Andre J. Sommer

Randall’s plaque of patients with nephrolithiasis begins in basement membranes of thin loops of Henle

Our purpose here is to test the hypothesis that Randalls plaques, calcium phosphate deposits in kidneys of patients with calcium renal stones, arise in unique anatomical regions of the kidney, their formation conditioned by specific stone-forming pathophysiologies. To test this hypothesis, we performed intraoperative biopsies of plaques in kidneys of idiopathic-calcium-stone formers and patients with stones due to obesity-related bypass procedures and obtained papillary specimens from non-stone formers after nephrectomy. Plaque originates in the basement membranes of the thin loops of Henle and spreads from there through the interstitium to beneath the urothelium. Patients who have undergone bypass surgery do not produce such plaque but instead form intratubular hydroxyapatite crystals in collecting ducts. Non-stone formers also do not form plaque. Plaque is specific to certain kinds of stone-forming patients and is initiated specifically in thin-limb basement membranes by mechanisms that remain to be elucidated.

Mechanism of Formation of Human Calcium Oxalate Renal Stones on Randall's Plaque

Although calcium oxalate (CaOx) renal stones are known to grow attached to renal papillae, and specifically to regions of papillae that contain Randalls plaque (interstitial apatite deposits), the mechanisms of stone overgrowth on plaque are not known. To investigate the problem, we have obtained biopsy specimens from two stone patients that included an attached stone along with its tissue base and have studied the ultrastructural features of the attachment point using light and transmission electron microscopy, Fourier transform infrared spectroscopy (μ‐FTIR), and immunohistochemical analysis. The epithelium is disrupted at the attachment site. The denuded plaque that borders on the urinary space attracts an envelope of ribbon‐like laminates of crystal and organic matrix arising from urine ions and molecules. Into the matrix of this ribbon grow amorphous apatite crystals that merge with and give way to the usual small apatite crystals imbedded in stone matrix; eventually CaOx crystals admix with apatite and become the predominant solid phase. Over time, urine calcium and oxalate ions gradually overgrow on the large crystals forming the attached stone. Anat Rec, 290:1315‐1323, 2007.

Attenuated Total Internal Reflection Infrared Mapping Microspectroscopy Using an Imaging Microscope

Attenuated total internal reflection (ATR) infrared mapping microspectroscopy using an infrared microscope and a focal plane array is investigated and reported. The study demonstrates the advantages of conducting ATR microspectroscopy using a focal plane array detector. These benefits include the rapid acquisition of molecular specific images, ease of sample preparation, and increased spatial resolution. An experimental determination of the spatial resolution found that the combined system operates very close to the diffraction limit, and a 4 magnification factor associated with the germanium internal reflection element was realized. Experiments conducted on several polymer samples and a biological sample demonstrate the future viability of the method.

Discrimination among Carbonate Minerals by Raman Spectroscopy Using the Laser Microprobe

Raman spectroscopy has been used to distinguish the individual carbonate minerals belonging to the calcite, dolomite, and aragonite structural groups. With the use of the in situ laser microprobe technique, it is demonstrated that particle-size effects do not hinder the mineral identification and that high precision in line position is achieved. Spectra can be obtained from samples in any form and provide rapid, nondestructive analyses on a microscopic scale of the hard-to-discriminate carbonate minerals.

Nondestructive analysis of urinary calculi using micro computed tomography.

BackgroundMicro computed tomography (micro CT) has been shown to provide exceptionally high quality imaging of the fine structural detail within urinary calculi. We tested the idea that micro CT might also be used to identify the mineral composition of urinary stones non-destructively.MethodsMicro CT x-ray attenuation values were measured for mineral that was positively identified by infrared microspectroscopy (FT-IR). To do this, human urinary stones were sectioned with a diamond wire saw. The cut surface was explored by FT-IR and regions of pure mineral were evaluated by micro CT to correlate x-ray attenuation values with mineral content. Additionally, intact stones were imaged with micro CT to visualize internal morphology and map the distribution of specific mineral components in 3-D.ResultsMicro CT images taken just beneath the cut surface of urinary stones showed excellent resolution of structural detail that could be correlated with structure visible in the optical image mode of FT-IR. Regions of pure mineral were not difficult to find by FT-IR for most stones and such regions could be localized on micro CT images of the cut surface. This was not true, however, for two brushite stones tested; in these, brushite was closely intermixed with calcium oxalate. Micro CT x-ray attenuation values were collected for six minerals that could be found in regions that appeared to be pure, including uric acid (3515 – 4995 micro CT attenuation units, AU), struvite (7242 – 7969 AU), cystine (8619 – 9921 AU), calcium oxalate dihydrate (13815 – 15797 AU), calcium oxalate monohydrate (16297 – 18449 AU), and hydroxyapatite (21144 – 23121 AU). These AU values did not overlap. Analysis of intact stones showed excellent resolution of structural detail and could discriminate multiple mineral types within heterogeneous stones.ConclusionsMicro CT gives excellent structural detail of urinary stones, and these results demonstrate the feasibility of identifying and localizing most of the common mineral types found in urinary calculi using laboratory CT.

