Desmoplastic melanoma: a 12-year experience with sentinel lymph node biopsy.

AIMS: Given the paucity of data regarding nodal involvement in desmoplastic melanoma (DM), we decided to review the incidence of nodal metastasis in our patients with DM to better define guidelines regarding the performance of sentinel lymph node biopsy (SLNB) in this specific melanoma subtype. METHODS: Using a prospectively maintained database, we reviewed all patients who underwent treatment for melanoma at the Yale Melanoma Unit in a twelve-year period (1998-2010), during which 3531 cases were treated. We identified 24 patients (0.7%) diagnosed with DM. These patients' records were studied for clinical and histologic parameters and clinical outcomes. RESULTS: Twenty-two patients from the DM group had SLNB, of which four (18%) were diagnosed with micro-metastasis. These four patients were all treated with completion lymphadenectomy and none had additional positive nodes in the remainder of the nodes. Patients were followed after surgery for a median of 25 months (range 2-60 months). Two patients (9%) developed local recurrence, two (9%) in-transit recurrence, and six (27%) showed distant metastases (three patients were pure DM and three patients showed mixed morphology). Patients with mixed DM had a higher rate of nodal metastasis (25%) vs those with pure DM (14%). CONCLUSIONS: Other authors have reported that patients diagnosed with pure DM were less likely to have a positive SLN (0-2%) than those patients with the mixed DM subtype (12-16%). Our findings of higher incidence rates of regional lymph node metastases in both the pure and mixed DM subtypes (14% and 25%) compel us to continue to still recommend that SLNB be considered in patients with both subcategories, pure and mixed DM. LEVEL OF EVIDENCE: Level IV.

Influence of radioactivity on surface interaction forces.

Although some differences have been observed, the transport behavior of radioactive aerosol particles has often been assumed to be analogous to the behavior of nonradioactive aerosols in dispersion models. However, radioactive particles can become electrostatically charged as a result of the decay process. Theories have been proposed to describe this self-charging phenomenon, which may have a significant effect on how these particles interact with one another and with charged surfaces in the environment. In this study, atomic force microscopy (AFM) was employed to quantify surface forces between a particle and a planar surface and to compare measurements with and without the involvement of radioactivity. The main objective of this work is to assess directly the effects of radioactivity on the surface interactions of radioactive aerosols via the measurement of the adhesion force. The adhesion force between a silicon nitride AFM tip and an activated gold substrate was measured so that any possible effects due to radioactivity could be observed. The adhesion force between the tip and the gold surface increased significantly when the gold substrate (25 mm(2) surface area) was activated to a level of approximately 0.6 mCi. The results of this investigation will prompt further work into the effects of radioactivity in particle-surface interactions.

International collaboration between US and Thailand on a clinical trial of treatment for HIV-associated cryptococcal meningitis.

BACKGROUND: International clinical trials can provide scientific and logistic benefits in spite of the many challenges. Determining whether a country, especially a developing country, is an appropriate location for the research should include in-country consultation and partnering to assess its social value for the population; that treatments are relevant for the population under study; and that the research infrastructure and ethical oversight are adequate. Collaboration increases the likelihood of study success and helps ensure that benefits accrue to recruited populations and their community. PURPOSE: This paper describes our experiences on a bi-national study and may provide guidance for those planning to engage in future collaborations. METHODS: A Thai and United States team collaborated to develop and implement a phase II clinical trial for HIV-associated cryptococcal meningitis to assess safety and tolerability of combination therapy vs. standard treatment. Clinical and cultural differences, regulatory hurdles and operational issues were addressed before and during the study to ensure a successful collaboration between the 2 groups. RESULTS: The international multicenter study allowed for more rapid enrollment, reduced costs to complete the study, sharing of the benefits of research, greater generalizability of results and capacity building in Thailand; quality metrics in Thailand were equivalent to or better than those in the U.S. CONCLUSIONS: Conducting successful clinical trials internationally requires early and ongoing collaboration to ensure the study meets sites' requirements and expectations, conforms to varying national regulations, adheres to data quality standards and is responsive to the health needs of studied populations.

Comparing stimulation-induced pain during percutaneous (intramuscular) and transcutaneous neuromuscular electric stimulation for treating shoulder subluxation in hemiplegia.

