THE STUDY OF LIVER AND KIDNEY FUNCTION WITHIN COVID-19 PATIENTS.

Goal -the existing data on the coronavirus disease 2019 (COVID-19) revealed the impact of the disease on different organs such as the liver, the heart, the kidneys, etc. In our study, we evaluated the clinical characteristics of COVID-19 in patients with abnormal liver and kidney test results in recovered and dead patients. The number of the covid-19 patients was - 289. The gold standard method of polymerase chain reaction (PCR) was used to detect the infection of Covid-19. Blood plasma was used as research material. For The Determination of ALT, AST, and Creatinine, the Analyzer was the Prestige 24i - optimized, modified method according to the International Federation of Clinical Chemistry (IFCC), without pyridoxal phosphate; For the statistical analysis were used the Graphed Prism (Version 8.0). Studies have revealed an increase in AST and ALT compared with the reference value. A small difference was also found between those who died and those who recovered; A change in creatinine levels was also found. Men have a high level of Creatinine (~4.28 times) compared to the reference level, and in women, the level of Creatinine is ~2.18 times higher than the reference level. We have thought that according to our results, the high creatinine levels may have some risk impact for the severity of COVID-19 disease.

THE STUDY OF VDR FOKI RS2228570 SNP IN AUTOIMMUNE THYROIDITIS.

Autoimmune thyroid disease (AITD) is a common organ-specific autoimmune disease. A strong influence of genetic and epigenetic modifications has been demonstrated to take part in the development and progression of autoimmune thyroid diseases. The linkage between the Vitamin D receptor (VDR) polymorphism and several autoimmune disorders, including the AITD. In this article, we aim to investigate the Frequency of VDR Fokl (rs2228570) genotypes (CC, CT, TT) and alleles (C,T) in autoimmune thyroiditis. The investigation of VDR Fokl (rs2228570) was conducted on 150 samples (control (75 healthy women) and diseased women (75 diseased with autoimmune thyroiditis)) patients from the Adjara (Georgia) Population. It also examined some clinical and laboratory characteristics of the study population. Autoimmune thyroiditis's disease was diagnosed by measuring blood antibodies, determining the level of thyroperoxidase, and conducting an ultrasound examination. Anti-TPO and TSH were studied using the ELISA method. The genomic DNA was extracted from the peripheral blood. The polymerase chain reaction was evaluated to examine the VDR Fokl rs2228570 SNP polymorphism. According to VDR Fokl (rs2228570) genotypes (CC; CT, TT) frequency, in the control group, the Frequency of CC-genotype is 48%, CT-heterozygous genotype is 29.33%, and TT-genotype is 22.67%; in the diseased population, the Frequency of CC-genotype is 57.33%, CT-genotype is 34.67%, and TT-genotype is 8%. According to VDR Fokl (rs2228570) alleles (C, T), the Frequency of the C-allele is high, and the Frequency of the T-allele is low in both populations. The Frequency of the CC and CT genotypes of VDR Fokl (rs2228570) is high in the population with autoimmune thyroiditis compared to the control group; the TT genotype is relatively low in the population suffering from autoimmune thyroiditis; According to VDR Fokl (rs2228570) alleles (C, T), the Frequency of C-allele is high both population.

Evaluating the effects of high-oil rapeseed cake or natural additives on methane emissions and performance of dairy cows.

