Rapid aneuploidy screening with fluorescence in-situ hybridisation: is it a sufficiently robust stand-alone test for prenatal diagnosis?

OBJECTIVES: To assess the clinical utility of fluorescence in-situ hybridisation with chromosomes 13, 18, 21, X and Y as a stand-alone test in detecting chromosomal abnormalities, and the types of chromosomal abnormalities missed. DESIGN: Retrospective analysis. SETTING: A restructured Government hospital in Singapore and an academic hospital in the United States. PARTICIPANTS: Cytogenetic data of prenatal specimens and results of fluorescence in-situ hybridisation of 5883 patients performed between January 2000 and August 2007 were reviewed. RESULTS: Fluorescence in-situ hybridisation detected 558 (9.5%) patients with chromosomal abnormalities. Abnormal ultrasounds (70%) and maternal serum screens (21%) were the most indicative of chromosomal abnormalities. When comparing fluorescence in-situ hybridisation data with karyotype results for the five chromosomes of interest, the sensitivity and specificity were 99.3% and 99.9%, respectively. When comparing fluorescence in-situ hybridisation data with karyotype results for all chromosomes, the sensitivity decreased to 86.8%, whereas the specificity remained at 99.9%. Of 643 cases with karyotype abnormalities, 85 were fluorescence in-situ hybridisation-negative (false negative rate, 13.2%), which included structural rearrangements, chromosome mosaicism, and other trisomies. Despite abnormal ultrasound indications, fluorescence in-situ hybridisation missed 32 cases which included structural rearrangements, mosaicisms, and other trisomies. CONCLUSION: This study does not support fluorescence in-situ hybridisation as a stand-alone test. Institutions supporting fluorescence in-situ hybridisation as a stand-alone test must seriously consider the risks of a missed diagnosis.

[Psychotherapeutic care of dysphonic patients]. / Psychotherapeutische Weiterbetreuung stimmgestörter Patienten.

The diagnosis of vocal dysfunction is a prerequisite for referring a patient for specialized medical therapy. This mostly requires a holistic approach, and should also consider related symptoms and the social environment of the patient. Regulations on psychotherapy, promulgated in 1999, have provided clarity in relation to the indications for treatment in an inpatient and outpatient basis. Within the German health care system, every individual can now directly contact an officially certified psychotherapist (so called psychological psychotherapist) for probationary sessions, aimed at determining a basis and indication for further psychotherapy. In case of psychotherapy provided from others than physicians, an additional medical report from a physician is obligatory. Patients with functional and psychogenic voice problems often believe that organic disorders are the reason for their dysphonia. Thus, when caring for voice disordered patients and sending them to a psychotherapist, it is of great importance that physicians facilitate their referral by reacting in an empathic, fear reducing, and professional way.

Data analyst technician: an innovative role for the pharmacy technician.

The development of an innovative role for the pharmacy technician is described. The role of the pharmacy technician was based on a needs assessment and the expertise of the pharmacy technician selected. Initial responsibilities of the technician included chart reviews, benchmarking surveys, monthly financial impact analysis, initiative assessment, and quality improvement reporting. As the drug-use and disease-state management (DUDSM) program expanded, pharmacist activities increased, requiring the expansion of data analyst technician (DAT) duties. These new responsibilities included participation in patient assessment, data collection and interpretation, and formulary enforcement. Most recently, technicians' expanded duties include maintenance of a physician compliance profiling database, quality improvement reporting and graphing, active role in patient risk assessment and database management for adult vaccination, and support of financial impact monitoring for other institutions within the health system. This pharmacist-technician collaboration resulted a threefold increase in patient assessments completed per day. In addition, as the DUDSM program continues to expand across the health system, an increase in DAT resources from 0.5 to 1.0 full-time equivalent was obtained. The role of the DAT has increased the efficiency of the DUDSM program and has provided an innovative role for the pharmacy technician.

Dehydroepiandrosterone, dehydroepiandrosterone-sulfate, and cortisol concentrations in intensive care unit patients.

