Serum and urine galactose deficient-IgA1 as alternative biomarkers in the management of IgA nephropathy.

INTRODUCTION: Immunoglobulin A (IgA) nephropathy (IgAN) results from abnormal accumulation of immune complexes containing galactose deficient IgA1 (Gd-IgA1) in the kidneys. About 40% of patients develop end-stage kidney disease within 20 years of renal biopsy. At present, the diagnosis and risk stratification of patients (using the international IgAN risk prediction tool) rely on renal biopsy, which is an invasive procedure. Also, treatment decisions are still dependent on proteinuria, which is not specific for IgA nephropathy. We discussed the role of serum and urine Gd- IgA1 in the diagnosis of IgAN, its association with disease progression and changes with treatment in patients with IgA nephropathy. MATERIALS AND METHODS: A systematic search of PubMed and Scopus databases was done to identify the articles that are relevant to the topic including systematic reviews and original articles. RESULTS: Several studies showed that both serum and urine Gd-IgA1 differentiate IgA nephropathy patients from healthy people and other glomerulonephropathies. Thus, it is useful as a less invasive diagnostic biomarker, although detection methods varied between studies with different sensitivities. There are various reports of its use as a prognostic parameter. Evidence is emerging for its use as a monitoring parameter for treatment. CONCLUSION: Galactose deficient IgA1 is a promising biomarker in the management of IgA nephropathy, although a more robust and standardised means of estimation is required.

Hedgehog signalling molecule, SMO is a poor prognostic marker in bladder cancer.

INTRODUCTION: Hedgehog (HH) pathway is an important signalling cascade for growth and patterning during embryonic development. Constitutive activation of Hedgehog pathway can be found in various types of malignancies including medulloblastoma, basal cell carcinoma, gastrointestinal, breast, pancreatic, prostate cancer and leukaemia. Little is known about the expression and role of Hedgehog signalling in bladder cancer. MATERIALS AND METHODS: The purpose of this study was to investigate the immunohistochemical expression of SMO in 112 bladder cancer cases and determine their association with demographic and clinicopathological parameters. Bladder cancer tissues were obtained from the Hospital Kuala Lumpur. RESULTS: SMO was expressed in the cytoplasm of all cases of bladder cancer. 6 cases (5.4%) showed low expression, while 106 cases (94.6%) showed high expression. Positive expression of SMO protein was correlated with a few variables which include grade and stage of tumour, lymph node metastasis and distant metastasis. SMO expression showed statistically significant association with higher grade (p=0.001) and higher stage (p=0.042) of bladder cancer. SMO expression also showed borderline association with lymph node metastasis (p=0.056). CONCLUSION: These findings indicate that SMO expression may be a poor prognostic marker in bladder cancer.

Focal Segmental Membranoproliferative Glomerulonephritis: A Histological Variant of Denys-Drash Syndrome.

Introduction: Denys-Drash Syndrome (DDS) consists of a triad of pseudohermaphroditism, Wilms'tumor and nephropathy. This condition may manifest as a complete triad or in an incomplete form; with either one or a combination of the above features. The characteristic glomerular abnormality in DDS is diffuse mesangial sclerosis (DMS).Case report: We report two cases of DDS with focal membranoproliferative glomerulonephritis (MPGN). Both of our cases were males with ambiguous genitalia. They had a similar heterozygous germline mutation in exon 9 of WT1, c.1180C>T, p.R394W; a known mutation hotspot for DDS. Case 1 had nephropathy at the age of 4 years and Case 2 at 2.5 years with different rates of progression to end-stage renal failure. Conclusion: Our findings, in combination with other reports, illustrate the clinicopathological heterogeneity of DDS. There are no universal recommendations for optimal management of patients with DDS due to the inability to accurately predict affected individuals' progress.

Immunohistochemical expression of NANOG in urothelial carcinoma of the bladder.