OBJECTIVE: To determine whether percutaneous (intramuscular) neuromuscular electric stimulation (perc-NMES) is less painful than transcutaneous neuromuscular electric stimulation (trans-NMES) for treating shoulder subluxation in hemiplegia. DESIGN: Double-blind, crossover trial. SETTING: University-affiliated tertiary care hospital. PARTICIPANTS: A convenience sample of 10 hemiplegic subjects with at least 1 fingerbreadth of glenohumeral subluxation. INTERVENTIONS: All subjects received 3 randomly ordered pairs of perc-NMES and trans-NMES to the supraspinatus and posterior deltoid muscles of the subluxated shoulder. Both types of stimulation were optimized to provide full joint reduction with minimal discomfort. MAIN OUTCOME MEASURES: Pain was assessed after each stimulation with a 10-cm visual analog scale (VAS) and the McGill Pain Questionnaire, using the Pain Rating Index (PRI) scoring method. Subjects were asked which type of stimulation they would prefer for 6 weeks of treatment. Wilcoxon's signed-rank test was used to compare median differences in VAS and PRI between perc-NMES and trans-NMES. RESULTS: Median VAS scores for perc-NMES and trans-NMES were 1 and 5.7, respectively (p = .007). Median PRI scores for perc-NMES and trans-NMES were 7 and 19.5, respectively (p = .018). Nine of the 10 subjects preferred perc-NMES to trans-NMES for treatment. CONCLUSION: Data suggest that perc-NMES is less painful than trans-NMES in the treatment of shoulder subluxation in hemiplegia.

Percutaneous intramuscular neuromuscular electric stimulation for the treatment of shoulder subluxation and pain in patients with chronic hemiplegia: a pilot study.

OBJECTIVE: To investigate the feasibility of percutaneous intramuscular neuromuscular electric stimulation (perc-NMES) for treating shoulder subluxation and pain in patients with chronic hemiplegia. DESIGN: Before-after trial. SETTING: University-affiliated tertiary care hospital. PARTICIPANTS: A convenience sample of 8 neurologically stable subjects with chronic hemiplegia and shoulder subluxation. INTERVENTION: Six weeks of perc-NMES to the subluxated shoulder. MAIN OUTCOME MEASURES: Shoulder subluxation (radiograph), shoulder pain (Brief Pain Inventory), motor impairment (Fugl-Meyer score), shoulder pain-free external rotation (handheld goniometer), and disability (FIM instrument) were assessed before treatment (T1), after 6 weeks of neuromuscular stimulation (T2), and at 3-month follow-up (T3). A 1-way, repeated-measures analysis of variance using the generalized estimating equation approach was used to evaluate differences from T1 to T2 and from T1 to T3 for all outcome measures. RESULTS: Subluxation (p =.0117), pain (p =.0115), shoulder pain-free external rotation (p <.0001), and disability (p =.0044) improved significantly from T1 to T2. Subluxation (p =.0066), pain (p =.0136), motor impairment (p <.0001), shoulder pain-free external rotation (p =.0234), and disability (p =.0152) improved significantly from T1 to T3. CONCLUSIONS: Perc-NMES is feasible for treating shoulder dysfunction in hemiplegia and may reduce shoulder subluxation, reduce pain, improve range of motion, enhance motor recovery, and reduce disability in patients with chronic hemiplegia and shoulder subluxation. Further investigation is warranted.

Functional expression, purification, and characterization of recombinant human pyruvate carboxylase.

The cDNA-encoding human pyruvate carboxylase (hPC) has been assembled and cloned into a very high efficiency mammalian expression vector and the construct transfected into 293T kidney cells. Stable clones expressing very high levels of hPC were produced and used as a source of the enzyme. Purification of the recombinant hPC was performed by selective precipitation with 40% ammonium sulfate followed by a single step avidin affinity chromatography, with an overall yield of 20%. Recombinant hPC purified by this method yielded a single band on SDS-PAGE with a specific activity of 20 U/mg. Kinetic analysis demonstrated that the recombinant human PC has the same properties as the native enzyme isolated from liver autopsy. This is the first report of production and purification of recombinant PC.

An assessment of two training interventions designed to increase the knowledge of obstetrical nurses and nurse-midwives about the maternal serum triple screen.

The purpose of this study was to assess the effects of two brief training interventions designed to improve nurses' and nurse-midwives' knowledge about the maternal serum triple screen. The low intervention consisted of written information on the triple screen; the high intervention consisted of written information plus a one hour oral presentation. Knowledge was assessed at baseline, immediately following the oral presentation (high intervention only), and one month following the interventions. Forty-seven nurses, nurse-midwives and nursing assistants participated. Sixteen respondents (34 per cent) who routinely talk to patients about the triple screen obtained a score of less than 70 per cent on the knowledge questionnaire at baseline assessment. Respondents' knowledge about the maternal serum triple screen included areas that needed to be improved in order for them to be able to provide patients with accurate and complete information. Both interventions assessed in this study resulted in an increase in participants' knowledge about the maternal serum triple screen, however the high intervention was more effective. This study presents evidence that improvements in health care professionals' knowledge can be made with brief educational interventions.