STUDY OBJECTIVE: This purpose of this study was to determine whether severity of illness, as defined by the intensive care unit (ICU) admission APACHE II (updated Acute Physiology and Chronic Health Evaluation) score, is correlated with early morning cortisol, dehydroepiandrosterone (DHEA), and/or dehydroepiandrosterone-sulfate (DHEA-S) concentrations. DESIGN: Early morning concentrations of DHEA, DHEA-S, and cortisol were determined within 24 hrs of admission and compared with admission APACHE II scores. SETTING: Medical (MICU), neurologic (NICU), and surgical (SICU) intensive care units of the University of Pittsburgh Medical Center. PATIENTS: A total of 191 men and women ranging in age from 16 to 93 yrs. All had been admitted to an ICU. MEASUREMENTS AND MAIN RESULTS: Statistically significant correlations between APACHE II scores and cortisol were observed for women in the MICU and SICU (r = .68, p = .0001; r = .35 p = .017, respectively) and for men in the NICU (r = .55, p = .003) and the SICU (r = .29, p = .036). The correlations between APACHE II scores and DHEA concentration data were statistically significant for women in the MICU (r = .37, p = .047) and SICU (r = .43, p = .002), as was the correlation between APACHE II and DHEA-S concentrations among women in the SICU (r = .38, p = .008). Although not statistically significant, a similar relationship was observed in the smaller group of NICU women (r = .40, p = .099). Each correlation was essentially unchanged when adjusted for age. CONCLUSION: These data show a positive correlation between APACHE II and cortisol concentrations in all groups except the MICU men. Also evident is the positive correlation between APACHE II scores and DHEA and DHEA-S concentrations in women, but not in men.

A quantitative output-cost ratio in voice production.

A quantitative output-cost ratio (OCR) is proposed for objective use in voice production and is defined as the ratio of the acoustic output intensity to the collision intensity of the vocal folds. Measurement of the OCR is demonstrated in a laboratory experiment using 5 excised larynges and a transducer designed for use on human subjects. Data were gathered at constant fundamental frequency (150 Hz). Subglottal pressure was varied from 1.0 to 1.6 kPa, and glottal width at the vocal processes was varied from a pressed condition to a 2-mm gap. The OCR was plotted as a function of glottal width. With no vocal tract, the excised larynx experiments yielded a broad maxima in the OCR curves, across all subglottal pressure conditions, at about 0.6 mm. Computer simulations indicate that sharper maxima may occur when the influence of the vocal tract is taken into account. The potential clinical utility of the OCR is discussed for treatment of a wide range of voice disorders, including those involving both hyper- and hypoadduction.

Vocal fold impact stress analysis.

Vocal fold impact stress (force/area) has been implicated as a factor possibly contributing to the formation of nodules and polyps. The force of impact of a moving body is related to its acceleration. Since the mass of the folds is relatively constant, one expects impact force to be directly proportional to acceleration. A measure that reflects the relative displacement of the vocal folds is photoglottography (PGG). The velocity and acceleration of the folds are easily obtained by calculating the first and second derivatives of the PGG displacement waveform. This study, therefore, compared the second derivative of the PGG signal with simultaneously measured impact stress in an excised canine larynx model. Glottal transillumination (PGG) was measured with a subglottic transducer. A miniature force transducer placed in the midline between the vocal folds measured impact stress at the midglottal position. For nine different larynges, there was a positive and linear relationship between the second derivative of PGG and impact stress. The statistically significant results support the hypothesis that the second derivative of PGG m ay provide a use fulnoninvasive way to estimate relative vocal fold impact stress.

Strobophotoglottographic transillumination as a method for the analysis of vocal fold vibration patterns.

The purpose of this exploratory study was to determine if laryngeal transillumination in combination with stroboscopy (strobophotoglottography; SPGG) is useful for (1) the visualization of vocal fold vibration (VFV) opening patterns, (2) the localization of initial vocal fold opening in horizontal glottal thirds (anterior, midmembranous, and posterior), (3) determination of the temporal correspondence of the so-called electroglottography (EGG)-knee and initial vocal fold separation, and, finally, (4) automatized quantitative measurements of glottal area function within endoscopic images. With stroboscopic transillumination, initial inferior vocal fold separation was detectable during the "closed" phase, where the vocal folds were still closed in the upper portion and therefore initial inferior vocal fold separation could not be visualized with usual laryngoscopy techniques. In the horizontal plane within similar fundamental frequencies in modal voice registers in two male subjects, localization of initial glottal opening depended on the voice types used (soft, normal, or pressed phonation). We found zipperlike posterior-to-anterior openings, initial midmembranous openings, initial anterior openings, as well as simultaneous initial opening of all three portions in the two healthy male adults examined. This technique proved to add temporal and spatial information to vocal fold opening patterns and extends our examination techniques to the very beginning of vocal fold opening at the inferior portion. Simultaneous electroglottogram tracking and comparison with bidirectionally illuminated stroboscopic images revealed a time-locked correspondence of the EGG-knee with the aforementioned initial inferior vocal fold separation. Bidirectional illumination combined with digital color extraction techniques allowed for image separation of subglottally and supraglottally illuminated structures. This facilitated vocal fold contour detection and automatized image processing, for example, for determination of glottal area function, and is considered to be a further step to objective automatized quantitative measurements within endoscopic images.