Urothelial carcinoma is a common malignant neoplasm that has a poor prognosis and a high frequency of recurrence and metastasis. Constant disease surveillance with periodic and long term cystoscopy examination is necessary for management of the disease. However, the monitoring and therapy regimen is expensive, incurring a massive burden to patients and the government. Therefore, the development of specific biomarkers for urothelial carcinoma at an early stage and recurrence detection becomes a priority. Homeobox genes are a family of genes that are involved in tumourigenesis. They might be potential prognostic markers for urothelial carcinoma. The study investigated the expression pattern of NANOG which is one of a homeobox gene in different stages and grades of urothelial carcinoma. NANOG expressions were also correlated with patient demographic factors and clinicopathological parameters. The expression of NANOG in 100 formalin-fixed paraffin-embedded urothelial carcinoma tissues was determined by immunohistochemistry. Immunohistochemistry showed positive expression of NANOG in all specimens with detection in the cytoplasm, nuclei and the nuclear membrane of the cancer cells. The immunohistochemical expression of NANOG increased across stages and grades of the tumour. The expression of NANOG was not significantly associated with demographic factors; gender (p = 0.376), race (p = 0.718) and age (p = 0.058) as well as with most of the clinicopathological parameters; pathological stage (p = 0.144), grade (p = 0.625), lymph node involvement (p = 0.174) and distant metastasis (p = 0.228). However, NANOG expression showed significant correlation with tumour invasion (p = 0.019). We concluded that NANOG might be a potential biomarker for early diagnosis of urothelial carcinoma of the bladder.

Gender and age differences in walking for transport and recreation: Are the relationships the same in all neighborhoods?

INTRODUCTION: Walking as regular physical activity (PA) is central to healthy aging, and environments influence walking. Multilevel neighborhood-based studies that only report average (fixed-effect) walking differences for gender and age implicitly assume that neighborhood environments influence the walking behavior of men and women, and younger and older persons, similarly. This study tests this assumption by examining whether gender and age differences in walking for transport (WfT) and walking for recreation (WfR) are similar or different across neighborhoods. METHODS: This paper used data from the HABITAT multilevel study, with 7,866 participants aged 42-68 years in 2009 living in 200 neighborhoods in Brisbane, Australia. Respondents reported minutes spent WfT and WfR in the previous week, categorized as: none (0 mins), low (1-59mins), moderate (60-149mins) and high (≥ 150 mins). Multilevel multinomial logistic models were used to estimate average differences in walking by gender and age, followed by random coefficients to examine neighborhood variation in these individual-level relationships. RESULTS: On average, women were more likely to engage in WfR at moderate and high levels (no gender differences found in WfT); and older persons were less likely to do WfT and more likely to do high levels of WfR. These average (Brisbane-wide) relationships varied significantly across neighborhoods. CONCLUSION: Relationships between gender and walking, and age and walking, are not the same in all neighborhoods, (i.e. the Brisbane average conceals important information) suggesting that neighborhood-level factors differentially influence the walking behaviors of men and women and younger and older persons. Identifying these factors should be a priority for future research.

Serum HER2 extracellular domain in metastatic breast cancer patients treated with weekly trastuzumab and paclitaxel: association with HER2 status by immunohistochemistry and fluorescence in situ hybridization and with response rate.

PURPOSE: We explored the relationship between circulating HER2 extracellular domain (ECD) and tissue HER2 status as determined by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). We also examined its predictive value in a cohort of metastatic breast cancer patients treated with weekly trastuzumab and paclitaxel. METHODS: Eligible patients had pre- and post-treatment stored serum specimens and were treated on a previously reported phase II trial. Retrospective analysis evaluated: the association between pretreatment serum HER2 ECD and tissue HER2 status by IHC and FISH; and the association between change in serum HER2 ECD after 12 weeks of therapy and response proportion. RESULTS: Stored serum samples were available for 55/95 (58%) patients. Statistically significant associations were found between HER2 status as assessed by IHC and FISH, and baseline serum HER2 ECD level. Patients whose ECD normalized after 12 weeks of therapy had a higher response proportion compared with patients with persistently high ECD levels (68% versus 15%, P=0.005). A relative decline of over 55% from baseline HER2 ECD predicted response to therapy. CONCLUSION: A statistically significant association was observed between pretreatment serum HER2 ECD and tissue HER2 status as assessed by IHC and FISH. A decrease in serum HER2 ECD level was a significant predictor of response to trastuzumab-based therapy.

