Fundamental equations and hypotheses governing glomerular hemodynamics.

The glomerular filtration rate (GFR) is the outcome of glomerular hemodynamics, influenced by a series of parameters: renal plasma flow, resistances of afferent arterioles and efferent arterioles (EAs), hydrostatic pressures in the glomerular capillary and Bowman's capsule, and plasma colloid osmotic pressure in the glomerular capillary. Although mathematical models have been proposed to predict the GFR at both the single-nephron level and the two-kidney system level using these parameters, mathematical equations governing glomerular filtration have not been well-established because of two major problems. First, the two-kidney system-level models are simply extended from the equations at the single-nephron level, which is inappropriate in epistemology and methodology. Second, the role of EAs in maintaining the normal GFR is underappreciated. In this article, these two problems are concretely elaborated, which collectively shows the need for a shift in epistemology toward a more holistic and evolving way of thinking, as reflected in the concept of the complex adaptive system (CAS). Then, we illustrate eight fundamental mathematical equations and four hypotheses governing glomerular hemodynamics at both the single-nephron and two-kidney levels as the theoretical foundation of glomerular hemodynamics. This illustration takes two steps. The first step is to modify the existing equations in the literature and establish a new equation within the conventional paradigm of epistemology. The second step is to formulate four hypotheses through logical reasoning from the perspective of the CAS (beyond the conventional paradigm). Finally, we apply the new equation and hypotheses to comprehensively analyze glomerular hemodynamics under different conditions and predict the GFR. By doing so, some concrete issues are eliminated. Unresolved issues are discussed from the perspective of the CAS and a desinger's view. In summary, this article advances the theoretical study of glomerular dynamics by 1) clarifying the necessity of shifting to the CAS paradigm; 2) adding new knowledge/insights into the significant role of EAs in maintaining the normal GFR; 3) bridging the significant gap between research findings and physiology education; and 4) establishing a new and advanced foundation for physiology education.

HIGH THROUGHPUT QUANTITATION OF HUMAN NEUTROPHIL RECRUITMENT AND FUNCTIONAL RESPONSES IN AN AIR-BLOOD BARRIER ARRAY.

Dysregulated neutrophil recruitment drives many pulmonary diseases, but most preclinical screening methods are unsuited to evaluate pulmonary neutrophilia, limiting progress towards therapeutics. Namely, high throughput therapeutic screening systems typically exclude critical neutrophilic pathophysiology, including blood-to-lung recruitment, dysfunctional activation, and resulting impacts on the air-blood barrier. To meet the conflicting demands of physiological complexity and high throughput, we developed an assay of 96-well Leukocyte recruitment in an Air-Blood Barrier Array (L-ABBA-96) that enables in vivo -like neutrophil recruitment compatible with downstream phenotyping by automated flow cytometry. We modeled acute respiratory distress syndrome (ARDS) with neutrophil recruitment to 20 ng/mL epithelial-side interleukin 8 (IL-8) and found a dose dependent reduction in recruitment with physiologic doses of baricitinib, a JAK1/2 inhibitor recently FDA-approved for severe COVID-19 ARDS. Additionally, neutrophil recruitment to patient-derived cystic fibrosis sputum supernatant induced disease-mimetic recruitment and activation of healthy donor neutrophils and upregulated endothelial e-selectin. Compared to 24-well assays, the L-ABBA-96 reduces required patient sample volumes by 25 times per well and quadruples throughput per plate. Compared to microfluidic assays, the L-ABBA-96 recruits two orders of magnitude more neutrophils per well, enabling downstream flow cytometry and other standard biochemical assays. This novel pairing of high-throughput in vitro modeling of organ-level lung function with parallel high-throughput leukocyte phenotyping substantially advances opportunities for pathophysiological studies, personalized medicine, and drug testing applications.

Politically related stress and low-birth-weight infants among Arab, Asian, Hispanic, non-Hispanic Black, and non-Hispanic White women in Michigan.