We evaluated the potential of feeding high-oil rapeseed cake or natural additives as rumen modifiers on enteric methane (CH4) emissions, nutrient utilization, performance, and milk fatty acid (FA) profile of dairy cows. Eight Nordic Red dairy cows averaging (mean ± SD) 81 ± 21 d in milk and 41.0 ± 1.9 kg of milk yield at the beginning of the study were randomly assigned to a replicated 4 × 4 Latin square design with 21-d periods. Treatments comprised grass silage-based diets (45:55 forage to concentrate ratio on dry matter basis) including (1) control containing 19.3% rapeseed meal (CON), (2) CON with full replacement of rapeseed meal with rapeseed cake (RSC), (3) supplementation of CON with 50 g/d of yeast hydrolysate product plus coniferous resin acid-based compound (YHR), and (4) supplementation of CON with 20 g/d of combination of garlic-citrus extract and essential oils in a pellet (GCE). Apparent total-tract digestibility was measured using total collection of feces, and CH4 emissions were measured in respiratory chambers on 4 consecutive days. Data collected during d 17 and 21 in each period were used for ANOVA analysis using a mixed model. Treatments did not affect dry matter intake (DMI), whereas feeding RSC increased crude protein and ether extract digestibility compared with the other diets. Emissions of CH4 per day, per kilogram of DMI, and per kilogram of energy-corrected milk, and gross energy intake were lower for RSC compared with other diets. We found no effect of YHR on daily CH4 emissions, whereas CH4 yield (g of CH4/kg of DMI or as percentage of gross energy intake) decreased with GCE compared with CON. Treatments did not influence energy balance. Further, RSC reduced the proportion of N intake excreted in feces, and YHR improved N balance compared with CON diet. Feeding RSC resulted in greatest yields of milk and energy-corrected milk, and feed efficiency. Relative to the CON diet, RSC decreased saturated FA by 10% in milk fat by increasing cis-monounsaturated FA but also increased the proportion of trans FA. Proportion of odd- and branched-chain FA increased with GCE and YHR compared with CON. We conclude that replacing rapeseed meal by rapeseed cake decreased CH4 emissions, whereas YHR or GCE had no effect on CH4 emissions in this study.

Modulation in the acidity constant of acridine dye with cucurbiturils: stimuli-responsive pKa tuning and dye relocation into live cells.

The noncovalent host-guest interactions of the cationic (AcH+) and neutral (Ac) forms of an acridine dye with macrocyclic hosts such as cucurbit[7]uril (CB7) and cucurbit[8]uril (CB8) have been investigated to evaluate the effect of cavity size on the photophysical properties and the protolytic equilibrium of the acridine dye. The cationic form undergoes significant complexation with CB7 (Keq = 106 M-1), causing a sharp decrease in the fluorescence intensity, whereas the neutral Ac form of the dye undergoes weak complexation with CB7 (Keq = 103 M-1) and the binding constant is lowered by three orders of magnitude compared to that of the CB7-AcH+ system. The Job plot revealed that both forms form a 1 : 1 complex with CB7. On the other hand, the AcH+ form shows strong emission quenching on interaction with CB8 and the formation of the 1 : 2 CB8 : AcH+ complex has been confirmed from the Job plot. The strong affinity of CB7 and CB8 to the protonated form resulted in a large upward pKa shift (ΔpKa ∼ 3.4 units for CB7 and ∼1.3 units for CB8) in the dye. Taking advantage of the above modulations in the fluorescence and pKa values, adamantylamine-induced fluorescence regeneration, controlled pKa tuning and dye relocation from the CB7 cavity to cell lines have been established for the first time, which find potential applications in fluorescence off-on sensing and drug delivery.

Metal ion-induced supramolecular pKa tuning and fluorescence regeneration of a p-sulfonatocalixarene encapsulated neutral red dye.