Shoulder function and patient well-being after various types of neck dissections.

Preserving the accessory nerve results in a better outcome of the shoulder function after neck dissection. However, little is known about the impact of preserving a cervical contribution to the accessory nerve. This study describes the shoulder function after different types of neck dissections, with the emphasis on the significance of the cervical contribution to the accessory nerve. Fifty-nine patients who underwent neck dissections of various types were included. Thirty-eight patients underwent unilateral radical or modified radical neck dissections, and 21 patients underwent bilateral neck dissections. All the patients were assessed subjectively and objectively, using a questionnaire and an inclinometer. Radical neck dissections inflicted significantly more morbidity than modified radical neck dissections. Preserving a cervical contribution to the accessory nerve did not decrease pain complaints or functional impairment. However, there might be some improvement in range of motion, especially exorotation and anteflexion. Preserving the accessory nerve has a positive influence on shoulder function and complaints. Preserving a cervical contribution does not decrease morbidity significantly.

Toxoplasmosis and habitual abortion.

We set out to determine the role of toxoplasmosis, detected by serological tests, in habitual abortion. A total of 280 pregnant women aged 15-46 years with parity ranged from 0-9 were studied prospectively between January 2000 and May 2001 at King Hussein Medical Center. Analyses for IgG and IgM anti-toxoplasma were carried out using indirect fluorescent antibody assay (IFAT) and enzyme-linked immunosorbent assay (ELISA). Titres of the order of 1:16-1:2048 were considered positive. According to the results, women were divided into two groups; seropositive (n=132) and seronegative (n=148). One hundred and thirty-two (47.1%) pregnant women showed seropositivity to IgG anti-toxoplasma; of them, two (1.5%) developed IgM anti-toxoplasma during the second trimester. A statistically significant increase in the rate of seropositivity to toxoplasma with increasing age and parity was found (P<0.05). There was no significant difference in the rate of habitual abortion between seropositive and seronegative women. The seropositivity was higher among women living in rural areas (P<0.02), who are using rainwater to drink (P<0.02), ingesting undercooked meat (P<0.001) and who have contact with soil (P<0.02). Toxoplasma antibodies detected by positive serological tests tend to be higher with increased age and parity. It seems that they have no role in habitual abortion.

Low-level lead exposure and blood lead levels in children: a cross-sectional survey.

The authors studied 53 girls (44.5%) and 66 (55.5%) boys in Karachi, Pakistan, to determine their blood lead levels. The association between blood lead levels/water lead levels and the possible risk factors and symptoms associated with lead toxicity was explored. The mean lead level for the entire group was 7.9 microg/dl (standard deviation = 4.5 microg/dl). Thirty (25.2%) of the children had lead levels that exceeded 10 microg/dl; 12 (10.0%) of these had lead levels that exceeded 15 microg/dl. Thirteen (20.9%) of the children under the age of 6 yr (n = 62) had lead levels greater than 10 microg/dl, and 6 (9.6%) had levels in excess of 15 microg/dl. The authors found no association (p > .05) between high lead levels in water and blood lead levels in children. Mean blood lead levels were highest in the group of children exposed to various risk factors for lead absorption (e.g., exposure to paint, remodeling, and renovation; use of lead utensils; pica). There was a significant association between a history of exposure to paint/renovation activities and a history of pica. High blood lead levels in the children in Karachi stress the urgency for actions that control lead pollution. Screening programs should be instituted by the state. Individuals must become aware of lead's toxicity, and they must avoid substances that contain lead.

Reflex testing II: evaluation of an algorithm for use of cardiac markers in the assessment of emergency department patients with chest pain.

