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[Coloration of cytologic thick sections containing biomaterials with silver methenamine. Usefulness for scanning electron microscopy].

UNLABELLED: Prostheses implanted in hard tissues cannot be processed for electron microscopic examination or microanalysis in the same way as those in other tissues. For these reasons, we have developed a method allowing light and electron microscopic studies as well as microanalysis of the interface between bone and biomaterials after the sections glycoproteins have been stained with silver methenamine. Silver can be evidenced by SEM in back scattered mode. MATERIALS AND METHODS: Tranverse sections of a human femur containing an HA-coated prostheses were obtained with a diamond saw and ground to a thickness of 50-100 microns. The sections were stained in a microwave oven using a 1% silver methenamine solution. They have been examined by back-scatter SEM operated at 25 KV. EDS has been performed on cellular inclusions and extracellular bone matrix. RESULTS: Type I and III collagen fibers, and reticulin fibers were stained. The mineralized matrix was heavily colored. At the cell level, the nuclear and cytoplasm membranes, the chromatin and ribosomes were shown. The characteristic peaks of the Ag spectrum are distinct from those of the elements used in orthopaedic biomaterials and did not impair their identification.

Animals↗

The generation of formaldehyde from methenamine. Effect of urinary flow and residual volume.

The antimicrobial effect of methenamine in urine is dependent upon: (i) the rate of generation of formaldehyde; (ii) the concentration of formaldehyde achieved; and (iii) the duration of bacterial exposure to formaldehyde. Studies utilizing an in vitro model which simulates the dynamics of the urinary tract showed that bacteriostatic concentrations of formaldehyde (greater than or equal to 25 mug per ml) were obtained from attainable concentrations of methenamine at urine pH 5.7 to 5.85. Reduced urinary flow rates and large residual volumes increased formaldehyde concentrations and would appear to provide prolonged bacterial exposure to the formaldehyde.

Formaldehyde↗

[Detection of Pneumocystis carinii in clinical samples by the polymerase chain reaction. Comparison with the methenamine silver technique].

BACKGROUND: A comparison of polymerase chain reaction (PCR) with methenamine silver stain was performed on clinical specimens for diagnosis of Pneumocystis carinii pneumonia in immunosuppressed patients. METHODS: 25 clinical samples (10 induced sputa, 12 transthoracic needle aspirations and 2 bronchoalveolar lavages) from 17 patients with AIDS and respiratory symptoms were studied. We performed DNA amplification by using 2 primers that amplify the mitochondrial large rDNA sequence of P. carinii. RESULTS: The sensitivity and specificity of the PCR technique were 92.8% and 100% respectively, with a positive predictive value of 100% and negative predictive value of 92.3%. The sensitivity of the silver stain were 65%, specificity 100%, positive predictive value 100% and negative predictive value 63%. CONCLUSION: The PCR technique is more sensitive than the methenamine silver stain for detecting Pneumocystis carinii in respiratory specimens from patients with pneumonia. Consequently it could be a very good alternative to classical methods in diagnosis of P. carinii infections.

AIDS-Related Opportunistic Infections↗

Utility of reflex Gomori methenamine silver staining for Pneumocystis jirovecii on bronchoalveolar lavage cytologic specimens: a review.

Pneumocystis jiroveci (Pj; formerly Pneumocystis carinii) is an opportunistic pathogen causing life-threatening pneumonia (Pneumocystis pneumonia) in immunosuppressed individuals. Its diagnosis is dependent on identification in bronchoalveolar lavage (BAL) specimens. Gomori's methenamine silver nitrate (GMS) stain has been advocated to highlight the organisms in BAL specimens. This study was performed to determine the utility of reflex GMS staining on all BAL specimens for identifying Pj.All BAL specimens from years 2000 to 2004 were processed as cytospins and stained with Papanicolaou (Pap) and GMS stains. A total of 2,984 BAL specimens were identified. A total of 116 (3.9% of total BAL) BAL specimens were diagnostic of Pj. The diagnostic specimens were grouped as follows: 103 (88.8% of total positive cases) Pj identified with both Pap and GMS staining; 11 (9.5% of total positive cases) Pj identified only with Pap staining; and 2 (1.7% of total positive cases) Pj identified only with GMS staining. In conclusion, the prevalence of Pj in BAL specimens is 3.9%, which can be attributed to improved management of immunocompromised patients. Performing reflex GMS staining on all BAL specimens does not improve the diagnostic identification of Pj since the majority (98.3%) of diagnoses can be rendered on Pap stained slides. A cost analysis for GMS staining on 2,879 GMS-negative BAL specimens was estimated at $143,950. Thus, from diagnostic and cost benefit perspectives, GMS staining can be recommended only on cases where Pap stain is negative, and the clinical presentation is consistent with Pneumocystis pneumonia.

