An integrative correlation of myopathology, phenotype and genotype in late onset Pompe disease.

AIMS: Pompe disease is caused by pathogenic mutations in the alpha 1,4-glucosidase (GAA) gene and in patients with late onset Pome disease (LOPD), genotype-phenotype correlations are unpredictable. Skeletal muscle pathology includes glycogen accumulation and altered autophagy of various degrees. A correlation of the muscle morphology with clinical features and the genetic background in GAA may contribute to the understanding of the phenotypic variability. METHODS: Muscle biopsies taken before enzyme replacement therapy were analysed from 53 patients with LOPD. On resin sections, glycogen accumulation, fibrosis, autophagic vacuoles and the degree of muscle damage (morphology-score) were analysed and the results were compared with clinical findings. Additional autophagy markers microtubule-associated protein 1A/1B-light chain 3, p62 and Bcl2-associated athanogene 3 were analysed on cryosections from 22 LOPD biopsies. RESULTS: The myopathology showed a high variability with, in most patients, a moderate glycogen accumulation and a low morphology-score. High morphology-scores were associated with increased fibrosis and autophagy highlighting the role of autophagy in severe stages of skeletal muscle damage. The morphology-score did not correlate with the patient's age at biopsy, disease duration, nor with the residual GAA enzyme activity or creatine-kinase levels. In 37 patients with LOPD, genetic analysis identified the most frequent mutation, c.-32-13T>G, in 95%, most commonly in combination with c.525delT (19%). No significant correlation was found between the different GAA genotypes and muscle morphology type. CONCLUSIONS: Muscle morphology in LOPD patients shows a high variability with, in most cases, moderate pathology. Increased pathology is associated with more fibrosis and autophagy.

The critical size of a defect in the glenoid causing anterior instability of the shoulder after a Bankart repair, under physiological joint loading.

AIMS: Patients with recurrent anterior dislocation of the shoulder commonly have an anterior osseous defect of the glenoid. Once the defect reaches a critical size, stability may be restored by bone grafting. The critical size of this defect under non-physiological loading conditions has previously been identified as 20% of the length of the glenoid. As the stability of the shoulder is load-dependent, with higher joint forces leading to a loss of stability, the aim of this study was to determine the critical size of an osseous defect that leads to further anterior instability of the shoulder under physiological loading despite a Bankart repair. PATIENTS AND METHODS: Two finite element (FE) models were used to determine the risk of dislocation of the shoulder during 30 activities of daily living (ADLs) for the intact glenoid and after creating anterior osseous defects of increasing magnitudes. A Bankart repair was simulated for each size of defect, and the shoulder was tested under loading conditions that replicate in vivo forces during these ADLs. The critical size of a defect was defined as the smallest osseous defect that leads to dislocation. RESULTS: The FE models showed a high risk of dislocation during ADLs after a Bankart repair for anterior defects corresponding to 16% of the length of the glenoid. CONCLUSION: This computational study suggests that bone grafting should be undertaken for an anterior osseous defect in the glenoid of more than 16% of its length rather than a solely soft-tissue procedure, in order to optimize stability by restoring the concavity of the glenoid.

Predictive value of ex vivo biodynamic imaging in determining response to chemotherapy in dogs with spontaneous non-Hodgkin's lymphomas: a preliminary study.

Biodynamic imaging (BDI) is a novel phenotypic cancer profiling technology which optically characterizes changes in subcellular motion within living tumor tissue samples in response to ex vivo treatment with cancer chemotherapy drugs. The purpose of this preliminary study was to assess the ability of ex vivo BDI to predict in vivo clinical response to chemotherapy in ten dogs with naturally-occurring non-Hodgkin's lymphomas. Pre-treatment tumor biopsy samples were obtained from all dogs and treated ex vivo with doxorubicin (10 µM). BDI measured six dynamic biomarkers of subcellular motion from all biopsy samples at baseline and at regular intervals for 9 h following drug application. All dogs subsequently received doxorubicin to treat their lymphomas. Best overall response to and progression-free survival time following chemotherapy were recorded for all dogs. Receiver operating characteristic (ROC) curves were used to determine accuracy and identify possible cut-off values for the BDI-measured biomarkers which could accurately predict those dogs' cancers that would and would not respond to doxorubicin chemotherapy. One biomarker (designated 'MEM') showed 100% discriminative capability for predicting clinical response to doxorubicin (area under the ROC curve = 1.00, 95% CI 0.692-1.000), while other biomarkers also showed promising predictive capability. These preliminary findings suggest that ex vivo BDI can accurately predict treatment outcome following doxorubicin chemotherapy in a spontaneous animal cancer model, and is worthy of further investigation as a technology for personalized cancer medicine.