The host-guest interactions and the consequent modulation in the prototropic equilibrium of a phenazine dye, neutral red, with p-sulfonatocalix[4]arene (SCX4) and p-sulfonatocalix[6]arene (SCX6) macrocyclic hosts have been investigated. Both the neutral (NR) and cationic (NRH+) forms of dyes formed inclusion complexes with SCX6, with a larger binding constant for the latter (K = 8.6 × 105 M-1versus 4.8 × 103 M-1) due to the cation receptor behavior of the calixarenes. The distinct differences in the binding constant of NR and NRH+ provided a finite tuning of pKa between 6.5 and 8.8, through a competitive binding with metal ions. Importantly, the fluorescence quenching observed in the SCX-neutral red interactions stands in contrast to the fluorescence enhancement observed with other macrocyclic hosts, such as ß-cyclodextrin and cucurbit[7]uril. This is due to the unique portal stacking interaction of NRH+ with the SCXs, compared to the axial inclusion geometry documented for the other macrocycles. The electron transfer from the SCX to the neutral red dye is adjudged to be the effective excited-state relaxation pathway leading to fluorescence quenching. In combination with the metal-ion induced fluorescence regeneration and tuning the pKa value, the SCX-neutral red system finds potential applications in drug delivery, photodynamic therapy, catalysis, and sensor applications.

HIV status and viral loads among men testing positive for rectal gonorrhoea and chlamydia, Maricopa County, Arizona, USA, 2011-2013.

OBJECTIVES: Men diagnosed with rectal gonorrhoea (GC) and chlamydia (CT) have engaged in unprotected receptive anal intercourse. We reviewed the HIV positivity and HIV viral loads (VLs) of men who had rectal GC and CT testing to evaluate potential HIV acquisition and transmission risk. METHODS: Rectal GC and CT testing data for men attending the Maricopa County STD clinic during the period from 1 October 2011 to 30 September 2013 were cross-matched with HIV surveillance data to identify men with HIV coinfection. We examined HIV status, HIV diagnosis date, and the values of VL collected nearest to the date of reported rectal infection. RESULTS: During the 2-year time period, 1591 men were tested for rectal GC and CT. Of the men tested, 506 (31.8%) were positive for GC (13.2%), CT (12.2%) or both (6.4%); 119 (23.5%) of those with rectal GC or CT were coinfected with HIV. Among the 275 men with HIV at the time of rectal testing, 54 (19.6%) had no reported VL; 63 (22.9%) had an undetectable VL (< 20 HIV-1 RNA copies/mL) and 158 (57.4%) had a detectable VL collected within 1 year of rectal diagnosis. Mean VL was higher among HIV and rectal GC/CT coinfected cases compared with men with HIV alone (174 316 vs. 57 717 copies/mL, respectively; P = 0.04). CONCLUSIONS: Approximately one-third of men undergoing rectal testing were positive for GC or CT and one-quarter of men with rectal GC or CT also had HIV infection. Of the HIV-infected men tested for rectal GC or CT, more than half had a detectable VL collected near the time of rectal testing, demonstrating a risk for transmitting HIV.

The association between postprandial urinary C-peptide creatinine ratio and the treatment response to liraglutide: a multi-centre observational study.

AIMS: The response to glucagon-like peptide 1 receptor agonist treatment may be influenced by endogenous ß-cell function. We investigated whether urinary C-peptide creatinine ratio assessed before or during liraglutide treatment was associated with treatment response. METHODS: A single, outpatient urine sample for urinary C-peptide creatinine ratio was collected 2 h after the largest meal of the day among two separate groups: (1) subjects initiating liraglutide (0.6 â†’ 1.2 mg daily) or (2) subjects already treated with liraglutide for 20-32 weeks. The associations between pretreatment and on-treatment urinary C-peptide creatinine ratio and HbA1c change at 32 weeks were assessed using univariate and multivariate analyses (the ratio was logarithm transformed for multivariate analyses). Changes in HbA1c according to pretreatment urinary C-peptide creatinine ratio quartiles are shown. RESULTS: One hundred and sixteen subjects (70 pretreatment, 46 on treatment) with Type 2 diabetes from 10 diabetes centres were studied. In univariate analyses, neither pretreatment nor on-treatment urinary C-peptide creatinine ratio correlated with HbA1c change (Spearman rank correlation coefficient, r = -0.17, P = 0.17 and r = -0.20, P = 0.19, respectively). In multi-linear regression analyses, entering baseline HbA1c and log urinary C-peptide creatinine ratio, pretreatment and on-treatment log urinary C-peptide creatinine ratio became significantly associated with HbA1c change (P = 0.048 and P = 0.040, respectively). Mean (sd) HbA1c changes from baseline in quartiles 1 to 4 of pretreatment urinary C-peptide creatinine ratio were -3 ± 17 mmol/mol (-0.3 ± 1.6%) (P = 0.52), -12 ± 15 mmol/mol (-1.1 ± 1.4%) (P = 0.003), -11 ± 13 mmol/mol (-1.0 ± 1.2%) (P = 0.002) and -12±17 mmol/mol (-1.1±1.6%) (P=0.016), respectively. CONCLUSIONS: Postprandial urinary C-peptide creatinine ratios before and during liraglutide treatment were weakly associated with the glycaemic response to treatment. Low pretreatment urinary C-peptide creatinine ratio may be more useful than higher values by predicting poorer glycaemic response.