Implementation of a drug-use and disease-state management program.

A drug-use and disease-state management (DUDSM) program was instituted in 1996 at a teaching hospital associated with a large nonprofit health care system. The program's goals are to optimize pharmacotherapeutic regimens, evaluate health outcomes of identified disease states, and evaluate the economic impact of pharmacotherapeutic options for given disease states by developing practice guidelines. Through a re-engineering process, resources within the pharmacy department were identified that could be devoted to the DUDSM program, including the use of clinical pharmacy specialists, promotion of staff pharmacists into the DUDSM program, a pharmacy technician, and information systems support. A strength of the program is its systematic approach for developing and implementing new initiatives, as well as monitoring compliance with all initiatives on an ongoing basis. The initiative-design process incorporates continuous quality improvement principles, outcome design and evaluation, competency assessment for all pharmacists, multidisciplinary collaboration, and sophisticated information systems. Seventy-five initiatives have been implemented, ranging from simple dose-optimization strategies for specific drugs to complicated practice guidelines for managing specific disease states. Improved patient outcomes have been documented, including reduced length of stay, postsurgical wound infection, adverse drug reactions, and medication errors. Documented cost savings exceeded $4 million annually for fiscal years 1996-97 through 1999-2000. Overall compliance with DUDSM initiatives exceeds 80%, and physician service profiling has been initiated to monitor variant prescribing. The DUDSM program has successfully integrated practice guidelines into therapeutic decision-making, resulting in improved patient-care outcomes and cost savings.

Qualitative and quantitative analysis of voice onset by means of a multidimensional voice analysis system (MVAS) using high-speed imaging.

High-speed filming is one of the most informative methods for assessing voice physiology data. Tracing high-speed images of the glottis provides quantitative parameters such as the glottal area and the glottal width function. By way of example, a number of studies are discussed which extract quantitative data from high-speed images showing voice onsets. Furthermore, a new computer system (MVAS; multi-dimensional voice analysis system) is presented that synchronously displays a laryngoscopic high-speed film, the electroglottographical signal, and several acoustic analyses of the recorded voice sample. The automatic measurement of glottal width and glottal area from the laryngoscopic images is also provided. Looking at former studies and our analyses of voice onsets reveals a tremendous intersubject and even intrasubject variability (different prephonatory closure, different time span until full amplitude is reached, different open quotient).

Investigation of vocal fold impact stress in human subjects.

The primary purpose of the study was to explore a methodology for measuring vocal fold impact stress (SI) in awake humans, and to provide information about the general magnitude of SIs that may occur at the midpoint of the membranous vocal folds during phonation. A secondary purpose was to examine the potential use of the electroglottographic closed quotient (EGG CQ) to indirectly reflect SI. Seven male and 13 female adults were enrolled as subjects, of whom 18 had normal larynges and normal voices, 1 had nodules, and 1 had vocal fold paresis and bowing. Subjects attempted to produce 3 different voice types (pressed, normal, breathy), at 3 different pitches (low, medium, high) and 3 different loudness levels (quiet, medium, loud). For a first set of trials, only EGG data were collected. For a second set, a sensor was also introduced to the midmembranous glottis for the collection of SI data. The primary findings were that (1) endolaryngeal sensor placement was achieved during phonation trials for 17 of 20 subjects; however, grossly consistent anteroposterior positioning was accomplished, and analyzable data were obtained, for only 7 subjects; (2) SIs ranged from less than 1 kPa to about 3 kPa for those 7 subjects; and (3) no relation was detected between simultaneous CQs and SIs for individual data, although a relation was reported in a prior canine study. One possible reason for the failure to show such a relation in the present study was subtle variations in vertical as well as anteroposterior positioning of the sensor during the trials. Future studies should focus on developing a methodology for ensuring invariant 3-dimensional sensor positioning between the membranous folds, so that the stability of both SI and simultaneous CQ data can be improved.

Is a duplication of 14q32 a new recurrent chromosomal alteration in B-cell non-Hodgkin lymphoma?