Point-of-Care Clinical Ultrasound for Medical Students.

PURPOSE: Our institution has recently implemented a point-of-care (POC) ultrasound training program, consisting of an e-learning course and systematic practical hands-on training. The aim of this prospective study was to evaluate the learning outcome of this curriculum. MATERIALS AND METHODS: 16 medical students with no previous ultrasound experience comprised the study group. The program covered a combination of 4 well-described point-of-care (POC) ultrasound protocols (focus assessed transthoracic echocardiography, focused assessment with sonography in trauma, lung ultrasound, and dynamic needle tip positioning for ultrasound-guided vascular access) and it consisted of an e-learning course followed by 4 h of practical hands-on training. Practical skills and image quality were tested 3 times during the study: at baseline, after e-learning, and after hands-on training. RESULTS: Practical skills improved for all 4 protocols; after e-learning as well as after hands-on training. The number of students who were able to perform at least one interpretable image of the heart increased from 7 at baseline to 12 after e-learning, p<0.01, and to all 16 students after hands-on-training, p<0.01. The number of students able to cannulate an artificial vessel increased from 3 to 8 after e-learning and to 15 after hands-on training. CONCLUSION: Medical students with no previous ultrasound experience demonstrated a considerable improvement in practical skill after interactive e-learning and 4 h of hands-on training.

White matter damage is related to ataxia severity in SCA3.

Spinocerebellar ataxia type 3 (SCA3) is the most frequent inherited cerebellar ataxia in Europe, the US and Japan, leading to disability and death through motor complications. Although the affected protein ataxin-3 is found ubiquitously in the brain, grey matter atrophy is predominant in the cerebellum and the brainstem. White matter pathology is generally less severe and thought to occur in the brainstem, spinal cord, and cerebellar white matter. Here, we investigated both grey and white matter pathology in a group of 12 SCA3 patients and matched controls. We used voxel-based morphometry for analysis of tissue loss, and tract-based spatial statistics (TBSS) on diffusion magnetic resonance imaging to investigate microstructural pathology. We analysed correlations between microstructural properties of the brain and ataxia severity, as measured by the Scale for the Assessment and Rating of Ataxia (SARA) score. SCA3 patients exhibited significant loss of both grey and white matter in the cerebellar hemispheres, brainstem including pons and in lateral thalamus. On between-group analysis, TBSS detected widespread microstructural white matter pathology in the cerebellum, brainstem, and bilaterally in thalamus and the cerebral hemispheres. Furthermore, fractional anisotropy in a white matter network comprising frontal, thalamic, brainstem and left cerebellar white matter strongly and negatively correlated with SARA ataxia scores. Tractography identified the thalamic white matter thus implicated as belonging to ventrolateral thalamus. Disruption of white matter integrity in patients suffering from SCA3 is more widespread than previously thought. Moreover, our data provide evidence that microstructural white matter changes in SCA3 are strongly related to the clinical severity of ataxia symptoms.

Spinocerebellar ataxia type 17 associated with an expansion of 42 glutamine residues in TATA-box binding protein gene.

BACKGROUND: Spinocerebellar ataxia type 17 (SCA17) is caused by abnormal expansions of CAG/CAA trinucleotides within the TATA-box binding protein gene (TBP). The currently accepted critical threshold of abnormal expansions is ≥43. OBJECTIVE: To investigate the minimal CAG/CAA expansion within the TBP in SCA17. RESULTS: 285 patients with autosomal-dominant ataxia were examined, and abnormal or borderline expansions of CAG/CAA within TBP in eight cases were found. Of those, four patients from three families had exactly 42 CAG/CAA trinucleotides, that is, one codon less than the currently accepted critical threshold of 43. The four patients presented with a relatively benign phenotype. All had dysdiadochokinesia and dysarthria. Mild gait ataxia was observed in three of the four patients. CONCLUSION: The reference definition of at least 43 CAG/CAA codons for pathological SCA17 alleles should be lowered to 42.

Three-dimensional holographic imaging of living tissue using a highly sensitive photorefractive polymer device.

