSION(S): Most patients decided to discontinue sperm banking because either they regained fertility or had improved semen quality. Sperm banking should be strongly recommended for all patients with malignant diseases who may wish to have children, even if they eventually decide that the specimens are not needed.
Adult↗
Accuracy of computer-assisted semen analysis in prefreeze and post-thaw specimens with high and low sperm counts and motility.
OBJECTIVES: We evaluated the accuracy and precision of computer-assisted semen analysis (CASAs) to determine whether variations in the CASA results were related to the type of analyzer or to the type of specimens analyzed. METHODS: Semen specimens were analyzed manually and by CASA before freezing and after thawing. Multiple ejaculates (220 semen specimens) from normal healthy donors (n = 8) and 131 semen specimens from 57 patients with cancer were assessed. RESULTS: The differences between CASA and manual sperm counts and percent motility were higher in prefreeze and post-thaw specimens at sperm concentrations of less than 20 x 10(6)/mL. The differences between CASA and manual results were significant for cancer patients, compared with normal donors (P <0.01). CONCLUSIONS: CASA results are unreliable at sperm counts of less than 20 x 10(6)/mL and post-thaw motility is generally underestimated by CASAs.
Cryopreservation↗
Identification of spermatozoa and round spermatids in the ejaculates of men with spermatogenic failure.
OBJECTIVES: As many as 10% of infertile men have azoospermia caused by spermatogenic failure or ductal obstruction. The histologic diagnoses associated with spermatogenic failure--Sertoli cell-only syndrome, maturation arrest, and hypospermatogenesis--do not necessarily represent global changes in the affected testis, as occasional seminiferous tubules may still produce mature germ cells. Intracytoplasmic sperm injection (ICSI) allows individual sperm that have been isolated from testicular tissue to fertilize oocytes. This study assessed whether mature germ cells (either round spermatids or spermatozoa) were present in the ejaculates of patients with spermatogenic failure. METHODS: All semen analyses performed at our tertiary care institution from January 1993 through December 1995 were reviewed to identify azoospermic men with spermatogenic failure. During this period, our laboratory employed Nuclear-Fast Red and picroindigocarmine staining (NF-PICS) of cytospin slides to identify rare spermatozoa and spermatids in otherwise azoospermic ejaculates. RESULTS: Of 3005 analyses reviewed, 20 azoospermic men whose infertility was solely attributable to spermatogenic failure were identified. The histologic diagnoses were germinal cell aplasia (n = 7), complete maturation arrest (n = 6), incomplete maturation arrest (n = 3), and hypospermatogenesis (n = 4). Using the NF-PICS technique, mature germ cells were identified in the ejaculates of 15 men (75%), and 9 men (45%) had fully formed spermatozoa present. CONCLUSIONS: In the clinical management of azoospermic infertile men, the NF-PICS technique may be used to identify men who have some degree of testicular spermatogenesis. This might obviate the need for the purely diagnostic testis biopsy that is performed before therapeutic biopsy for testicular sperm extraction in conjunction with ICSI.
Biopsy↗
Role of electron microscopy of sperm in the evaluation of male infertility during the era of assisted reproduction.
OBJECTIVES: To determine whether there are indications for and benefits from electron microscopy (EM) of sperm during the era of assisted reproductive technology. METHODS: The medical history, semen analyses, and EM findings of 55 patients with pure male-factor infertility were reviewed to determine: (1) which semen characteristics (seminal volume, sperm concentration, percent motility, and percent normal morphology) were associated with normal or abnormal ultrastructure as determined by EM, (2) whether EM findings correlated with success or failure of in vitro fertilization (IVF), and (3) whether EM could screen for potentially inheritable genetic disorders. RESULTS: Principal EM diagnoses were normal sperm ultrastructure (11 of 55; 20%), tail abnormalities (21 of 55; 38%), necrospermia (12 of 55; 22%), acrosomal defects (9 of 55; 16%), neck abnormalities (1 of 55; 2%), and incomplete maturation (1 of 55; 2%). Every patient with an abnormal EM study had impaired motility (33% or below) and abnormal morphology (13% or lower normal forms by World Health Organization criteria). The percentage of normal sperm morphology differed significantly across EM diagnoses (P < 0.0001). Differences in motility across the groups could not be detected because EM was only performed on patients with impaired motility. Although the partner of 1 patient with a normal EM study who underwent IVF achieved pregnancy, 11 with abnormal EM studies failed to establish a pregnancy by IVF. Finally, 5 (11%) of 44 patients with abnormal EM findings were diagnosed with primary ciliary dyskinesia directly as a result of EM. In addition, 3 (6.8%) of the 44 patients with abnormal EM findings were diagnosed with complete acrosomal loss. CONCLUSIONS: Patients with severely abnormal motility and morphology on routine semen analysis may benefit from EM study of sperm. Our data indicate that EM findings correlate with success or failure of IVF, and that EM can screen for potentially inheritable genetic disorders.