BACKGROUND: Despite the high cost of low birth weight and the persistent challenge of racial inequities affecting the Arab American community, there has been limited research to identify and examine risk factors for these inequities with validated data on Arab American ethnicity and recent population stressors. OBJECTIVES: This study examined whether the 2016 presidential election is associated with low birth weight among non-Hispanic White, Arab American, Hispanic, and non-Hispanic Black women. DESIGN: This population-based study of singleton births in Michigan (2008-2017) used an algorithm to identify mothers who were of Arab descent. METHODS: We used logistic regression to estimate odds ratios and 95% confidence intervals for the association between race/ethnicity and the odds of low birth weight. We examined whether these associations differed before and after the 2016 presidential election and according to maternal education. RESULTS: There were 1,019,738 births, including 66,272 (6.5%) classified as low birth weight. The odds of having a low-birth-weight infant were higher among all minority women compared to non-Hispanic White women. The association was similar before and after the 2016 presidential election and stronger among women with higher levels of education. CONCLUSION: This is the first study to estimate low birth weight among Arab American women in the context of political events. There are opportunities for future studies to discuss this issue in depth.

Topographical and zonal patterns of T2 relaxation in osteoarthritic tibial cartilage by low- and high-resolution MRI.

PURPOSE: This study aimed to establish the topographical and zonal T2 patterns of multi-resolution MRI in medial tibial cartilage in a canine model of osteoarthritis (OA), initiated by the anterior cruciate ligament (ACL) transection surgery, and studied after 8-weeks and 12-weeks post-surgery. METHODS: Articular cartilage from healthy, two stages of contralateral, and of OA knees were quantitatively imaged by the MRI T2 protocols at two imaging resolutions (100 and 17.6 µm/pixel). The zonal T2 changes at five topographical locations (anterior (AMT), exterior (EMT), posterior (PMT), central (CMT) and interior (IMT) medial tibia) and subsequent two averaged regions (covered by meniscus and exposed) were analyzed. At each location, full-thickness cartilage was studied in four sub-tissue zones (superficial, transitional, upper and lower radial zones). RESULTS: Tissue degradation can be detected by measurable changes of T2, which is resolution- and orientation-dependent. T2 changes ranging from +28.82% increase (SZ, PMT) to -23.15% decrease (RZ1, AMT) in healthy to disease (8C), with the largest increase of T2 in the surface tissue. Various location-dependent patterns of degradation are found over the tibial surface, most commonly shown in early-stage OA (8C) on the anterior site, different from the posterior. Finally, the contralateral cartilage has specific degradation patterns, different from those in OA cartilage. CONCLUSIONS: This is the first quantitative and highest multi-resolution characterization of cartilage at five topographical locations over the medial tibial plateau with fine zonal resolution in an animal model of OA, which would benefit future investigation of human OA in clinics.

Untangling the relatedness among correlations, part III: Inter-subject correlation analysis through Bayesian multilevel modeling for naturalistic scanning.

While inter-subject correlation (ISC) analysis is a powerful tool for naturalistic scanning data, drawing appropriate statistical inferences is difficult due to the daunting task of accounting for the intricate relatedness in data structure as well as handling the multiple testing issue. Although the linear mixed-effects (LME) modeling approach (Chen et al., 2017a) is capable of capturing the relatedness in the data and incorporating explanatory variables, there are a few challenging issues: 1) it is difficult to assign accurate degrees of freedom for each testing statistic, 2) multiple testing correction is potentially over-penalizing due to model inefficiency, and 3) thresholding necessitates arbitrary dichotomous decisions. Here we propose a Bayesian multilevel (BML) framework for ISC data analysis that integrates all regions of interest into one model. By loosely constraining the regions through a weakly informative prior, BML dissolves multiplicity through conservatively pooling the effect of each region toward the center and improves collective fitting and overall model performance. In addition to potentially achieving a higher inference efficiency, BML improves spatial specificity and easily allows the investigator to adopt a philosophy of full results reporting. A dataset of naturalistic scanning is utilized to illustrate the modeling approach with 268 parcels and to showcase the modeling capability, flexibility and advantages in results reporting. The associated program will be available as part of the AFNI suite for general use.