Evaluation of point-of-care algorithms to detect diabetes during screening for latent TB infection.

BACKGROUND: Individuals with both diabetes mellitus (DM) and TB infection are at higher risk of progressing to TB disease.OBJECTIVE: To determine DM prevalence in populations at high risk for latent TB infection (LTBI) and to identify the most accurate point-of-care (POC) method for DM screening.METHODS: Adults aged ≥25 years were recruited at health department clinics in Hawaii and Arizona, USA, and screened for LTBI and DM. Screening methods for DM included self-report, random blood glucose (RBG), and POC hemoglobin A1c (HbA1c). Using HbA1c ≥6.5% or self-reported history as the gold standard for DM, we compared test strategies to determine the most accurate method while keeping test costs low.RESULTS: Of 472 participants, 13% had DM and half were unaware of their diagnosis. Limiting HbA1c testing to ages ≥30 years with a RBG level of 120-180 mg/dL helped identify most participants with DM (sensitivity 85%, specificity 99%) at an average test cost of US$2.56 per person compared to US$9.56 per person using HbA1c for all patients.CONCLUSION: Self-report was insufficient to determine DM status because many participants were previously undiagnosed. Using a combination of POC RBG and HbA1c provided an inexpensive option to assess DM status in persons at high risk for LTBI.

Metformin: safety in cardiac patients.

Metformin is a biguanide, insulin sensitiser that reduces blood sugar levels. There are concerns about the risk of lactic acidosis in patients receiving metformin who have procedures requiring iodinated contrast, and in those with renal impairment or heart failure. The data on which these concerns are based are reviewed, with the conclusion that metformin treatment is rarely to blame for lactic acidosis. A generic policy of stopping metformin 48 h before and 48 h after the procedure in all patients is counterintuitive, lacks any evidence base and does not conform to the principles of best practice. In patients with heart failure, although the underlying condition can predispose to lactic acidosis, existing evidence suggests that metformin use is associated with improved outcome rather than increased risk.

Meta-analysis: Helicobacter pylori eradication treatment efficacy in children.

BACKGROUND: Several meta-analyses assessing the efficacy of anti-Helicobacter pylori treatment in adults have been published but a comparable meta-analysis in children is lacking. AIMS: To summarize the efficacy of treatments aimed at eradicating H. pylori in children and to identify sources of variation in treatment efficacy across studies. METHODS: We searched Medline, reference lists from published study reports, and conference proceedings for anti-H. pylori treatment trials in children. Weighted meta-regression models were used to find sources of variation in efficacy. RESULTS: Eighty studies (127 treatment arms) with 4436 children were included. Overall, methodological quality of these studies was poor with small sample sizes and few randomized-controlled trials. The efficacy of therapies varied across treatment arms, treatment duration, method of post-treatment assessment and geographic location. Among the regimens tested, 2-6 weeks of nitroimidazole and amoxicillin, 1-2 weeks of clarithromycin, amoxicillin and a proton pump inhibitor, and 2 weeks of a macrolide, a nitroimidazole and a proton pump inhibitor or bismuth, amoxicillin and metronidazole were the most efficacious in developed countries. CONCLUSIONS: Before worldwide treatment recommendations are given for eradication of H. pylori, additional well-designed randomized placebo-controlled paediatric trials are needed, especially in developing countries where both drug resistance and disease burden is high.