A reflex algorithm was developed and evaluated for the use of serum cardiac markers for the diagnosis and rule out of acute myocardial infarction (AMI), and risk stratification of unstable angina patients for those who present to emergency departments (ED) with chest pain. The process begins with testing of total CK and myoglobin at admission. Based on these results, the algorithm determines the need for subsequent testing for the CK-MB isoenzyme and cardiac troponin I (cTnI). The algorithm also directs the need for further blood collection and cardiac marker testing at 4, 8, and 12 h after presentation. A total of eleven stopping points were identified. For some of these stopping points, the algorithm concluded that further blood collections and testing was unnecessary and redundant. The algorithm was retrospectively evaluated on 101 non-consecutive chest pain patients who presented to the EDs at three hospitals. For the AMI group (n=34), six of nine possible different stopping points were reached: 64.7% of cases were diagnosed with the first sample at admission, an additional 32.3% after 4 h, and 2.9% at 8 h. The 12-h sample was not necessary for any of the AMI patients. For the non-AMI group (n=67), most reached the stopping point of no cardiac injury or risk. There were five unstable angina patients who had minor myocardial damage on the basis of a marginally increased cTnI. Of these, one patient subsequently suffered AMI, and three others required angioplasty or bypass surgery. Compared to performing four tests on all patient samples, the reflex algorithm would have reduced the number of necessary tests from 442 to 130 (71% reduction) for AMI patients, and 871 to 469 (46% reduction) for non-AMI patients, if prospectively implemented.

[Otosclerosis surgery with CO2 laser]. / Cirugía de la otoesclerosis con láser CO2.

Since Shea described the stapedectorny, many physicians have developed modifications of the technique, prosthesis or instrumentation. We have adopted one of the most recent advances to the treatment of otosclerosis, CO2 laser. Interventions were carried out in 20 consecutive ears using CO2 laser. Otosclerosis with type I, II and III foci were diagnosed and stapedectomies were reviewed. Stapedectomy was performed with a Causse prosthesis. We describe the technique and the laser power and application time used. The preoperative and auditory results at 1 month are reported. The advantages of opening otosclerosis with CO2 laser as the first or review procedure are analyzed.

Method for enhancing retention in complete denture bases.

STATEMENT OF PROBLEM: Physical retention of a denture depends on adhesion of a salivary film to its fitting surface and supporting tissues. Adhesion is influenced by many factors. PURPOSE: This study examined the effect of the texture of the fitting surface on denture retention. MATERIAL AND METHODS: According to the method developed, mucosal-supported palatal bases were made for 10 dentate subjects who had no obvious oral disease, history of allergy to dental materials, or palatal mucosal undercuts. A specially designed strain gauge force transducer and a pen-chart recorder were used to record forces that were needed to dislodge the bases from the palate. Forces were recorded 30 times (10/day) for each subject. Similar forces were recorded after the fitting surface of each base was uniformly air-particle abraded under 4 kg/cm2 of pressure for 1 minute with 50-microm grit alumina dust. Data were analyzed with a 2-sample Student t test and a paired t test. RESULTS: Substantial variations for all measurements were observed both between and within subjects. Notwithstanding, these variations, the mean retention of the bases significantly (P <.05) increased by more than 2-fold when the fitting surface was air-particle abraded. These force values were similar to those recorded previously for well-fitting mucosal-supported palatal bases in a separate group of dentate subjects. CONCLUSIONS: While reinforcing the importance of good adaptation of the denture to the tissues, these findings also suggest that an additional improvement in denture retention can be accomplished after the fitting surface is air-particle abraded.

Role of serum angiotensin converting enzyme in sarcoidosis.