Adolescent↗

Cytology of treated and minimal Pneumocystis carinii pneumonia and a pitfall of the Grocott methenamine silver stain.

To clarify the role of foamy alveolar casts (FACs) in the diagnosis of Pneumocystis carinii pneumonia (PCP), we retrospectively reviewed Papanicolaou (Pap)- and Grocott methenamine silver (GMS)-stained slides from 205 bronchial specimens submitted for suspected opportunistic lung infection. FACs containing sporozoites were seen in 86 cases, all with positive GMS. FACs were absent in 119 cases with negative GMS. In patients previously treated for PCP, Pap showed FAC-like material which lacked sporozoites. GMS demonstrated clumped degenerated Pneumocystis carinii (PC) cysts. In 17 GMS-positive cases there were no FACs on Pap due to sampling error and low burden of organisms. Morphologic features of macrophages in these cases can suggest the presence of PC. When FACs are lacking, a special stain is required. When GMS is used, the control must contain PC to avoid a false negative GMS. The inconsistent uptake of GMS by PC is a pitfall which is distinct from sampling error and which to our knowledge has not been previously reported.

Bronchi↗

GLC determination of methenamine in tablets.

A rapid and sensitive GLC method was developed for the quantitative determination of methenamine in tablets. The method was shown to possess several advantages over the official NF assay. After dissolution of the whole tablet in absolute ethanol and addition of an internal standard (pentylenetetrazol), an aliquot was injected into the gas chromatograph for analysis. The sample was chromatographed using a stainless steel column packed with 10% OV-17 on Chromosorb W-HP. Quantitation was achieved by measuring peak heights. The simplicity, directness, extreme rapidity, and accuracy of the method represents an improvement over the official method and the other proposed assays.

Chromatography, Gas↗

Distinctive, rapid, and easy labeling of diffuse plaques in the Alzheimer brains by a new methenamine silver stain.

We have developed a new methenamine silver (MS) stain for detecting diffuse plaques distinctively on paraffin-embedded tissue sections of Alzheimer-type dementia, Down's syndrome, and mentally normal aged brains. This rapid and easy method selectively labels amyloid-related component of senile plaques, but not of kuru plaques found in Gerstmann-Sträussler syndrome. Our MS stain shows almost the same staining pattern as that of the beta protein immunostaining with formic acid pretreatment. Therefore, new MS stain is appropriate to routine or screening studies for senile plaques including diffuse plaques.

Alzheimer Disease↗

Ultrastructural localization of argyrophilic substances in diffuse plaques of Alzheimer-type dementia demonstrated by methenamine silver staining.

The ultrastructure of argyrophilic substances in diffuse plaques of an Alzheimer-type dementia brain was examined using methenamine silver (MS) electron microscopy with the pre-embedding method. Electron-dense substances, which were sparse aggregations of bundle-like structures with silver granules, were disseminated in the diffuse plaques. Comparison of serial MS and routine ultrathin sections revealed that diffuse plaques usually show scattered bundles of amyloid fibrils or amorphous materials, or both, between indistinct cell process membranes. Some of these processes were identified as astrocytic or dendritic in origin. A few degenerative neurites were frequently noted in large silver-positive areas but never in small areas. These findings suggest that the appearance of amyloid fibrils and amorphous materials between certain cell processes is a very early morphological change in the process of senile plaque formation.