Prenatal diagnosis of ring chromosome 6 in a fetus with hydrocephalus.

A patient with ring chromosome 6/monosomy 6 mosaicism is presented. At 25 weeks' gestation, ultrasound examination demonstrated fetal hydrocephalus. Amniocentesis was performed. The fetal karyotype was 45,XY,-6/ 45,XY,-6,+f/46,XY,r(6)(p25q27). Delivery of this male infant was by Caesarean section at 37 weeks' gestation. The karyotype in peripheral blood lymphocytes was 46,XY,r(6)(p25q27) with no indications of mosaicism. The infant had hydrocephalus which required treatment with a ventriculoperitoneal shunt at 22 days of age. He had no other obvious serious congenital anomalies. By 17 months he had developed microcephaly, seizures, severe bilateral hearing loss, and global development delay. This patient provides information regarding phenotypic variability of ring chromosome 6 and also reinforces the importance of offering amniocentesis if fetal hydrocephalus is detected as an isolated anomaly.

Identification of novel alternatively spliced pyruvate carboxylase mRNAs with divergent 5'-untranslated regions which are expressed in a tissue-specific manner.

We have identified and characterized multiple mRNA transcripts of rat and human pyruvate carboxylases [EC 6.4.1.1] using rapid amplification of cDNA 5' ends-polymerase chain reaction (RACE-PCR). Five alternative forms of rat pyruvate carboxylase cDNAs have been identified in liver, kidney, brain, and adipose tissue and these are expressed in a tissue-specific manner. Two alternative forms of human pyruvate carboxylase cDNA have also been identified in liver. These pyruvate carboxylase cDNAs have a common coding region but differ in their 5' untranslated regions (5'UTRs), suggesting that they are generated by alternative splicing of the primary transcript. Southern blot analysis of restriction enzyme digested rat genomic DNA revealed that pyruvate carboxylase is encoded by a single copy gene.

Polymorphism of the yeast pyruvate carboxylase 2 gene and protein: effects on protein biotinylation.

In Saccharomyces cerevisiae there are two isoenzymes of pyruvate carboxylase (Pyc) encoded by separate genes designated PYC1 and PYC2. We report the isolation and sequencing of a PYC2 gene, and the localization of both genes on the physical map of S. cerevisiae. Comparison with the previously reported sequence [Stucka, Dequin, Salmon and Gancedo (1991) Mol. Gen. Genet. 229, 307-315] revealed significant differences within the open reading frame. The most notable difference was near the 3' end, where we found a single base deletion reducing the open reading frame by 15 bases. We have confirmed the C-terminus of Pyc2 encoded by the gene isolated here by expressing and purifying an 86-amino-acid biotin-domain peptide. In addition, we investigated the effects of the two changes in the Pyc2 biotin domain (K1155R substitution and Q1178P/five-amino-acid extension) on the extent of biotinylation in vivo by Escherichia coli biotin ligase, and compared the biotinylation of peptides containing these changes with that of two different-length Pyc1 biotin-domain peptides. The K1155R substitution had very little effect on biotinylation, but the five-amino-acid C-terminal extension to Pyc2 and the N-terminal extension to Pycl both improved biotinylation in vivo.

A new hospital library: a marketing opportunity.

A new or remodeled library presents a unique marketing opportunity for the hospital librarian. Furthermore, a well-designed library markets itself through its convenience, attractiveness, and ease of use. A marketing approach to library planning takes into account needs of users and of library staff and considers the librarian's relations with the architect as well as with hospital employees. This paper describes ways to combine library planning with marketing techniques and specifies aspects of the library that contribute to its good image.

Assignment of the human pyruvate carboxylase gene (PC) to 11q13.4 by fluorescence in situ hybridisation.

The gene for pyruvate carboxylase (PC) has been localised by fluorescence in situ hybridisation to normal and FRA11A human chromosomes. PC maps to 11q13 and distal to FRA11A, localising PC to 11q13.4-->q13.5.

Designing the smaller library: a hospital looks to the future.

Like other libraries, hospital libraries are preparing for a future which will include a mix, as yet undetermined, of print and electronic information. This case study relates how the library at St. Jude Children's Research Hospital was designed to meet the needs of users and staff and to balance them with the need for more sophisticated methods of retrieving, storing, and delivering information. Details are provided on planning process and on design features such as lighting and color.

Health and well-being of childbearing women in rural and urban contexts.