Progress in the Use of Induced Pluripotent Stem Cell-Derived Neural Cells for Traumatic Spinal Cord Injuries in Animal Populations: Meta-Analysis and Review.

Induced pluripotent stem cells (iPSCs) are cells genetically reprogrammed from somatic cells, which can be differentiated into neurological lineages with the aim to replace or assist damaged neurons in the treatment of spinal cord injuries (SCIs) caused by physical trauma. Here, we review studies addressing the functional use of iPSC-derived neural cells in SCIs and perform a meta-analysis to determine if significant motor improvement is restored after treatment with iPSC-derived neural cells compared with treatments using embryonic stem cell (ESC)-derived counterpart cells and control treatments. Overall, based on locomotion scales in rodents and monkeys, our meta-analysis indicates a therapeutic benefit for SCI treatment using neural cells derived from either iPSCs or ESCs, being this of importance due to existing ethical and immunological complications using ESCs. Results from these studies are evidence of the successes and limitations of iPSC-derived neural cells in the recovery of motor capacity. Stem Cells Translational Medicine 2019;8:681&693.

A Meta-Analysis Detailing Overall Sexual Function and Orgasmic Function in Women Undergoing Midurethral Sling Surgery for Stress Incontinence.

INTRODUCTION: More than 200,000 midurethral slings are placed yearly for stress urinary incontinence. Approximately 14% to 20% of women experience worsening sexual function overall after these procedures. We postulated that sling placement injures neural pathways regulating "female prostate" tissue within the anterior vaginal wall. AIM: To perform a meta-analysis for overall sexual function and orgasm using validated questionnaires for transvaginal tape, transobturator tape, and their variants. METHODS: Effect sizes of preoperative and postoperative questionnaire scores for overall sexual function and orgasm were calculated. Random-effects models were selected for meta-analyses. Statistical analysis involved determination of the ratio of total heterogeneity to total variability. MAIN OUTCOME MEASURES: Differences of overall sexual and orgasm functions were calculated by subtracting postoperative sling scores from preoperative sling scores. Forest plots of effect sizes were performed. RESULTS: Sixty-seven percent of midurethral sling procedures analyzed showed no change or improvement in overall sexual function postoperatively, whereas 33% of studies analyzed for orgasm function showed improvement after the procedure. For transvaginal tape, mean total sexual function and orgasm postoperative scores were significantly higher than preoperative scores. For transobturator tape, mean total postoperative score was significantly higher than the preoperative score; however, the mean orgasm postoperative score was not significantly higher than the preoperative score, possibly because of variability in transobturator tape data. CONCLUSION: There is a discrepancy between postoperative sexual satisfaction and orgasmic function after midurethral sling surgery. Although overall sexual function remained the same or improved for most women, orgasmic function in only one third of cases improved overall, with most women experiencing no change or deterioration in orgasmic function. Dissection for, and placement of, the midurethral sling can compromise the neural integrity of the anterior vaginal wall, thereby detrimentally affecting the periurethral prostate tissue that is essential to the orgasmic response. We propose that this surgical procedure can compromise orgasmic function in some women. Szell N, Komisaruk B, Goldstein SW, et al. A Meta-Analysis Detailing Overall Sexual Function and Orgasmic Function in Women Undergoing Midurethral Sling Surgery for Stress Incontinence. Sex Med 2017;5:e84-e93.

Elevated CXC chemokines in urine noninvasively discriminate OAB from UTI.