Sex differences in neurotransmission parameters in response to repeated mild restraint stress exposures in intact male, female and ovariectomised female rats.

The present study determined whether a repeated mild restraint stress exposure would differentially alter neuronal activity in male and female rats to gain insights into neurobiological substrates involved in sex differences in stress-induced behavioural responses. In our first set of experiments, we used Western blot analysis to determine whether alterations in several synaptic proteins were elicited by the repeated stress treatment. We found bidirectional changes in synaptophysin levels in female cerebral cortex and hippocampus that diverged between intact and ovariectomised females. There were persistent elevations in spinophilin levels in the male, but not female, hippocampus following the repeated mild restraint stress exposure. By contrast, levels of the NMDA receptor scaffolding protein, PSD-95, were altered only in intact female cerebral cortex and ovariectomised female hippocampus. We next used immunohistochemical evaluation of Fos expression as a marker for neuronal activation. We found significant increases in Fos immunoreactivity in all sex conditions across multiple brain regions in response to the repeated mild stress. Fos protein induction was greatest in the frontal cortex, piriform cortex and amygdala, with the degree of induction varying by sex condition. Fos induction was dramatically higher in amygdala and piriform cortex only in intact females following repeated stress compared to a single restraint stress exposure, suggestive of sensitisation rather than habituation. By contrast, the frontal cortex of intact and ovariectomised females showed habituation to the repeated stressor. Males displayed modest sensitisation in both the frontal cortex and dentate gyrus with no changes in other brain areas. Taken together, these findings show that exposure to a mild repeated stress results in sex differences in synaptic adaptations and patterns of brain activation that likely contribute to observed sex differences in stress-induced behaviours. This approach provides valuable insights into interactions between the hormonal milieu and responses to a repeated mild stress, and further supports the importance of considering hormonal status in treatment of stress-related disorders.

Linezolid-induced optic neuropathy: a mitochondrial disorder?

We report a case of bilateral mitochondrial optic neuropathies secondary to long-term linezolid treatment, show the nature of recovery, review the findings in the literature and propose a potential mitochondrial mechanism for linezolid-induced mitochondrial optic neuropathy. This is an observational case report and literature review with presentation of the clinical course of linezolid mitochondrial optic neuropathies through clinical and psychophysical documentation. Main outcome measures included: visual acuity, funduscopical examinations and peripapillary retinal nerve fibre layer (PRNFL) optical coherence tomography (OCT). A 6-year-old boy presented with bilateral optic neuropathies secondary to 1 year of linezolid treatment for osteomyelitis of the mandible. On presentation, visual acuities were 20/400 in both eyes, with considerable optic disc oedema, hyperaemia and PRNFL swelling confirmed by OCT. 2 weeks after the discontinuation of linezolid, visual acuities returned to 20/25 in both eyes, with reduction in the optic disc oedema, hyperaemia and PRNFL swelling. 3 months after the discontinuation of linezolid treatment, visual acuities were stable at 20/20 in both eyes, with a marked decrease in PRNFL swelling confirmed by OCT, and the development of mild temporal optic disc pallor in both eyes. Doctors should be aware of impairments of vision among patients on long-term linezolid treatment and promptly discontinue treatment to prevent irreversible vision loss. The development and resolution of bilateral optic neuropathies with considerable PRNFL swelling in this patient provide insight into the more general rubric of mitochondrial optic neuropathies.

Everolimus-associated Acute Kidney Injury in Patients with Metastatic Breast Cancer.