Histopathology and surgical anatomy of patients with primary hyperparathyroidism and calcium phosphate stones

Using a combination of intra-operative digital photography and micro-biopsy we measured renal cortical and papillary changes in five patients with primary hyperparathyroidism and abundant calcium phosphate kidney stones. Major tissue changes were variable papillary flattening and retraction, dilation of the ducts of Bellini, and plugging with apatite deposits of the inner medullary collecting ducts and ducts of Bellini. Some of the papillae in two of the patients contained plentiful large interstitial deposits of Randalls plaque and where the deposits were most plentiful we found overgrowth of the attached stones. Hence, this disease combines features previously described in brushite stone formers--dilation, plugging of ducts and papillary deformity--with the interstitial plaque and stone overgrowth characteristic of routine idiopathic calcium oxalate stone formers, suggesting that these two patterns can coexist in a single patient.

Microbial effects in promoting the smectite to illite reaction: Role of organic matter intercalated in the interlayer

Abstract Cysteine and toluene as model organic molecules were intercalated into Fe-rich smectite (nontronite, NAu-2). The illitization of these intercalated smectites as induced by microbial reduction of structural Fe3+ was investigated. Iron-reducing bacterium Shewanella putrefaciens CN32 was incubated with lactate as the sole electron donor and structural Fe3+ in cysteine- and toluene-intercalated NAu-2 (referred to as cysteine-NAu-2 and toluene-NAu-2 hereafter) as the sole electron acceptor. Anthraquinone- 2, 6-disulfonate (AQDS) was used as an electron shuttle in bicarbonate buffer. The extent of Fe3+ reduction in cysteine-NAu-2 and toluene-NAu-2 was 15.7 and 5.4%, respectively, compared to 20.5% in NAu-2 without organic matter intercalation. In the bioreduced NAu-2, X-ray diffraction, and scanning and transmission electron microscopy did not detect any discrete illite, although illite/ smectite mixed layer or high charge smectite phases were observed. In bioreduced cysteine-NAu-2, discrete illite and siderite formed. In contrast, bioreduction of toluene-NAu-2 did not result in any mineralogical changes. The contrasting bioreduction results between cysteine- and toluene-intercalated nontronite may be ascribed to the nature of organic matter-bacteria interactions. Whereas cysteine is an essential amino acid for bacteria and can also serve as an electron shuttle, thus enhancing the extent of Fe3+ bioreduction and illitization, toluene is toxic and inhibits Fe3+-reducing activity. This study, therefore, highlights the significant role of organic matter in promoting the smectite to illite reaction under conditions typical of natural environments (i.e., non-growth condition for bacteria).

Renal histopathology and crystal deposits in patients with small bowel resection and calcium oxalate stone disease

We present here the anatomy and histopathology of kidneys from 11 patients with renal stones following small bowel resection, including 10 with Crohns disease and 1 resection in infancy for unknown cause. They presented predominantly with calcium oxalate stones. Risks of formation included hyperoxaluria (urine oxalate excretion greater than 45 mg per day) in half of the cases, and acidic urine of reduced volume. As was found with ileostomy and obesity bypass, inner medullary collecting ducts (IMCDs) contained crystal deposits associated with cell injury, interstitial inflammation, and papillary deformity. Cortical changes included modest glomerular sclerosis, tubular atrophy, and interstitial fibrosis. Randalls plaque (interstitial papillary apatite) was abundant, with calcium oxalate stone overgrowth similar to that seen in ileostomy, idiopathic calcium oxalate stone formers, and primary hyperparathyroidism. Abundant plaque was compatible with the low urine volume and pH. The IMCD deposits all contained apatite, with calcium oxalate present in three cases, similar to findings in patients with obesity bypass but not an ileostomy. The mechanisms for calcium oxalate stone formation in IMCDs include elevated urine and presumably tubule fluid calcium oxalate supersaturation, but a low calcium to oxalate ratio. However, the mechanisms for the presence of IMCD apatite remain unknown.

The advantages of an attenuated total internal reflection infrared microspectroscopic imaging approach for kidney biopsy analysis

The benefits of an attenuated total reflection Fourier transform infrared (ATR-FTIR) imaging approach for kidney biopsy analysis are described. Biopsy sections collected from kidney-stone formers are analyzed at the initial stages of stone development to provide insights into stone growth and formation. The majority of tissue analysis currently conducted with IR microspectroscopy is performed with a transflection method. The research presented in this manuscript demonstrates that ATR overcomes many of the disadvantages of transflection or transmission measurements for tissue analysis including an elimination of spectral artifacts. When kidney biopsies with small mineral inclusions are analyzed with a transflection approach, specular reflection and the Christiansen effect (anomalous dispersion) can occur, leading to spectral artifacts. Another effect specific to the analysis of mineral inclusions present in kidney biopsies is known as the reststrahlen effect whereby the inclusions become strong reflectors near an absorption band. ATR eliminates these effects by immersing the sample in a high index medium. Additionally, the focused beam size for ATR is decreased by a factor of four when a germanium internal reflection element is used, allowing the acquisition of spectra from small mineral inclusions several micrometers in diameter. If quantitative analysis of small mineral inclusions is ultimately desired, ATR provides the photometrically accurate spectra necessary for quantification.

Diffraction-Induced Stray Light in Infrared Microspectroscopy and Its Effect on Spatial Resolution

Model experiments were conducted in an effort to quantitatively assess the extent of stray light, resulting from diffraction, in an FT-IR microscope system. The effects of stray light were studied under conditions employing different aperturing modes, aperture sizes, and wavelengths of light. Results and consequences of the findings are discussed with respect to the spatial resolution and quantitative integrity of the data obtainable in mapping analyses of multilayer polymer laminates.