Overlapping symptoms of overactive bladder (OAB) and urinary tract infection (UTI) often complicate the diagnosis and contribute to overprescription of antibiotics. Inflammatory response is a shared characteristic of both UTI and OAB and here we hypothesized that molecular differences in inflammatory response seen in urine can help discriminate OAB from UTI. Subjects in the age range of (20-88 yr) of either sex were recruited for this urine analysis study. Urine specimens were available from 62 UTI patients with positive dipstick test before antibiotic treatment. Six of these patients also provided urine after completion of antibiotic treatment. Subjects in cohorts of OAB (n = 59) and asymptomatic controls (n = 26) were negative for dipstick test. Urinary chemokines were measured by MILLIPLEX MAP Human Cytokine/Chemokine Immunoassay and their association with UTI and OAB was determined by univariate and multivariate statistics. Significant elevation of CXCL-1, CXCL-8 (IL-8), and CXCL-10 together with reduced levels for a receptor antagonist of IL-1A (sIL-1RA) were seen in UTI relative to OAB and asymptomatic controls. Elevated CXCL-1 urine levels predicted UTI with odds ratio of 1.018 and showed a specificity of 80.77% and sensitivity of 59.68%. Postantibiotic treatment, reduction was seen in all CXC chemokines with a significant reduction for CXCL-10. Strong association of CXCL-1 and CXCL-10 for UTI over OAB indicates mechanistic differences in signaling pathways driving inflammation secondary of infection in UTI compared with a lack of infection in OAB. Urinary chemokines highlight molecular differences in the paracrine signaling driving the overlapping symptoms of UTI and OAB.

Topographical variations in zonal properties of canine tibial articular cartilage due to early osteoarthritis: a study using 7-T magnetic resonance imaging at microscopic resolution.

OBJECTIVE: Our aim was to determine topographical variations in zonal properties of articular cartilage over the medial tibia in an experimental osteoarthritis (OA) model using 7-T magnetic resonance imaging (MRI). MATERIALS AND METHODS: An anterior cruciate ligament (ACL)-transection canine model was subjected to study at 8 (six) and 12 (seven) weeks after the surgery. Each medial tibia was divided into five topographical locations. For each specimen, T2 relaxation (at 0° and 55°) was quantified at microscopic resolution. The imaging data grouped the five locations into two topographical areas (meniscus-covered and -uncovered). RESULTS: The T2 (55°) bulk values from the meniscus-covered area were significantly lower than those from the uncovered area. The total cartilage thicknesses on the meniscus-covered area were significantly thinner than those on the meniscus-uncovered area. Significant differences in the T2 (0°) values were observed in most thicknesses of the four subtissue zones and whole-tissue from the uncovered area, while the same significant changes were detected in the superficial zone from the meniscus-covered area. CONCLUSION: By quantifying high-resolution imaging data both topographically and depth-dependently (zonal-wise), this study demonstrates that the rate of disease progression varies topographically over the medial tibia. Future correlation with OA pathology could lead to better detection of early OA.

Loading-induced changes on topographical distributions of the zonal properties of osteoarthritic tibial cartilage--A study by magnetic resonance imaging at microscopic resolution.

The topographical distributions of the zonal properties of articular cartilage over the medial tibia from an experimental osteoarthritis (OA) model were evaluated as a function of external loading by microscopic Magnetic Resonance Imaging (µMRI). T2 relaxation times and cartilage thicknesses were measured at 17.6 µm resolution from 118 specimens, which came from thirteen dogs (six 8-week and seven 12-week after surgery), with and without mechanical loading. In addition, bulk mechanical modulus was measured topographically from each tibia surface. The total thickness decreased significantly under the external loading, in which the relative thickness of the superficial zone (SZ) and the transitional zone (TZ) increased whereas the radial zones (RZs) decreased. In the bulk data, T2(55°) decreased significantly (p<0.001) at all OA-time-points, but T2(0°) decreased without significance (p>0.05) at 8-week. Complex relationships were found in the zonal tissue properties as a function of external loading with the progress of OA. T2 in the superficial zone changed more profoundly than the same properties in the radial zone as a function of external loading at all OA time-points. This study confirms that OA affects the load-induced changes in the molecular distribution and structure of cartilage, which are both depth-dependent and topographically distributed. Such detailed knowledge of mechanobiological changes in specific tibial cartilage zones and locations with OA progress could improve the early detection of the subtle softening of cartilage that accompanies pre-clinical stages of OA.

Topographical variations of the strain-dependent zonal properties of tibial articular cartilage by microscopic MRI.