This study was conducted to determine the role of Serum Angiotensin Converting Enzyme (SACE) as a marker in the differential diagnosis of pulmonary diseases and prognosis of sarcoidosis. A retrospective analysis of 113 medical records of patients at The Aga Khan University Hospital, with laboratory investigation for SACE was performed. Among 113 patients, 51 cases were found to have sarcoidosis, 44 of them had SACE levels greater than 52 IU/L (mean ACE 104.44). SACE levels were also found elevated in other clinical conditions like tuberculosis (mean 58.64 IU/L), but the enzyme level were less (p 0.04) than those found in sarcoidosis (mean (92.97 IU/L). SACE activity was found to be considerably lower in other chronic lung diseases such as, fibrosing alveolitis (mean 43.98 IU/L), interstitial lung disease (mean 42.11 IU/L) and chronic obstructive lung disease (mean 40.85 IU/L). Twenty patients of sarcoidosis, who received steroid treatment subsequently showed a decline in the SACE levels. SACE is a useful marker in differential diagnosis as 37.2% cases of sarcoidosis compared to only 9.09% of tuberculosis had SACE levels greater than 100 IU/L. In addition, our data also suggest that serum ACE is useful for the diagnosis as well as monitoring prognosis in sarcoidosis.

Diagnosis of genetic defects by chromosomal analysis.

Of 901 karyotypes performed over a period of 4 years, genetic anomalies were detected in 162 cases. Down's syndrome (trisomy 21) was the most common (168.8%) genetic disorder followed by Turner's syndrome, Philadelphia chromosome, Klinefelter's syndrome, Edward's syndrome (trisomy 18) and Patau's syndrome (trisomy 13). All the three trisomies were detected very early in life. Mean age at the time of diagnosis for Turner's syndrome was 13.3 years, allowing a timely hormone replacement therapy to improve secondary sexual characters. Patients with Klinefelter's syndrome were diagnosed late (mean age 23.6 years), which greatly reduced their chances of an effective therapy to improve the clinical and social outcome.

An investigation into the effect of denture fixatives in increasing incisal biting forces with maxillary complete dentures.

The effect of denture fixatives on incisal biting forces was investigated in five subjects wearing maxillary complete dentures. Forces required to displace new and old dentures with and without fixatives were measured up to six hours post-insertion using a force transducer. The results showed that incisal forces were greater with new dentures than with old dentures when only saliva was present. After the application of fixative to the old dentures, the difference from the new dentures without fixative became insignificant. It is concluded that the improvement in retention of complete dentures with the use of fixatives allowed patients to increase their incisal biting capability.

Some clinical investigations on retention forces of maxillary complete dentures with the use of denture fixatives.

The aim of this study was to evaluate objectively and clinically the relative improvement in retention forces of complete dentures with the use of three proprietary denture fixatives. Using the UCL Retentiometer, retention forces of ill-fitting maxillary complete dentures were measured with saliva alone as the interface medium and with the use of the three test denture fixatives in edentulous subjects over a period of 6 h. Results showed that forces with saliva alone were consistently and significantly lower when compared with the forces measured at any test period with the use of the test fixatives. A sixfold improvement in forces from the salivary baseline values was observed when these materials were used. However, the test fixatives showed substantial variations in retention forces depending on the physical formulation of the material. Whilst the powder variety was effective immediately after its application, retention forces with the liquid and paste materials did not peak until 3 h post-insertion. Measurements at 6 h post-insertion stage showed that the powder fixative was the least effective of the three materials. This was mainly attributed to the rapid washing of the active ingredients by saliva because of the absence of oily base present in the paste and liquid materials.

Inhibition of a reductive function of the plasma membrane by bacitracin and antibodies against protein disulfide-isomerase.