Aged↗

Standardization of various applications of methacrylate embedding and silver methenamine for light and electron microscopy immunocytochemistry.

The use of butyl-methyl-methacrylate embedding and the application of the silver methenamine (SM) method as a poststaining of the immunoperoxidase-DAB (IP) procedure led to the standardization of several useful methods for the visualization of tissue antigens at the light and electron microscope level. These procedures included: 1) Standardization of the actual methacrylate embedding; 2) The IP-SM method with and without periodic acid oxidation, which provided 100% intensification of the IP staining; 3) The IP-SM method made it possible to stain semithin sections (0.5 micron), and this in turn, permitted a) clear visualization under the light microscope of the intracellular distribution of antigens and, b) staining, in several adjacent sections, of roughly the same cytoplasmic region of the same cell with different primary antisera; 4) a double immunostaining whereby the first antigen in the sequence was revealed by the IP-SM method and the second by the IP procedure; 5) standardization of the IP and the IP-SM methods for post-embedding staining of ultrathin methacrylate sections. The combined application of methacrylate embedding and the IP-SM, and the use of an appropriate fixative, resulted in an ultrastructural immunocytochemical procedure characterized by a good immunoreactivity of the tissue sections, a strong and selective immunoreaction and a well preserved ultrastructure.

Acrylates↗

Effect of short-term high-dose treatment with methenamine hippurate on urinary infection in geriatric patients with an indwelling catheter. IV. Clinical evaluation.

An evaluation has been made of the clinical and laboratory effects of short-term (34 days), high-dose (2g x 3 daily) treatment with methenamine hippurate (MH) of 14 geriatric patients with an indwelling catheter and clinical features of urinary tract infection. During MH treatment the number of catheter changes was halved, each catheter remaining in situ for an average of 12.0 days as compared to 6.2 days in the pre-treatment control period and 5.2 days in the post-treatment control period; the difference is significant (p = 0.008; Friedman two-way analysis of variance). Urine pH was reduced (pH 7.0--6.5--7.0; p = 0.01) and the standard bicarbonate in blood was slightly elevated (24.1--25.7--25.0 mmol/l; p= 0.008) during the MH treatment period, when compared to pre- and post-treatment control periods. It is suggested that MH treatment reduced the complications associated with indwelling catheters due to reduction in urine pH, bacteriuria, and pyuria. Blockage of catheters is thought to be due to intraluminal salt precipitations with trapping of clumps, and is primarily not correlated with urine viscosity.

Aged↗

Randomized double-blind study of prophylactic methenamine hippurate treatment of patients with indwelling catheters.

Catheterized patients with heavy bacteriuria and short catheter life received a short-course of antimicrobial therapy and then were allocated at random to double-blind treatment for 5 months with methenamine hippurate (MH), 1 g three times daily, or placebo 1 g 3 times daily. MH treatment reduced the number of catheter blockages and instances of spontaneous removal of the catheter, and doubled catheter life (p less than 0.001). It is suggested that the doubled catheter life was due to the reduced bacteriuria and lowering of urinary pH.

Aged↗

Use of methenamine hippurate in male infertility.

In sperm-freezing for infertility patients with low sperm counts, we found that semen containing high numbers of white blood cells, greater than 15 to 20 per high-power field, was both poor in quality and difficult to freeze. In examining these patients carefully, we found that many had smoldering prostatitis and some an overt history of prostatitis. We treated them with methenamine hippurate using various methods and found that by decreasing the white blood cell count in their semen, we could frequently improve the semen quality, particularly sperm motility, also achieve pregnancies in some cases.

Cell Count↗

Methenamine silver staining quantitative digital histochemistry in chronic allograft nephropathy.