Recently, everolimus (Evl) has been introduced in the management of hormone receptor-positive metastatic breast cancer, in combination with aromatase inhibitors. Evl-induced acute kidney injury has hitherto been described in other malignancies, especially renal cell cancer, but only once before in a patient with breast cancer. We describe two cases of Evl-associated nephrotoxicity in patients with breast cancer, one of whom underwent a renal biopsy showing acute tubular necrosis. Both our patients improved after withdrawal of the offending agent and have normal renal functions on follow-up.

Weekly versus Three-Weekly Cisplatin-based Concurrent Chemoradiotherapy as definitive treatment in Head and Neck Cancer- Where do we stand?

PURPOSE: To compare toxicity, compliance, and early response of weekly and 3-weekly cisplatin administration concurrent with radiotherapy as definitive treatment in locally advanced squamous cell carcinoma head and neck. MATERIALS AND METHODS: Patients with histologically proven stage III - IV B head and neck carcinoma presenting from June 2013 to March 2014 were randomly assigned to weekly (35 mg/m2, 6 cycles; arm A) and 3 weekly (100 mg/m2, 3 cycles; arm B) cisplatin with concurrent radiotherapy. RESULTS: 60 patients were randomly assigned to treatment, 30 in each arm. Median follow-up was 8 months (range 4-13). There was no significant difference in grade 3 mucositis between the two arms (75.9% vs 70%, p = 0.20). Grade 3 neutropenia was more frequent in arm B (55.2% vs 26.7%, p = 0.01). Hypomagnesemia was the commonest electrolyte imbalance and it was significantly higher in arm B (60% vs 20%, p = 0.001). Completion rate of scheduled chemotherapy cycles was higher for patients receiving weekly regimen. Response at 3 months was similar for all the patients {Complete Response (66.7% vs 62.1%), p = 0.200}. Our data suggested that there is a reduced need of hospitalization and supportive care measures for patients receiving weekly cisplatin with RT (p = 0.05). CONCLUSIONS: Weekly cisplatin 35 mg/m2 chemotherapy concurrent with radiotherapy is equally effective and less toxic in terms of neutropenia, hypomagnesemia and need for supportive measures than the conventional 3 weekly cisplatin 100 mg/m2 regimen.

EGFR and HER-2/neu Expression in Gallbladder Carcinoma: An Institutional Experience.

BACKGROUND: Gallbladder cancer (GBC) is the most common malignancy of the biliary tract in India. Despite the recent advancement in the understanding of the cancer biology, the disease still remains a therapeutic challenge with poor prognosis and low survival. Surgery is the primary modality of treatment and rest of the modalities are basically adjuvant in nature. This study was performed to evaluate the epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 HER-2/neu) expression in GBC. METHODS: The present study was prospective and done in tertiary super-specialty institute of northern India. This was a pilot study, and at the time of completion, 29 samples were found suitable which were later submitted for EGFR and HER-2/neu evaluation. The sample includes both cases of GBC (n=18) and cholecystitis (n=11) as control. (Table 1) After performing necessary processing, slides were incubated with primary antibody (EGFR) ready to use (RTU) BioGenex, India and HER-2/neu dilution 1:600, Dakopatts (Denmark). Then slides were incubated with secondary antibody (Real Envision Detection Kit, Dakopatts, Denmark). Finally in mounted slides, cell membrane staining was used to assess positivity for EGFR and HER-2/neu. RESULTS: EGFR was positive in 21/29 (72.41%) overall, out of which 14/18 (77.78%) positive in cases and 7/11 (63.64%) positive in control. (Table 1, 3)(Figure 2). For HER-2/neu, it was positive in 21/29 (72.41%) overall, just like in EGFR mentioned above, but positive 12/18 (66.67%) in cases and 9/11 (81.82%) in control respectively. CONCLUSION: Despite the efforts by many investigators, GBC continues to represent a major challenge in oncology. Surgical resection remains the only curative treatment for this disease. The roles of radiation, chemoradiation, and chemotherapy in neoadjuvant and adjuvant settings remained to be defined in prospective studies. With further studies based on molecular understanding and developing new targeted therapies, we will be in better position to manage GBC and increase the survival rate.