Humans↗
Treatment of erectile dysfunction after radical prostatectomy with sildenafil citrate (Viagra).
OBJECTIVES: To determine whether the response to the new oral medication, sildenafil citrate (Viagra), was influenced by the presence or absence of the neurovascular bundles, as recent reports on its success did not specify the efficacy of the drug in patients with erectile dysfunction after radical prostatectomy. METHODS: Baseline and follow-up data from 28 healthy patients presenting with erectile dysfunction after radical prostatectomy were obtained. Patients receiving any neoadjuvant/adjuvant hormones or adjuvant radiation therapy were excluded. Patients reported what their erectile status was before surgery, before sildenafil therapy, and after using a minimum of four doses of sildenafil. Both the patients and their spouses were interviewed using the Cleveland Clinic post-prostatectomy questionnaire, which includes questions about response to therapy, duration of intercourse, spousal satisfaction, side effects, and related topics. The patients were compared on the basis of the type of surgical procedure they had undergone-nerve sparing or non-nerve sparing. A positive response to sildenafil was defined as erection sufficient for vaginal penetration. RESULTS: Of the 15 patients who had bilateral nerve-sparing procedures, 12 (80%) had a positive response to sildenafil, with a mean duration of 6.92 minutes of vaginal intercourse. These 12 patients also reported a spousal satisfaction rate of 80%. All 12 of the responders had a positive response within the first three doses, and 10 of the 12 responded with the first or second dose. None of the 3 patients who had undergone a unilateral nerve-sparing procedure responded, nor did any of the 10 patients who had undergone a non-nerve-sparing procedure. The two most common side effects of the drug were transient headaches (39%) and abnormal color vision (11%). No patients discontinued the medication because of side effects. CONCLUSIONS: Successful treatment of erectile dysfunction in a patient after prostatectomy with sildenafil citrate may depend on the presence of bilateral neurovascular bundles. No patient who had undergone a non-nerve-sparing procedure responded. Whether patients who undergo unilateral nerve-sparing procedures will respond to sildenafil is still unclear because of the small number of patients in our study. These findings should encourage urologists to continue to perform and perfect the nerve-sparing approach. The ability to restore potency with an oral medication after radical prostatectomy will impact our discussion with the patient on the surgical morbidity of radical prostatectomy.
Enzyme Inhibitors↗
Relationship between creatine kinase levels and clinical diagnosis of infertility.
PURPOSE: The creatine kinase level indicates sperm maturity and correlates with the spermatozoal fertilizing potential. The relationship between creatine kinase levels in subfertile men and their clinical diagnosis was examined. METHODS: Patients with unexplained infertility (n = 34), varicocele (n = 20), postvasectomy reversal (n = 7), or cancer (n = 22) were included in this prospective clinical study. The control group consisted of healthy normal donors (n = 15). RESULTS: The median and interquartile range values of creatine kinase for each group were as follows: normal donors, 0.061 U/10(8) sperm (0.056 to 0.076 U/10(8) sperm); idiopathic male factor, 0.119 U/10(8) sperm (0.061 to 0.190 U/10(8) sperm); varicocele, 0.392 U/10(8) sperm (0.209 to 1.494 U/10(8) sperm); postvasectomy reversal, 0.589 U/10(8) sperm (0.425 to 4.043 U/10(8) sperm); and cancer, 0.068 U/10(8) sperm (0.047 to 0.168 U/10(8) sperm). Sperm creatine kinase levels were significantly higher in patients with varicocele compared to normal donors (P = 0.0001), cancer patients (P = 0.0002), and men with idiopathic infertility (P = 0.0009). Sperm concentration and creatine kinase level were inversely correlated in patients (r = -0.7, P < 0.001) but not in normal donors. CONCLUSIONS: Semen quality is poorer in subfertile patients with clinical varicocele and postvasectomy reversal than in cancer patients and patients with idiopathic male infertility. That the creatine kinase levels in cancer patients were similar to those of normal donors suggests that the final phase of spermatogenesis may not be altered in men with cancer; thus semen from these patients should be banked to ensure fertility after cancer treatment.