The topographical variations of the zonal properties of canine articular cartilage over the medial tibia were evaluated as the function of external loading by microscopic magnetic resonance imaging (µMRI). T2 and T1 relaxation maps and GAG (glycosaminoglycan) images from a total of 70 specimens were obtained with and without the mechanical loading at 17.6 µm depth resolution. In addition, mechanical modulus and water content were measured from the tissue. For the bulk without loading, the means of T2 at magic angle (43.6 ± 8.1 ms), absolute thickness (907.6 ± 187.9 µm) and water content (63.3 ± 9.3%) on the meniscus-covered area were significantly lower than the means of T2 at magic angle (51.1 ± 8.5 ms), absolute thickness (1251.6 ± 218.4 µm) and water content (73.2 ± 5.6%) on the meniscus-uncovered area. However GAG (86.0 ± 15.3 mg/ml) on the covered area was significantly higher than GAG (70.0 ± 8.8 mg/ml) on the uncovered area. Complex relationships were found in the tissue properties as the function of external loading. The tissue parameters in the superficial zone changed more profoundly than the same properties in the radial zone. The tissue parameters in the meniscus-covered areas changed differently when comparing with the same parameters in the uncovered areas. This project confirms that the load-induced changes in the molecular distribution and structure of cartilage are both depth-dependent and topographically distributed. Such detailed knowledge of the tibial layer could improve the early detection of the subtle softening of the cartilage that will eventually lead to the clinical diseases such as osteoarthritis.

Association of inflammaging (inflammation + aging) with higher prevalence of OAB in elderly population.

INTRODUCTION: Although epidemiology studies consistently report increased prevalence of overactive bladder (OAB) with age, an accurate deciphering of causative links between the two entities remains elusive. Studies on aged rodent bladder have so far yielded contradictory results on age-associated changes in muscarinic receptors, which highlight the challenge posed by species differences in understanding OAB pathology. We hypothesized that age-related biochemical changes in bladder leading to altered bladder function will be reflected in altered urinary proteome of elderly OAB patients. METHODS: Single time point urine specimens were obtained from 140 OAB patients in the age range of 25-90 years of either sex coming routinely to the urology clinics. Eight chemokines in urine were measured by MILLIPLEX MAP human cytokine/chemokine multiplex immunoassay and ELISA. Multivariate and univariate statistical analyses were done to determine association of age with urinary chemokines in OAB patients. RESULTS: In agreement with age-dependent higher prevalence of OAB, the logistic regression of the data also revealed the significant association of OAB symptoms with age [odds ratio (OR) 1.12; 95 % CI, (1.072, 1.187), p = 0.0001]. Univariate analysis of 8 urinary proteins revealed an age-associated elevation of NGF (nerve growth factor) in 137 out of 140 OAB patients [Pearson r = 0.274; 95 %CI (0.112-0.422); p = 0.001]. Modest correlation with age was also noted for MCP-1 (monocyte chemoattractant protein-1), which was detected in 115 OAB patients, and the remaining chemokines were undetectable in nearly two-third of OAB patients included in our cohort. CONCLUSIONS: Based on our findings, we postulate that age-associated biochemical changes may accentuate the inflammation associated with OAB. Urinary NGF elevation in elderly OAB patients may be a homeostatic response to counter the senescence of bladder nerves and arrest the progression of OAB into detrusor hyperactivity with impaired contractility. Likewise, elevation of MCP-1 may be related to decreased muscle mass and increased content of adipose tissue in bladder of elderly OAB patients. Urinary NGF and MCP-1 can serve as surrogate markers for monitoring age-associated biochemical changes and the effect of therapeutic interventions in OAB patients.

Core strength training using a combination of home exercises and a dynamic sling system for the management of low back pain in pre-professional ballet dancers: a case series.