As part of the national focus on women's health issues, it is important to identify those health-related characteristics of rural women that distinguish them from women living in urban settings. The aim of this study was to compare rural and urban childbearing women on socioeconomic characteristics, perceived stress, health-related practices, illness symptoms, parenting confidence, and body weight. One hundred sixty-five midwestern women responded to a health survey sent to them six months after childbirth. Rural women were younger and less educationally and economically advantaged compared to urban women. Before adjusting for these differences, rural women were less self-actualized, more interpersonally isolated, and reported less healthy nutrition than urban women. These differences disappeared when socioeconomic differences were adjusted. Rural and urban mothers did not differ in most other areas, including perceived stress, parenting confidence, and body weight. Compared to national norms, the perceived stress levels of both rural and urban mothers were significantly higher than a probability sample of U.S. women. Findings support the role of socioeconomic factors as contributing to risk of poor health promotion among rural childbearing women.

Regulation of pyruvate carboxylase isozyme (PYC1, PYC2) gene expression in Saccharomyces cerevisiae during fermentative and nonfermentative growth.

In Saccharomyces cerevisiae there are two isoenzymes of pyruvate carboxylase (Pyc) encoded by separate genes, designated PYC1 and PYC2. In the wild type yeast, the expression of both genes is influenced by both the growth phase and the type of carbon source, indicating discrete regulatory mechanisms and metabolic roles for PYC1 and PYC2. On glucose minimal medium PYC1 and PYC2 are differentially regulated as shown by a constant level of PYC1 expression throughout the main growth phase compared to a high level of PYC2 expression only in the early growth phase. On ethanol minimal medium, the growth-related pattern of PYC1 and PYC2 expression was similar as shown by a 3.6-fold decline from early to mid log phase. PYC1 expression, however, was activated 10-fold above PYC2 mRNA levels during this period of growth. To further investigate the roles of the two PYC genes we determined the growth phenotypes and expression levels of PYC in pyc1 and pyc2 single null mutants. During fermentative growth, the lack of either PYC gene had little effect on the level and pattern of expression of the other PYC gene, indicating further their separate regulation. In comparison to the pyc2 null, the pyc1 null strain showed a 3- to 4-fold lower level of Pyc activity and Pyc protein concentration. Moreover, the pyc1 null showed a strong requirement for L-aspartate for efficient growth, indicating the importance of PYC1 expression for the synthesis of C4 intermediates. DV6.2 (PYC1, pyc2 delta) showed a 3.2-fold higher level of activity on ethanol minimal medium when compared to growth on glucose minimal medium, and supported growth in the absence of L-aspartate. The pyc1 null, MW21.3 (pyc1 delta, PYC2), on the other hand, did not support growth on ethanol in the absence of aspartate. This study represents the first report on the characterisation of expression of the PYC genes in yeast throughout growth. Their metabolic roles for both fermentative and gluconeogenic growth are considered.

Rhinometry and open-mouth posture in young children.

A biracial sample of 348 elementary school children who were in the first grade were assessed for open-mouth posture (OMP) in the natural environment. In addition, rhinometry was performed on 296 of the children. Means were computed for percent OMP and cross-sectional nasal airway. Results indicated that, in general, these children exhibited relatively high rates of OMP. Boys displayed significantly greater OMP than girls, and black children showed significantly larger cross-sectional nasal areas than white children. A significant correlation between OMP and nasal area only was evident for children exhibiting OMP during more than 80% of the observation intervals. The implications of the findings were discussed.

Intratracheal aerosolization of endotoxin in the rat: a model of the adult respiratory distress syndrome (ARDS).

A technique is described for the intratracheal aerosolization of endotoxin into the rat. Using a miniaturized nozzle within the tracheal lumen to optimize uniform distribution 0.5 ml of an endotoxin solution (7 mg/kg) was aerosolized and dispersed throughout the lung. Time course studies of pulmonary function and histological changes revealed marked functional and morphological changes by 24 h. Histopathologic changes consisted of widespread pulmonary oedema and a diffuse neutrophilic alveolitis. At the same time, there were significant decreases in tidal volume, minute ventilation and lung compliance. Haematologic changes were also seen, including profound thrombocytopaenia and leukopaenia together with an increased haematocrit, indicating systemic effects in this model. Bronchoalveolar lavage (BAL) at 24 h revealed significant increases in BAL protein, erythrocytes and neutrophils. The functional, cytological and histological changes observed after endotoxin challenge mimic those seen in the Adult Respiratory Distress Syndrome in humans and can thus be used as a model to compare the efficacy of a variety of therapeutic interventions for this syndrome.