Photorefractive materials are dynamic holographic storage media that are highly sensitive to coherent light fields and relatively insensitive to a uniform light background. This can be exploited to effectively separate ballistic light from multiply-scattered light when imaging through turbid media. We developed a highly sensitive photorefractive polymer composite and incorporated it into a holographic optical coherence imaging system. This approach combines the advantages of coherence-domain imaging with the benefits of holography to form a high-speed wide-field imaging technique. By using coherence-gated holography, image-bearing ballistic light can be captured in real-time without computed tomography. We analyzed the implications of Fourier-domain and image-domain holography on the field of view and image resolution for a transmission recording geometry, and demonstrate holographic depth-resolved imaging of tumor spheroids with 12 microm axial and 10 microm lateral resolution, achieving a data acquisition speed of 8 x 10(5) voxels/s.

Combined fluorescent and interferometric detection of protein on a BioCD.

We perform simultaneous interferometric and fluorescent detection of molecular protein layers on a BioCD. The 488 nm excitation wavelength of fluorescein also provides the interferometric detection channel that operates in a common-path in-line configuration in the condition of phase quadrature set by a thermal oxide on silicon. The simultaneous acquisition of both channels enables a direct correlation between bound mass and fluorescent surface density, which we compare in forward- and reverse-phase immunoassays. Scaling mass sensitivities for immunoassays measured in the interferometric and fluorescent channels are 15 pg/mm and 1.5 pg/mm, respectively, when applied to gel-printed periodic antibody patterns detected in the frequency domain from the spinning disc. These sensitivities are limited by the inhomogeneities of the print. While fluorescence is subject to bleaching, the interferometry signal is robust under long-term laser illumination.

Common-path interferometric detection of protein monolayer on the BioCD.

The bio-optical compact disk (BioCD) is an optical biosensor that performs common-path molecular interferometry of patterned proteins on a disk spinning at high speed. The common-path configuration makes it ultrastable and allows surface height precision below 10 pm. In this paper we show that two complementary interferometric quadrature conditions exist simultaneously that convert the modulus and phase of the reflection coefficient, modulated by protein patterns on the disk surface, into intensity modulation at the detector. In the far field they separate into spatially symmetric and antisymmetric intensity modulation in response to the local distribution of protein. The antisymmetric response is equivalent to differential phase-contrast detection, and the symmetric response is equivalent to in-line (IL) common-path interferometry. We measure the relative sensitivities of these orthogonal channels to printed protein patterns on disk structures that include thermal oxide on silicon and Bragg dielectric stacks. The scaling mass sensitivity of the IL channel on oxide on silicon was measured to be 0.17 pg/mm.

Stages of sleep pathology in spinocerebellar ataxia type 2 (SCA2).

BACKGROUND: Autosomal dominant spinocerebellar ataxia type 2 (SCA2) bears clinical and neuropathologic similarities to sporadic multisystem atrophy (MSA) or Parkinson disease, in which sleep pathology is well documented. However, those clinical entities have a marked variability of the reported sleep disturbances, and their etiology is heterogeneous. In contrast, the study of SCA2 provides an opportunity to examine a molecularly homogeneous patient group, in which disease stages can be defined not only based on disease duration and ataxia scores, but also with regard to modulatory effects of mutation size. OBJECTIVE: To examine the presence and progression of sleep pathology in SCA2. METHODS: We analyzed eight patients with disease durations of 3 to 31 years, all with medium size SCA2 expansions (CAG 38 to 49), using clinical scores, sleep interviews, and video-polysomnography (VPSG) recordings. RESULTS: Almost all patients reported good subjective sleep quality and negated incidents of REM behavior disorder (RBD). At early disease stages, however, REM without atonia in four patients' VPSG suggested subclinical RBD. This was accompanied by a consistent reduction of REM density. In three patients at later SCA2 stages, REM sleep was undetectable, whereas slow wave sleep (SWS) was markedly increased at the cost of light sleep. Periodic leg movements, apnea, or hypopnea were not prominent. CONCLUSIONS: Progressive loss of dream recall in spinocerebellar ataxia type 2 was found and correlated with stages of REM more than non-REM pathology in video-polysomnography. These stages correspond to the progressive atrophy from the pons, nigrostriatal projection, and locus ceruleus to the thalamus.

Reversible ischaemia after raising a radial forearm flap with ulceration of three fingers in a cigarette smoker.