Estimates of low back pain prevalence in USA ballet dancers range from 8% to 23%. Lumbar stabilization and extensor muscle training has been shown to act as a hypoalgesic for low back pain. Timing and coordination of multifidi and transverse abdominis muscles are recognized as important factors for spinal stabilization. The purpose of this study was to explore the effects of training methods using home exercises and a dynamic sling system on core strength, disability, and low back pain in pre-professional ballet dancers. Five participants were randomly assigned to start a traditional unsupervised lumbar stabilization home exercise program (HEP) or supervised dynamic sling training to strengthen the core and lower extremities. Measurements were taken at baseline and at weeks 3 and 6 for disability using the patient specific functional scale (PSFS), pain using the Numerical Pain Rating System (NPRS), core strength and endurance using timed plank, side-plank, and bridge positions, and sciatic nerve irritability using the straight leg raise (SLR). Data were analyzed using descriptive statistics. From initial to final measurements, all participants demonstrated an improvement in strength and SLR range, and those with initial pain and disability reported relief of symptoms. These results suggest that dynamic sling training and a HEP may help to increase strength, decrease pain, and improve function in dancers without aggravating sciatic nerve irritation.

Personal health behaviors and role-modeling attitudes of physical therapists and physical therapist students: a cross-sectional study.

BACKGROUND: Physical therapists have been encouraged to engage in health promotion practice. Health professionals who engage in healthy behaviors themselves are more apt to recommend those behaviors, and patients are more motivated to change their behaviors when their health care provider is a credible role model. OBJECTIVE: The purpose of this study was to describe the health behaviors and role-modeling attitudes of physical therapists and physical therapist students. DESIGN: This study was a descriptive cross-sectional survey. METHODS: A national sample of 405 physical therapists and 329 physical therapist students participated in the survey. Participants' attitudes toward role modeling and behaviors related to physical activity, fruit and vegetable consumption, abstention from smoking, and maintenance of a healthy weight were measured. Wilcoxon rank sum tests were used to examine differences in attitudes and behaviors between physical therapists and physical therapist students. RESULTS: A majority of the participants reported that they engage in regular physical activity (80.8%), eat fruits and vegetables (60.3%), do not smoke (99.4%), and maintain a healthy weight (78.7%). Although there were no differences in behaviors, physical therapist students were more likely to believe that role modeling is a powerful teaching tool, physical therapist professionals should "practice what they preach," physical activity is a desirable behavior, and physical therapist professionals should be role models for nonsmoking and maintaining a healthy weight. LIMITATIONS: Limitations of this study include the potential for response bias and social desirability bias. CONCLUSIONS: Physical therapists and physical therapist students engage in health-promoting behaviors at similarly high rates but differ in role-modeling attitudes.

384 hanging drop arrays give excellent Z-factors and allow versatile formation of co-culture spheroids.

We previously reported the development of a simple, user-friendly, and versatile 384 hanging drop array plate for 3D spheroid culture and the importance of utilizing 3D cellular models in anti-cancer drug sensitivity testing. The 384 hanging drop array plate allows for high-throughput capabilities and offers significant improvements over existing 3D spheroid culture methods. To allow for practical 3D cell-based high-throughput screening and enable broader use of the plate, we characterize the robustness of the 384 hanging drop array plate in terms of assay performance and demonstrate the versatility of the plate. We find that the 384 hanging drop array plate performance is robust in fluorescence- and colorimetric-based assays through Z-factor calculations. Finally, we demonstrate different plate capabilities and applications, including: spheroid transfer and retrieval for Janus spheroid formation, sequential addition of cells for concentric layer patterning of different cell types, and culture of a wide variety of cell types.

The effect of tibio-femoral traction mobilization on passive knee flexion motion impairment and pain: a case series.

The purpose of this case series was to explore the effects of tibio-femoral (TF) manual traction on pain and passive range of motion (PROM) in individuals with unilateral motion impairment and pain in knee flexion. Thirteen participants volunteered for the study. All participants received 6 minutes of TF traction mobilization applied at end-range passive knee flexion. PROM measurements were taken before the intervention and after 2, 4, and 6 minutes of TF joint traction. Pain was measured using a visual analog scale with the TF joint at rest, at end-range passive knee flexion, during the application of joint traction, and immediately post-treatment. There were significant differences in PROM after 2 and 4 minutes of traction, with no significance noted after 4 minutes. A significant change in knee flexion of 25.9°, which exceeded the MDC(95,) was found when comparing PROM measurements pre- to final intervention. While pain did not change significantly over time, pain levels did change significantly during each treatment session. Pain significantly increased when the participant's knee was passively flexed to end range; it was reduced, although not significantly, during traction mobilization; and it significantly decreased following traction. This case series supports TF joint traction as a means of stretching shortened articular and periarticular tissues without increasing reported levels of pain during or after treatment. In addition, this is the first study documenting the temporal aspects of treatment effectiveness in motion restoration.