Identification of clonal chromosomal abnormalities involving 14q32 and its association with specific histological subtypes of non-Hodgkin lymphoma (NHL) has provided substantial insight to the genetic events leading to the disease. However, in some cases with inferior morphology of tumor cell chromosomes, the additional segment on chromosome 14 remains unidentified by cytogenetic banding techniques alone. To elucidate the origin of the additional chromosomal segment and to correlate the newly determined alterations with histology, metaphases from 15 NHL patients with add(14)(q32) were examined using fluorescence in situ hybridization (FISH) techniques after cytogenetic analysis had been performed. We found the duplication of 14q involving the q32 region in 6 cases with a dup(14) (q32) in 4 cases and a dup(14)(q24q32) in 2 cases. In 8 cases, FISH unveiled known NHL associated translocations; a t(14;18)(q32;q21) in 4 cases, a t(11;14)(q13;q32) in 2 cases, a t(8;14)(q24;q32) and a t(9;14)(p13;q32) in 1 case each. We also noted a t(14;17)(q32;q21) in 1 case. The use of FISH was a valuable asset in determining the origin of the additional material on chromosome 14q32, and helped resolve a group of B-cell NHLs with involvement of a duplicated 14q32 region.

Rearrangements of chromosome band 1p36 in non-Hodgkin's lymphoma.

We studied 850 consecutive cases of histologically ascertained pretreatment non-Hodgkin's lymphoma with cytogenetically abnormal clones. The diagnostic karyotypes revealed that 12% of these cases exhibited structural rearrangements involving chromosome band 1p36. Here, we describe the karyotypes of 53 cases containing a 1p36 rearrangement [often involving translocations of unknown material and presented as add(1)(p36)]. We used fluorescence in situ hybridization to determine the origin of the translocation partners. We report three different recurrent translocations involving 1p36. These include der(1)t(1;1)(p36;q21) (three cases), der(1)t(1;1)(p36;q25) (three cases), and der(1)t(1;9)(p36;q13) (four cases). Using cytogenetic and fluorescence in situ hybridization analyses, we have resolved the translocation partners in 31 cases. Rearrangements of band 1p36 were found among different histopathological subtypes. Alterations of 1p36 never occurred as a sole abnormality, and in 42 of 53 cases, alterations of the band 14q32 were observed. The t(14;18)(q32;q21) translocation was present in 35 cases. The significantly high occurrence of 1p36 breakpoint in structural rearrangements and its involvement in recurrent translocations suggest that the region is bearing gene(s) that are important in lymphomagenesis. Our study also showed that cytogenetically evident deletions were frequent in chromosome 1p, almost always involving the p36 region, whereas duplications were rare and never encompassed the p36 region. Chromosome band 1p36 harbors many candidate tumor suppressor genes, and we propose that one or more of these genes might be deleted or functionally disrupted as a molecular consequence of the rearrangements, thus contributing to lymphomagenesis.

Pulmonary function after total laryngectomy.

OBJECTIVE: To present data on pulmonary function in 59 laryngectomees using a specially designed silicone adapter for connection of the stoma to the bodyplethysmograph. STUDY DESIGN: Prospective assessment of pulmonary function in 59 patients, and comparison of lung function before and after bronchodilator testing in selected cases. METHODS: The usefulness of the adapter was examined. Data of patients with airway obstruction were tabulated according to large airway obstruction (LAO), peripheral airway obstruction (PAO), and small airway disease (SAD) types. RESULTS: Findings show that pulmonary airway obstruction was present in 81% of patients and normal pulmonary function was present in only 11 of the 59 participants (19%). LAO was found in 25%, PAO in 17%, and SAD in 39% of cases. Emphysema was diagnosed in 14% of cases within the above-mentioned LAO and PAO groups. Improvement of pulmonary function was achieved in 12 of 16 laryngectomees with airway obstruction, when a bronchodilator aerosol was administered. In 60% of cases with LAO and PAO, the laryngectomees did not know of any marked obstruction of their airways, and only 10% of those knowing about their obstruction received appropriate medical treatment. CONCLUSIONS: These results suggest that 42% of the laryngectomees tested may have benefited from further medical treatment. After laryngectomy, pulmonary function assessment was performed elsewhere in 1 of 59 cases. In light of the high prevalence of airway obstruction in laryngectomees, more frequent postoperative assessments of pulmonary function should be offered to prevent or to reduce impairment of respiratory function in this postlaryngectomy vulnerable pulmonary status. Further studies are also needed to determine the effects of therapeutic intervention, e.g., assessment of therapy outcome and influence on quality of life.

Deletion of cell division cycle 2-like 1 gene locus on 1p36 in non-Hodgkin lymphoma.