The radial forearm flap is a standard method for the reconstruction of intraoral defects of soft tissues. We report the case of a middle-aged man who developed ischaemia in three fingers after a fasciocutaneous radial flap had been raised. The preoperative Allen test to diagnose occlusion of radial or ulnar artery was satisfactory. Soon after the operation the patient resumed smoking and four weeks later he developed ulcers on the thumb, index, and middle fingers. Only after he had stopped smoking and been given acetylsalicylic acid and heparin did blood flow and capillary hemoglobin oxygenation increase. As a result, his radial fingers recovered completely.

An autosomal dominant ataxia maps to 19q13: Allelic heterogeneity of SCA13 or novel locus?

The autosomal dominant spinocerebellar ataxias (ADCAs) represent a growing and heterogeneous disease phenotype. Clinical characterization of a three-generation Filipino family segregating a dominant ataxia revealed cerebellar signs and symptoms. After elimination of known spinocerebellar ataxia (SCA) loci, a genome-wide linkage scan revealed a disease locus in a 4-cM region of 19q13, with a 3.89 lod score. This region overlaps and reduces the SCA13 locus. However, this ADCA is clinically distinguishable from SCA13.

[Simultaneous noninvasive monitoring for radial forearm and fibula flaps using laser Doppler flowmetry and tissue spectrophotometry]. / Simultanes nichtinvasives Monitoring mit Laser-Doppler-Flussmessung und Gewebespektrometrie bei fasziokutanen Radialislappen und osteokutanen Fibulatransplantaten.

AIM: In the literature currently available monitoring devices are usually divided into two major groups: those for monitoring perfusion and those for measuring tissue oxygenation. The O(2)C (oxygen to see) system combines these two ways of monitoring free flap viability. The aim of this prospective study was to determine the necessity of flap revision and when unnecessary revision can be avoided. Another point of interest was the question of whether critical values for the successful course of free flaps could be defined and in addition whether such values would differ for different flap types. PATIENTS AND METHODS: In a prospective study 82 free flaps (61 radial forearm flaps and 21 fibula flaps) were monitored with the O(2)C monitoring unit. Measurements were carried out intraoperatively and postoperatively up to 14 days. RESULTS: Perfusion compromise occurred in 12 (14.6%) of 82 monitored free flaps. Operative exploration was performed in seven cases, in five of them successfully. Five flaps (three radial forearm and two fibula flaps) were lost due to vascular compromise, which led to an overall success rate of 93.4%. Venous congestion was identified by a rapid increase in hemoglobin concentration of more than 30%. An abrupt decline of blood flow and hemoglobin oxygenation indicated arterial occlusion. Vascular complications were detected in all cases prior to clinical assessment with no false positive or negative results. For radial forearm flaps a hemoglobin oxygenation of 15%, a superficial flow of 10 AU, and a deep flow of 20 AU were identified as minimum values for flap viability. For fibula flaps a hemoglobin oxygenation of 10%, a superficial flow of 5 AU, and a deep flow of 15 AU were determined as minimum values. CONCLUSION: O(2)C combines laser Doppler flowmetry and tissue spectrophotometry and for the first time allows simultaneous measurement of the microcirculatory parameters including blood flow, flow velocity, hemoglobin concentration, and hemoglobin oxygenation. We found this new noninvasive technique to be a reliable and accurate method for evaluating flap viability and improving the success rate in free flap transfer.

[Limitations of blood supply to the skin flap in face lift surgery]. / Limitationen der Blutversorgung des Hautlappens beim Facelifting Eine anatomische Studie.

BACKGROUND: Besides nerve injuries or hematomas, one of the main risks of face lift surgery is the development of skin necrosis. We therefore investigated which arteries contribute to the blood supply of the face lift skin flap and how extensively mobilization can be performed before arterial cutaneous circulation is endangered. MATERIAL AND METHOD: In six cadavers, selective dye injections into the branches of the carotid external artery were performed to determine the location of their vascular territories in relation to the face lift region. In another 12 cadavers, face lift flaps were raised with different extension of the undermined skin area before dye injections were performed. RESULTS: It was shown that the facial artery provides blood supply to the area of the face lift flap by perforating vessels in the paranasal, perioral and submental regions and the transverse facial artery by a perforating vessel below the malar bone prominence. Although the perforating vessel of the transverse facial artery was transected by standard mobilization, the face lift flaps were sufficiently perfused via the paranasal and perioral perforating vessels of the facial artery alone. CONCLUSION: We conclude that undermining of the face lift flap should not exceed the nasolabial fold to preserve the perforating vessels of the facial artery in the centrofacial region.