Crystallization and molecular packing analysis of barstar crystals.

Barstar, the natural inhibitor of barnase crystallizes in many different crystal forms under almost identical conditions. Although barstar is a monomeric protein, it crystallizes with four molecules in the asymmetric unit in two crystal forms, rhombohedral (space group R3; a = b = 118.0 angstrum; c = 75.5 angstrum) and tetragonal (space group P4; a = b = 105.1 angstrum; c = 36.0 angstrum), which exist simultaneously under identical crystallization conditions. The relation between the four molecules in the asymmetric unit of the crystals belonging to space group P4 can be interpreted in terms of a small distortion in the crystallographic symmetry of the higher symmetry space group P422.

The prognostic role of p53, metallothionein, P-glycoprotein, and MIB-1 in muscle-invasive urothelial transitional cell carcinoma.

Tissue from primary tumors was analyzed for 118 patients with urothelial cancer who subsequently received cisplatin-based chemotherapy. Immunohistochemical staining was performed for nuclear p53 reactivity; for two proposed mediators of drug resistance, metallothionein (MT) and P-glycoprotein; and for the cell proliferation marker MIB-1. For each marker, immunoreactivity was expressed as a percentage of positively staining cells, and overall intensity of staining was graded on a scale from 0 to 3. The product of these two measurements was calculated to generate a percentage-intensity index. Clinical data were obtained independently via retrospective chart review. Chemotherapy regimens containing cisplatin (cisplatin, methotrexate, and vinblastine or methotrexate, vinblastine, doxorubicin, and cisplatin) were administered for metastatic disease (n = 64), for locally advanced disease (n = 45), or as an adjuvant treatment (n = 9). The overall response rate was 56% among 99 evaluable patients, and median survival was 12.7 months. By univariate analysis, Eastern Cooperative Oncology Group performance status (P = 0.0025), tumor grade (P = 0.03), percentage of MT staining (P = 0.01), and percentage-intensity index of MT staining (P = 0.04) were significant predictors of response to chemotherapy. The first three of these were significant in a multivariate model (P = 0.05, 0.04, and 0.04, respectively). By subgroup analysis, the percentage of MT staining predicted for response in metastatic disease (P = 0.03), but not in locally advanced disease (P = 0.28). Only performance status was significantly related to overall survival (P = 0.0001, log-rank test) in the whole cohort. Overexpression of MT in the 64 patients with metastatic disease was associated with a shorter survival (P = 0.04). Expression of p53, P-glycoprotein, and MIB-1 did not predict for survival. In conclusion, overexpression of MT is associated with a poorer outcome from chemotherapy, possibly due to cisplatin resistance.

Management of adrenocortical insufficiency with continuous subcutaneous hydrocortisone infusion: long-term experience in three patients.

UNLABELLED: To assess continuous subcutaneous hydrocortisone infusion (CSHI) in patients with adrenocortical insufficiency (AI) and difficulties with oral replacement. Three patients with AI and frequent hospital admissions attributed to adrenal crises were treated with CSHI, which was delivered via a continuous subcutaneous infusion. All three patients preferred CSHI and remained on it long term, which permitted prolonged follow-up analysis. All three patients reported symptomatic improvement, and in two cases, reduced hospital admission rates and inpatient stay lengths were observed. The cost of hospital admissions and overall treatment was reduced in all cases. CSHI offers a practical and acceptable alternative to oral replacement in a subset of patients with AI. The cost of initiating and maintaining the pump is offset in the long term by reduced frequency and duration of emergency admissions. CSHI can therefore be considered in a select group of patients who are resistant to treatment with conventional oral glucocorticoids. LEARNING POINTS: Continuous subcutaneous infusion of cortisol is a viable alternative in patients unable to take oral steroids.Patient acceptability was high, with three out of three patients preferring to remain on pump treatment.Hospital admissions were reduced in response to pump therapy, which compensated for the increased treatment cost.The daily dosage of hydrocortisone can be reduced by using pump therapy.

pH dependence of the stability of barstar to chemical and thermal denaturation.