Our laboratories have documented a significantly high occurrence of chromosome 1p36 rearrangements in non-Hodgkin lymphoma (NHL). The cell division cycle 2-like 1(CDC2L1) (also known as TP58 or PITSLRE) gene, a protein kinase implicated in apoptotic signaling, is located at the very distal region of chromosome 1p36 and is likely to be disrupted by structural rearrangements involving 1p36. To determine the molecular consequences of the recurrent involvement of the 1p36 region, we examined metaphases containing 1p36 abnormalities from 31 specimens derived from 26 patients for the possible deletion of CDC2L1 by fluorescence in situ hybridization (FISH) using the TP58clk-1 DNA probe. Twenty-three cases exhibited the loss of CDC2L1 from the abnormal chromosome 1. In 2 of 26 cases, the gene locus was translocated to the partner chromosome, and in four specimens, all derived from one case, CDC2L1 was not deleted. This pilot investigation suggests that 1p36 rearrangements, and consequently the loss of the CDC2L1 gene locus, is important in NHL. This work also opens avenues for further molecular studies and prognostic correlations.

Endolaryngeal contact pressures.

In this work, we present a new method for in vivo endolaryngeal contact pressure measurement with a miniature pressure transducer. Using this methodology, contact pressures can be measured during videoendoscopy at different locations between the artyenoids and also at various locations along the membranous vocal folds. Twenty adults with organic and functional voice disorders and two vocally healthy adults participated as subjects. Endolaryngeal contact pressure measures were made during a series of phonatory tasks varying pitch, loudness, and phonatory onset and offset. Measures were also made during nonphonatory tasks, including throat clearing, coughing, Valsalva maneuvres, and gagging. The most remarkable findings were: (1) interarytenoid contact pressures were considerably greater than intraglottal contact pressures; (2) interarytenoid contact pressures were greater for lower than higher pitches; (3) both interarytenoid and intraglottal contact pressures were remarkably large during hard glottal attack; and (4) overall, the largest endolaryngeal pressures were recorded between the arytenoids, during a thoracic fixation maneuver and during gag reflex.

Fluconazole pharmacokinetics in burn patients.

The pharmacokinetics of fluconazole in nine adult patients with severe (30 to 95% total body surface area) burns were studied. There was no significant difference in half-life (t1/2), clearance (CL), or volume of distribution (V) over time in five patients on days 3 and 8 of the study (P > 0.05). Combined parameter estimates (means +/- standard deviations) for all nine patients for the two study periods were as follows: t1/2, 24.4 +/- 5.8 h; CL, 0.36 +/- 0.09 ml/min/kg; and V, 0.72 +/- 0.12 liters/kg. These estimates of t1/2 and CL in burn patients were approximately 13% shorter and 30% more rapid, respectively, than the most extreme estimates reported for other populations.

Effect of acute phase response on phenytoin metabolism in neurotrauma patients.

The purpose of this prospective study was to correlate measures of the acute phase response, associated therapeutic interventions, and other clinical variables with the process of altered drug metabolism previously observed in patients with severe neurotrauma. Nine patients with severe head injury (Glasgow Coma Scale < or = 8) requiring intravenous phenytoin were included in the study. A loading dose of phenytoin was followed by daily maintenance doses. Serial blood samples were taken after the loading dose and every even-numbered study day for 10 to 14 days for measurement of total and unbound concentrations of phenytoin, interleukin-1 beta, interleukin-6 (IL-6), tumor necrosis factor alpha, alpha 1-acid-glycoprotein, C-reactive protein, and albumin. Time-invariant and time-variant Michaelis-Menten models were fit to the phenytoin concentration-time data. Protein intake was closely monitored. The mean (+/- SEM) unbound fraction of phenytoin increased from 0.17 +/- 0.02 on day 1 to 0.24 +/- 0.04 on day 10 (P < 0.05). The time-variant model was superior in describing the concentration-time data of unbound phenytoin in eight of nine patients. Mean (+/- SEM) pharmacokinetic parameter estimates for unbound phenytoin were: Vmax delta = 605 +/- 92 mg/day, VmaxB = 149 +/- 26.3 mg/day, K(ind) = 0.013 +/- 0.004 hr-1. Interleukin-6 was the only cytokine with significant concentration changes over time; it was inversely correlated with Vmax,t. Peak concentrations of interleukin-6 also proved to be inversely correlated with VmaxB. The daily amount of protein administered was significantly correlated with Vmax,t. Significant alterations in the metabolism of phenytoin occur after severe neurotrauma. The etiology of these changes is probably multifaceted. These results suggest that low initial phenytoin Vmax may be explained by the presence of interleukin-6. An increase in oxidative metabolism that correlated with nutritional protein administration was observed later in these patients.