Attenuation of leukocyte sequestration by selective blockade of PECAM-1 or VCAM-1 in murine endotoxemia.

BACKGROUND: Molecular mechanisms regulating leukocyte sequestration into the tissue during endotoxemia and/or sepsis are still poorly understood. This in vivo study investigates the biological role of murine PECAM-1 and VCAM-1 for leukocyte sequestration into the lung, liver and striated skin muscle. METHODS: Male BALB/c mice were injected intravenously with murine PECAM-1 IgG chimera or monoclonal antibody (mAb) to VCAM-1 (3 mg/kg body weight); controls received equivalent doses of IgG2a (n = 6 per group). Fifteen minutes thereafter, 2 mg/kg body weight of Salmonella abortus equi endotoxin was injected intravenously. At 24 h after the endotoxin challenge, lungs, livers and striated muscle of skin were analyzed for their myeloperoxidase activity. To monitor intravital leukocyte-endothelial cell interactions, fluorescence videomicroscopy was performed in the skin fold chamber model of the BALB/c mouse at 3, 8 and 24 h after injection of endotoxin. RESULTS: Myeloperoxidase activity at 24 h after the endotoxin challenge in lungs (12,171 +/- 2,357 mU/g tissue), livers (2,204 +/- 238 mU/g) and striated muscle of the skin (1,161 +/- 110 mU/g) was significantly reduced in both treatment groups as compared to controls, with strongest attenuation in the PECAM-1 IgG treatment group. Arteriolar leukocyte sticking at 3 h after endotoxin (230 +/- 46 cells x mm(-2)) was significantly reduced in both treatment groups. Leukocyte sticking in postcapillary venules at 8 h after endotoxin (343 +/- 69 cells/mm2) was found reduced only in the VCAM-1-mAb-treated animals (215 +/- 53 cells/mm2), while it was enhanced in animals treated with PECAM-1 IgG (572 +/- 126 cells/mm2). CONCLUSION: These data show that both PECAM-1 and VCAM-1 are involved in endotoxin-induced leukocyte sequestration in the lung, liver and muscle, presumably through interference with arteriolar and/or venular leukocyte sticking.

[Botulinum toxin for treatment of gustatory sweating. A prospective randomized study]. / Botulinumtoxin zur Behandlung des gustatorischen Schwitzens. Eine prospektive randomisierte Therapiestudie.

BACKGROUND: Botulinum toxin A has meanwhile become a proven method for treatment of gustatory sweating (focal hyperhidrosis, Frey's syndrome). Clear-cut recommendations regarding dosage of botulinum toxin A in Frey's syndrome are currently not available. The aim of this prospective randomized study therefore was to investigate botulinum toxin A with respect to its efficacy in Frey's syndrome, the ideal dose yielding maximal duration of the effect, and patient contentedness as well as unwanted side effects in patients of the Clinic of OMF Surgery at the Ruhr-University of Bochum. PATIENTS AND METHOD: Twenty patients suffering from severe Frey's syndrome as a result of operations of the parotid gland were examined with the starch iodine test according to Minor. The gustatory skin areas were re-examined after intracutaneous injection of botulinum toxin A for up to 1 year. The patients ( n=20) were randomly assigned to two different treatment groups (group I: 2 MU/cm(2), n=10; group II: 3 MU/cm(2), n=10). RESULTS: Mean sweating skin areas in the two treatment groups ranged between 39+/-9 and 32+/-12 cm(2), respectively. A single injection of 3 MU botulinum toxin A resulted in a nearly complete blockade of gustatory sweating for the observation period of 1 year. In the group treated with 2 MU botulinum toxin A, 44% of the total gustatory skin areas were still sweating, thus necessitating a second injection of botulinum toxin A in these patients. CONCLUSION: Intracutaneous injection of botulinum toxin A represents a highly effective and minimally invasive procedure for the treatment of Frey's syndrome. This study shows for the first time that a dosage of 3 MU/cm(2) of botulinum toxin A achieves a complete and reliable blockade of gustatory sweating lasting for at least 12 months. This dose may therefore be recommended for treatment of this syndrome.

[Importance of microcirculation in plastic surgery]. / Die Bedeutung der Mikrozirkulation in der Plastischen Chirurgie.