Renal function and final outcome of renal allografts have been correlated with irreversible damage. This study describes a quantitative histochemical method relying on periodic acid methenamine silver (PAMS) staining of all renal compartments. Among 60 renal allograft biopsies from 43 patients, 15 biopsies showing pure chronic allograft nephropathy were selected to determine PAMS-stained area percentage (SAP), using image analysis with quantitative histochemistry. Of the 15 cases, 9 (60%) were grade I and 6 (40%) were grade II chronic allograft nephropathy (CAN). The mean serum creatinine (sCr) value was 1.86 +/- 0.47 for allograft biopsies. The mean (+/-SD) SAP for the implantation biopsies was 10.58 +/- 1.87%, and for allograft biopsies 25.26 +/- 9.67 (P <<.000). The serum creatinine (sCr) values for grade I versus II CAN were 1.63 +/- 0.24 versus 2.20 +/- 0.54 mg/dL, respectively (P=.019), and SAP values were 18.97 +/- 0.24 versus 34.7 +/- 5.89 (P=.003). There was a strong positive correlation between sCr values and SAP (P=.005; r=0.64). These findings show the PAMS approach to be a useful alternative method for reflecting damage in more than one compartment of the renal tissue. Also, the method can discriminated implantation and allograft biopsies as well as grade I and II CAN cases. The series is small for a multivariate analysis of the value of SAP measurements in PAMS-stained sections as a prognosticator, but the data support its use.

Biopsy↗

Quantitative digital histochemistry with methenamine silver staining in renal allograft biopsies excluding pure chronic allograft nephropathy cases.

Deterioration of renal function is correlated with irreversible damage in chronic diseases. Recently we described a digital quantitative histochemistry method, relying on periodic acid methenamine silver (PAMS) staining to determine the chronic renal lesions. This index was strongly correlated with progressive deterioration of renal function in grafts with chronic allograft nephropathy (CAN). Herein the method has been applied to a cohort of renal allografts which were biopsied for various reasons, we sought to highlight its value to quantify chronic graft damage. Forty-four renal allograft biopsies from 37 patients with elevated serum creatinine values (SCr) underwent light microscopic image analysis (Mediscope, Dokuz Eylül University, Clinical Engineering Department, Izmir, Turkey) of the PAMS-stained area percentage (SAP). SCr was recorded at four intervals to overcome acute effects: the under SCr value before (SCr1) and after a biopsy within 3 months (SCr3), SCr at the time of the biopsy (SCr2), and the latest value (SCr4). The PAMS-SAP scores were strongly associated with increased interstitial fibrosis and tubular atrophy Banff scores (Kruskal-Wallis test, P = .006 and P = .003, respectively). There was a moderate positive correlation between PAMS and SCr3 (Pearson correlation test, P = .04, r = .312), and a strong positive correlation between time from transplantation to biopsy (Pearson correlation test, P << .000, r = .532). The present results show that PAMS-SAP seems to be of value to quantify renal scarring in allograft biopsies, reflecting four compartments. The strong correlation with time is noteworthy especially as a probable reflection of aging of the renal allograft.

Anti-Infective Agents, Urinary↗

Long-term prophylaxis of urinary infections in women: comparative trial of trimethoprim, methenamine hippurate and topical povidone-iodine.

We randomized 64 patients with a history of recurrent urinary tract infections among 3 regimens of long-term (1 year) prophylactic treatment: 20 were given 100 mg. trimethoprim at night, 25 received 1,000 mg. methenamine hippurate every 12 hours and 19 were asked to cleanse the perineum (especially the periurethral area) twice daily with povidone-iodine solution. The progress of patients in terms of urinary symptoms and/or bacteriuria, changes in periurethral flora, side effects, and hematological and biochemical profiles was followed at regular intervals. All treatments were effective in reducing the incidence of symptomatic attacks when compared to the 12 months immediately before therapy and there was little to choose between the individual regimens on this account. However, trimethoprim was tolerated better than were the other 2 treatment regimens. In the group given trimethoprim most of the breakthrough infections (71.4 per cent) that occurred were caused by trimethoprim-resistant organisms (usually Escherichia coli), while in the other 2 groups the incidence of trimethoprim-resistant organisms causing infection was low (2.7 per cent). Treatment with trimethoprim reduced significantly the periurethral colonization of Escherichia coli.

Administration, Topical↗