Lack of association of the VEGF gene promoter (-634 G-->C and -460 C-->T) polymorphism and the risk of advanced retinopathy of prematurity.

BACKGROUND: Recently, it has been reported that genetic polymorphism (-634 G-->C and -460 C-->T) in the promoter region of the vascular endothelial growth factor (VEGF) gene can influence the progression of retinopathy of prematurity (ROP). In order to evaluate its general applicability as a screening procedure in clinics and to replicate the above result, we have undertaken the following study. METHODS: We have analyzed a cohort of 61 patients with advanced ROP (stage 4 and 5) along with 61 normal controls for the VEGF gene promoter polymorphism. For this purpose, blood samples were collected from each patient and leukocyte DNA was isolated. Genomic DNA was amplified by the polymerase chain reaction (PCR) method with two pairs of primers designed to amplify separately the promoter region (containing -634 G-->C and -460 C-->T polymorphism) of the VEGF gene. The amplified product was subjected to restriction enzyme digestion. The base change in the restriction site was further confirmed by a BigDye terminator cycle sequencing of the amplified product. RESULTS: Our analysis suggests that there is no significant difference in allelic frequency of the VEGF gene between normal subjects and patients with advanced ROP in our cohort. CONCLUSION: Our results do not support the association of the VEGF gene promoter polymorphism and the risk of advanced ROP. In order to adapt this method for the identification of high-risk infants in clinics in the future, a large-scale study involving a mixed ethnically diverse population is much needed.

Statistical analysis of differences in the Raman spectra of polymorphs.

Raman spectroscopy is a useful tool for identifying polymorphs of pharmaceutical compounds. One limitation of the technique is that the small differences in Raman spectra require confirmation of polymorphs by other methods. Fourteen compounds, both commercial and proprietary pharmaceutical compounds and their polymorphs, were analyzed by Raman microscopy. By using descriptive statistics and analysis of variance (ANOVA), several methods are proposed that provide an approach for comparing the spectra of suspected polymorphs. Because it is difficult to determine the exact amount that a peak may shift before two forms should be considered different; a guideline of a shift greater than 1.6/cm(-1) is proposed. A standard ANOVA analysis is used to compare individual peaks both within and between polymorphs, as well as an alternative method that proposes the use of a total ANOVA table that considers the entire spectrum. Both methods have their advantages and disadvantages, but they provide a starting point for the comparison of a large number of spectra, and effectively differentiate between polymorphs of a given compound. The method accurately identified true polymorphs in all cases, but showed a bias towards misidentifying some samples as polymorphs when they were in fact the same form. This bias was not significant and even in these situations, the magnitude of the calculated F values was a useful indicator of whether the result was a false positive or not.

Evaluation of the vibratory feeder method for assessment of powder flow properties.

The flow properties of pharmaceutical powders and blends used in solid oral dosage forms are an important consideration during dosage form development. The vibratory feeder method, a flow measurement technique that quantifies avalanche flow, has been adapted for measurement of the flow properties of common pharmaceutical powders used in solid oral dosage forms. The flow properties of 17 different powders were measured with the instrument, and the results are reported as a powder flow index (PFI). The PFI trends of the powders correlate well with flow properties reported in the literature. The flow properties of the powders were also measured with a commercially available avalanche instrument, the Aero-Flow, and the results were reported as the mean time to avalanche (MTA). Since the two instruments analyze the avalanche by different algorithms, the results were compared with nonparametric statistical evaluation of ranked data, and they were found to be in excellent agreement. A recommended procedure for measurement of powder flow with the vibratory feeder is presented.