The article summarizes distinct microcirculatory models for use in surgical research with a special interest to plastic surgery. Methods for the quantitative analysis of the microcirculation in burns, flaps and wounds are presented. Vascularization of biomaterials can be observed by means of the dorsal skinfold chamber model in hamsters and mice. Developing capillary sprouts can be assessed by means of videomicroscopy and angiogenetic drugs can be tested using this model. Hairless mice allow for direct, long-term observation of the microcirculation in burns as well as during healing of dermal wounds. The pathophysiology of diabetic wound healing can also be studied. A mouse model to assess flap microcirculation during ischemia/reperfusion injury with special emphasis on platelet/endothelium interaction in vivo is described. Platelets adherent to the inner vessel wall are known to trigger compromised perfusion in flaps. The model allows us to test anti-thrombotic drugs. The use of a special microscopic device (OPS imaging) allows us to study the microcirculation at sites of burn injury and chronic wounds in humans. Microcirculatory research in plastic surgery has increased the understanding of the pathophysiology of vascularization of biomaterials, wound healing and ischemia/reperfusion.

[Management of zygomatic fractures via a transconjunctival approach with lateral canthotomy while preserving the lateral ligament]. / Versorgung von Jochbeinfrakturen über einen transkonjunktivalen Zugang mit lateraler Kanthotomie unter Schonung des lateralen Lidbandes.

INTRODUCTION: This study reports on a modified approach to treat zygomatic fracture. For the surgical approach, a transconjunctival incision and, instead of a lateral canthotomy, a superficial incision of the skin and subcutaneous tissue was used, preserving the lateral ligament. The results are compared with those of previous techniques, especially the subciliary incision. PATIENTS: In a prospective study, 30 patients (9 females and 21 males, mean age 32.1 years) with zygomatic fractures were operated using this approach. Mean follow-up time was 6 months after removal of the plates. Fractures were caused by trauma due to fighting, bicycle falls, or sport accidents. Follow-up radiographs were used to evaluate the position of the zygoma after reduction and rigid fixation. RESULTS: Reduction and rigid fixation of the fractures were possible to perform in all cases. The access to the orbital floor and the exposure of the inferior and lateral rim were satisfactory and an additional latero-orbital cut was not necessary. The disadvantage of the complete incision of the lateral ligament with the necessity of intraoperative refixation and possible ectropium of the eyelid could be avoided. Injuries or infections of the cornea or the bulbus did not occur. Postoperative complications such as scar formation resulting in entropium or ectropium were not seen. CONCLUSION: These results show that the transconjunctival approach with lateral superficial incision preserving the lateral ligament for treatment of zygomatic fractures is satisfactory in all cases of rigid fixation of both inferior and lateral rims. Because of its esthetic and functional advantages this approach has become standard in our department.

Spinning-disk self-referencing interferometry of antigen-antibody recognition.

A gold ridge microstructure fabricated to a height of lambda/8 on a high-reflectivity substrate behaves as a wave-front-splitting self-referencing interferometer in phase quadrature when illuminated by a Gaussian laser beam and observed in the far field along the optic axis. When immuno-gammaglobulin (IgG) antibodies are selectively immobilized on the gold microstructure, they recognize and bind to a specific antigen, which shifts the relative optical phase of the interferometer and modifies the far-field diffracted intensity. We detect bound antigen interferometrically on spinning disks at a sampling rate of 100 kHz and verify the interferometric nature of the signal by using two quadratures of opposite sign to rule out effects of dynamic light scattering. Strong molecular recognition is demonstrated by the absence of binding to nontarget molecules but strong signal change in response to a specific antigen. This BioCD has the potential to be applied as a spinning-disk interferometric immunoassay and biosensor.

Time-dependent speckle in holographic optical coherence imaging and the health of tumor tissue.

Holographic optical coherence imaging acquires en face images from successive depths inside scattering tissue. In a study of multicellular tumor spheroids the holographic features recorded from a fixed depth are observed to be time dependent, and they may be classified as variable or persistent. The ratio of variable to persistent features, as well as speckle correlation times, provides quantitative measures of the health of the tissue. Studies of rat osteogenic sarcoma tumor spheroids that have been subjected to metabolic and cross-polymerizing poisons provide quantitative differentiation among healthy, necrotic, and poisoned tissue. Organelle motility in healthy tissue appears as super-Brownian laser speckle, whereas chemically fixed tissue exhibits static speckle.