Evidence had been provided that a disulfide-linked [125I]iodotyramine/poly(D-lysine) conjugate was reductively cleaved when bound nonspecifically to the surface of Chinese hamster ovary (CHO) cells and that this cleavage was abolished by membrane-impermeant sulfhydryl blockers. The same blockers were subsequently found to inhibit the cytotoxicity of diphtheria toxin, a disulfide-linked heterodimer that binds to a specific surface receptor and must undergo chain separation to exert its cytotoxicity. This suggested that the disulfides of both macromolecules might be cleaved by a thiol-disulfide interchange reaction, possibly mediated by protein disulfide-isomerase (PDI, EC 5.3.4.1). We tested whether inhibitors of PDI--in particular, bacitracin and anti-PDI antibodies--might mimic the two effects of sulfhydryl blockers. Both bacitracin and anti-PDI antibodies were effective in inhibiting both reductive processes. This strongly suggests that the disulfide cleavage in the two membrane-bound macromolecules is mediated by PDI and that this enzyme, besides its known retention in the endoplasmic reticulum, must also be exposed at the plasma membrane. This paper points to other potentially important disulfide reductions that might be catalyzed by surface-associated PDI. It thereby broadens the known functions of an enzyme already known for its multifunctional properties.

Cell surface sulfhydryls are required for the cytotoxicity of diphtheria toxin but not of ricin in Chinese hamster ovary cells.

A previous study on cleavage of disulfide bonds in endocytosed model compounds had shown that an initial phase of cleavage was totally inhibited by membrane-impermeant sulfhydryl inhibitors and thus was mediated by cell surface sulfhydryls (Feener, E. P., Shen, W.-C., and Ryser, H. J.-P. (1990) J. Biol. Chem. 265, 18780-18785). This paper uses the same inhibitors (5,5'-dithiobis(2-nitrobenzoic acid) and p-chloromercuriphenylsulfonic acid) to examine the role of surface sulfhydryls in the cytotoxicity of diphtheria toxin (DT). Since the interchain disulfide of endocytosed DT must be cleaved prior to translocation of chain A from endosomes to cytoplasm, it was postulated that surface sulfhydryls might mediate the cleavage of that disulfide bond as well. Both sulfhydryl blockers did indeed markedly inhibit DT cytotoxicity. This effect was not due to inactivation of unbound DT, inhibition of receptor-mediated endocytosis, or impairment of acidification of endosomes. We conclude that cell surface sulfhydryls susceptible to blockage by 5,5'-dithiobis(2-nitro-benzoic acid) and p-chloromercuriphenylsulfonic acid are required for the cytotoxicity of DT and, most likely, for the reductive cleavage of DT's interchain disulfides. Ricin cytotoxicity was not decreased; this is consistent with the view that ricin reaches the cytoplasm from a late endocytic structure and with the finding that endocytosed disulfides are also cleaved in a cell fraction containing elements of the Golgi apparatus (Feener, E. P., Shen, W.-C., and Ryser, H. J.-P. (1990) J. Biol. Chem. 265, 18780-18785).

Isolation of variants of Chinese hamster ovary cells with abnormally low levels of GSH: decreased ability to cleave endocytosed disulfide bonds.

Mutants of Chinese hamster ovary cells were selected for resistance to a 3 hour exposure to 4 microgram/ml N-methyl-N'-nitro-N-nitrosoguanidine and tested for glutathione (GSH) levels. Six of eight clones that survived the initial treatment had reduced GSH levels ranging from 26 to 61% of wild-type values. These eight cell lines were tested for their susceptibility to a drug conjugate in which methotrexate (MTX) is disulfide-linked to poly(D-lysine) (MTX-SS-PDL) to test their capacity to cleave the endocytosed disulfide bond and release free MTX from this otherwise undegradable drug conjugate. We had shown that wild-type cells were killed by approximately 1 x 10(-7) M MTX given as free drug, as MTX-poly(L-lysine) or as MTX-SS-PDL, but were not affected by MTX-poly(D-lysine). All six lines with abnormally low levels of GSH were resistant to MTX-SS-PDL. The variants with the lowest levels of GSH (MNR-5 and MNR-10) were tested further and showed near-normal sensitivity to MTX and MTX poly(L-lysine). As expected, both lines were hypersensitive to melphalan. They were, however, normally sensitive to diphtheria toxin and ricin, indicating that some cleavage of the interchain disulfides in these protein toxins occurs even when cellular GSH is abnormally low. The lesser GSH requirement for toxin activation may be due to their extraordinary potency.