Creatine Kinase↗
Histopathological changes following the use of biological and synthetic glue for dural grafts: an experimental study.
Dural defects and CSF leaks are common neurosurgical problems and the search for a suitable sealant continues. The effects of such sealants on meninges and brain have not been previously studied. To study the effect of two commonly used sealants, burrholes were made in 20 rats, with the creation of dural tears which were covered with human cadaveric dura smeared with synthetic (cyanoacrylate) glue and gelatine (GRG) glue. Plain dura, placed in one of the burrholes served as a control. An intense inflammatory reaction was observed with both glues, which was found to persist till the eleventh week.
Adult↗
Acute suppurative thyroiditis with demonstrable distant primary focus: a report of two cases.
Two cases of acute suppurative thyroiditis occurring in a normal thyroid gland secondary to a demonstrable primary focus at a distant site are described and the role of interventional radiology in the management is discussed.
Acute Disease↗
Cryopreservation of human spermatozoa with pentoxifylline improves the post-thaw agonist-induced acrosome reaction rate.
Cryopreservation causes extensive damage to spermatozoa, thereby impairing their fertilizing ability. The purpose of this study was to determine if the direct addition of pentoxifylline to the seminal plasma before cryopreservation improved sperm motility and acrosome reaction. Semen specimens from 15 healthy volunteers were divided into two aliquots. One aliquot was treated by adding 5 mM pentoxifylline directly to the seminal plasma (treatment group) and the other aliquot received no treatment (control group). Both aliquots were then cryopreserved by using the liquid nitrogen freezing method. The percentage of motile spermatozoa and various motion characteristics were then evaluated by performing computer-assisted semen analysis. The sperm viability was determined with a supra-vital dye, Hoechst-33258, and the acrosome reaction (spontaneous and calcium ionophore-induced) was monitored using fluorescein isothiocyanate-conjugated peanut lectin (FITC-PNA) binding assays. Pentoxifylline treatment significantly increased the sperm motility, the amplitude of lateral head displacement, the hyperactivation status, and the frequency of spontaneous acrosome reactions before freezing (P < 0.05). After post-thaw, no difference in motion characteristics (except percentage motility) between treated and control groups were observed. Acrosome loss due to the freeze-thaw process was less in the pentoxifylline-treated group (P = 0.0003). In addition, the percentage of cryopreserved acrosome-intact spermatozoa that underwent further acrosome reactions in response to calcium-ionophore challenge was significantly higher in the treated group (P = 0.03). Pentoxifylline treatment before freezing improved the acrosome reaction to ionophore challenge in cryopreserved spermatozoa. Treatment with pentoxifylline appears to minimize sperm damage during the freeze-thaw process and may improve fertilization rates with assisted reproductive procedures such as intrauterine insemination or in-vitro fertilization.
Acrosome Reaction↗
Linoleyl hydroperoxide transcriptionally upregulates heme oxygenase-1 gene expression in human renal epithelial and aortic endothelial cells.