Equilibrium unfolding of barstar with guanidine hydrochloride (GdnHCl) and urea as denaturants as well as thermal unfolding have been carried out as a function of pH using fluorescence, far-UV and near-UV CD, and absorbance as probes. Both GdnHCl-induced and urea-induced denaturation studies at pH 7 show that barstar unfolds through a two-state F<->U mechanism and yields identical values for delta GU, the free energy difference between the fully folded (F) and unfolded (U) forms, of 5.0 +/- 0.5 kcal.mol-1 at 25 degrees C. Thermal denaturation of barstar also follows a two-state F<->U unfolding transition at pH 7, and the value of delta GU at 25 degrees C is similar to that obtained from chemical denaturation. The pH dependence of denaturation by GdnHCl is complex. The Cm value (midpoint of the unfolding transition) has been used as an index for stability in the pH range 2-10, because barstar does not unfold through a two-state transition on denaturation by GdnHCl at all pH values studied. Stability is maximum at pH 2-3, where barstar exists in a molten globule-like form that forms a large soluble oligomer. The stability decreases with an increase in pH to 5, the isoelectric pH of the protein. Above pH 5, the stability increases as the pH is raised to 7. Above pH 8, it again decreases as the pH is raised to 10. The decrease in stability from pH 7 to 5 in wild-type (wt) barstar, which is shown to be characterized by an apparent pKa of 6.2 +/- 0.2, is not observed in H17Q, a His 17-->Gln 17 mutant form of barstar. This decrease in stability has therefore been correlated with the protonation of His 17 in barstar. The decrease in stability beyond pH 8 in wt barstar, which is characterized by an apparent pKa of 9.2 +/- 0.2, is not detected in BSCCAA, the Cys 40 Cys 82-->Ala 40 Ala 82 double mutant form of barstar. Thus, this decrease in stability has been correlated with the deprotonation of at least one of the two cysteines present in wt barstar. The increase in stability from pH 5 to 3 is characterized by an apparent pKa of 4.6 +/- 0.2 for wt barstar and BSCCAA, which is similar to the apparent pKa that characterizes the structural transition leading to the formation of the A form. The use of Cm as an index of stability has been supported by thermal denaturation studies.(ABSTRACT TRUNCATED AT 400 WORDS)

Association of partially-folded intermediates of staphylococcal nuclease induces structure and stability.

Staphylococcal nuclease forms three different partially-folded intermediates at low pH in the presence of low to moderate concentration of anions, differing in the amount of secondary structure, globularity, stability, and compactness. Although these intermediates are monomeric at low protein concentration (< or =0.25 mg/mL), increasing concentrations of protein result in the formation of dimers and soluble oligomers, ultimately leading to larger insoluble aggregates. Unexpectedly, increasing protein concentration not only led to association, but also to increased structure of the intermediates. The secondary structure, stability, and globularity of the two less-ordered partially-folded intermediates (A1 and A2) were substantially increased upon association, suggesting that aggregation induces structure. An excellent correlation was found between degree of association and amount of structure measured by different techniques, including circular dichroism, fluorescence, Fourier transform infrared spectroscopy (FTIR), and small-angle X-ray scattering. The associated states were also substantially more stable toward urea denaturation than the monomeric forms. A mechanism is proposed, in which the observed association of monomeric intermediates involves intermolecular interactions which correspond to those found intramolecularly in normal folding to the native state.