Atherogenic lipoproteins such as oxidized LDL are implicated in the pathogenesis of atherosclerosis and renal disease. Fatty acid hydroperoxides and phospholipids such as linoleyl hydroperoxide (LAox or 13-HPODE) and lysophosphatidylcholine (lyso-PC), abundant components of oxidized LDL, mediate the effects of atherogenic lipids. Oxidized LDL has been shown to induce heme oxygenase-1 (HO-1), a microsomal enzyme that is involved in heme detoxification and is a major endogenous source of carbon monoxide. HO-1 is also induced by many other stimuli that shift cellular redox. To identify the constituents and molecular mechanisms of oxidized LDL-mediated HO-1 induction, human renal epithelial cells and aortic endothelial cells were exposed to LAox and lyso-PC. Exposure to LAox (25 microM) showed an approximately 16-fold induction of HO-1 mRNA, whereas exposure to lyso-PC (25 microM) showed only an approximate 2.6-fold increase. Treatment with actinomycin-D (4 microM), a transcriptional inhibitor, as well as nuclear run-on assays, demonstrated that LAox-mediated HO-1 gene induction is dependent on de novo transcription. Cycloheximide did not affect LAox-mediated HO-1 mRNA induction, suggesting that new protein synthesis is not required for transcriptional induction. Transfection of a human HO-1 promoter-reporter gene construct showed that LAox upregulation of HO-1 occurs via mechanisms different from those of known inducers, heme and cadmium. These studies are the first demonstration that LAox induces HO-1 by transcriptional mechanisms and may have implications in the pathogenesis of cell injury in atherosclerosis and progressive renal disease.
Aorta↗
Cryptosporidium and HIV.
To study the importance of cryptosporidium as a diarrhoea causing pathogen (using stool samples), a cohort study was conducted from 14-9-1994 to 14-3-1995 on injecting drug users in Imphal. Stool samples were also collected from close contacts of the patients of human immunodeficiency virus (HIV) infected cryptosporidium diarrhoea cases and patients of paediatric medicine ward of Regional Institute of Medical Sciences (RIMS), Imphal. A total of 9 cases of cryptosporidium diagnosed out of 11 HIV infected diarrhoea cases and 2 out of 11 personal contacts were found to be positive. Two cases with history of close contacts with HIV infected cryptosporidium diarrhoea cases were not suffering from diarrhoea. One out of 7 paediatric diarrhoea cases was found to be positive. Five out of 9 HIV infected cryptosporidial cases died within one month of diagnosis of cryptosporidium. The remaining 4 had improved and diarrhoea was controlled. One of the 4 did not show cryptosporidium any more in the stool sample.
Adult↗
Pregnancy wastage in rural Varanasi: relationship with maternal nutrition and sociodemographic characteristics.
OBJECTIVE: To study the relationship between pregnancy wastage and matrnal undernutrition and other sociodemographic factors in rural Indian women. SETTING: Rural community of Varanasi. DESIGN: Longitudinal observation. METHOD: In 49 villages during 1988-92, 8111 pregnancies were registered to observe for wastage. The pregnancy outcome was correlated with various factors. RESULTS: There were 1321 abortions and 141 still births. Women's income < Rs. 250 per month, education < 10th class and protein intake < 50 g/day had significantly higher relative risks (RR) (4.1, 2.9 and 2.8, respectively) for abortions. Poor maternal nutrition was in additional important risk factor, for still births (RR 5.1 and 4.2 for maternal weight and height, respectively). A pregnancy interval over 2 years reduced both. CONCLUSION: Low socioeconomic status, chronic undernutrition and illiteracy in rural India are associated with high pregnancy wastage.
Abortion, Spontaneous↗
Nutritional status of rural non-pregnant non-lactating women in reproductive age.
OBJECTIVE: To find out the degree of current undernutrition in rural reproductive age women. SETTING: 49 villages of two adjoining rural blocks of Varanasi. METHODS: 6130 non-pregnant and non-lactating rural women in the age group 18-45 year were studied for sociodemographic characteristics and anthropometry, i.e., weight, height and midarm circumferance. Their percentiles for age and for weight for height were calculated by using cubic spline method. RESULTS: The women in 10th centile weighed < 38 kg and those in > 90th centile weighed 47-48 kg; 74.2% had weight < 45 kg. The 50th centile height ranged between 148-150 cm; 13.5% were < 145 cm. For mid arm circumference 50th and 90th centile values were around 22 and 24 cm, respectively. Mid arm circumference and height had significant linear correlation with weight. CONCLUSION: Around 50% rural UP women in pre-pregnancy state are undernourished. With age these rural women did not change in weight or mid-arm circumference.
Adolescent↗