Downregulation of transcription factor SOX2 in cancer stem cells suppresses growth and metastasis of lung cancer.

BACKGROUND: The cancer stem cell hypothesis suggests that neoplastic clones are maintained exclusively by a small subpopulation of cells, which have indefinite proliferation and differentiation potentials and give rise to phenotypically diverse cancer cells. Cancer stem cells have been isolated by their ability to efflux Hoechst 33342 dye and are referred to as the 'side population' (SP). METHODS AND RESULTS: The Hoechst efflux assay was used to isolate and characterize the SP from murine D121 lung carcinoma cells. Here, we demonstrated that D121-SP cells contain cancer stem cell characteristics, that is, upregulation of the transcription factors SOX2 and Oct 4 in D121-SP cells. In addition, the migration of D121-SP was decreased, and apoptosis of D121-SP was upregulated following knocking down of SOX2 in D121 cells. Importantly, downregulation of SOX2 in D121 cells markedly suppressed their metastatic potential in syngeneic mice. CONCLUSIONS: These results suggest that the SP is an enriched source of lung tumour cells with stem cell properties and that SOX2 has an important role in maintaining stem cell properties and functions that may be a potential target for effective lung cancer therapy.

The role of proto-oncogene Fra-1 in remodeling the tumor microenvironment in support of breast tumor cell invasion and progression.

A growing body of evidence indicates that interactions between neoplastic cells and tumor-associated macrophages (TAMs) in the tumor microenvironment (TME) are crucial in promoting tumor cell invasion and progression. Macrophages have an ambiguous role in these processes as this M1 phenotype correlates with tumoricidal capacity, whereas TAMs of M2 phenotype exert tumor-promoting effects. In this study, we provide evidence that interactions between mouse breast tumor cells and TAMs remodel the TME, leading to the upregulation of Fra-1, a member of the FOS family of transcription factor. In turn, this proto-oncogene initiates activation of the IL-6/JAK/Stat3 signaling pathway. This creates a malignant switch in breast tumor cells, leading to increased release of proangiogenic factors MMP-9, vascular endothelial growth factor and transforming growth factor-beta from tumor cells and intensified invasion and progression of breast cancer. Proof of the concept for the crucial role played by transcription factor Fra-1 in regulating these processes was established by specific knockdown of Fra-1 with small interfering RNA, which resulted in a marked suppression of tumor cell invasion, angiogenesis and metastasis in a mouse breast cancer model. Such a strategy could eventually lead to future efficacious treatments of metastatic breast cancer.

Pharmacodynamic analysis and clinical trial of amoxicillin sprinkle administered once daily for 7 days compared to penicillin V potassium administered four times daily for 10 days in the treatment of tonsillopharyngitis due to Streptococcus pyogenes in children.

An a priori pharmacokinetic/pharmacodynamic (PK/PD) target of 40% daily time above the MIC (T >MIC; based on the MIC(90) of 0.06 microg/ml for Streptococcus pyogenes reported in the literature) was shown to be achievable in a phase 1 study of 23 children with a once-daily (QD) modified-release, multiparticulate formulation of amoxicillin (amoxicillin sprinkle). The daily T >MIC achieved with the QD amoxicillin sprinkle formulation was comparable to that achieved with a four-times-daily (QID) penicillin VK suspension. An investigator-blinded, randomized, parallel-group, multicenter study involving 579 children 6 months to 12 years old with acute streptococcal tonsillopharyngitis was then undertaken. Children were randomly assigned 1:1 to receive either the amoxicillin sprinkle (475 mg for ages 6 months to 4 years, 775 mg for ages 5 to 12 years) QD for 7 days or 10 mg/kg of body weight of penicillin VK QID for 10 days (up to the maximum dose of 250 mg QID). Unexpectedly, the rates of bacteriological eradication at the test of cure were 65.3% (132/202) for the amoxicillin sprinkle and 68.0% (132/194) for penicillin VK (95% confidence interval, -12.0% to 6.6%). Thus, neither antibiotic regimen met the minimum criterion of > or =85% eradication ordinarily required by the U.S. FDA for first-line treatment of tonsillopharyngitis due to S. pyogenes. The results of subgroup analyses across demographic characteristics and current infection characteristics and by age/weight categories were consistent with the primary-efficacy result. The clinical cure rates for amoxicillin sprinkle and penicillin VK were 86.1% (216/251) and 91.9% (204/222), respectively (95% confidence interval, -11.6% to -0.4%). The results of a post hoc PD analysis suggested that a requirement for 60% daily T >MIC(90) more accurately predicted the observed high failure rates for bacteriologic eradication with the amoxicillin sprinkle and penicillin VK suspension studied. Based on the association between longer treatment courses and maximal bacterial eradication rates reported in the literature, an alternative composite PK/PD target taking into consideration the duration of therapy, or total T >MIC, was considered and provides an alternative explanation for the observed failure rate of amoxicillin sprinkle.

Blood pressure variability in acute ischemic stroke depends on hemispheric stroke location.

The relationship between blood pressure (BP) variability and stroke location was examined in 85 patients admitted with acute ischemic stroke. The patients were divided into three groups according to stroke location: right hemisphere (32 patients), left hemisphere (30 patients) and non-localized (23 patients). BP upon admission was 147.94/76.53 +/- 20.72/13.70 mmHg in the right hemisphere group, 151.81/76.10 +/- 25.69/16.23 mmHg in the left hemisphere and 155.23/83.41 +/- 30.45/15.74 in the non-localized group. The left hemisphere group had significantly (p < 0.01) greater variations in systolic and diastolic BP between days 2 and 3 and in systolic BP between days 3 and 4 after stroke compared with the other groups. BP in the left hemisphere group was less stable than in the other two groups. Non-localized patients without pre-existing hypertension had a significantly lower and more stable BP during the week following stroke than non-localized patients with pre-existing hypertension. Non-localized patients with pre-existing hypertension had the highest BP and showed no improvement during the week. Systolic BP tended to be higher and less stable in left hemisphere patients than in right hemisphere, whereas among non-localized ischemic stroke patients BP was higher in those who had a prior diagnosis of hypertension.

Use of subcutaneous venous access ports to treat refractory ascites.

To assess the feasibility of peritoneal ports for management of patients with cirrhotic refractory ascites, 10 ports were placed in nine patients for frequent outpatient paracentesis. Retrospective review and telephone interviews were used to assess port performance. Kaplan-Meier analysis revealed a median duration of port patency of 255 days. In 1,557 port days, four access problems prompted further interventional evaluation. Three cases of bacterial peritonitis and one catheter obstruction developed. The use of subcutaneous venous access ports to allow control of ascites by nursing personnel is a promising alternative for management of patients with refractory ascites. Additional studies are needed to determine long-term effectiveness and safety.

Exercise performance in cystic fibrosis before and after bilateral lung transplantation.

BACKGROUND: Lung transplantation improves pulmonary function and quality of life for patients with end-stage cystic fibrosis; however, a systematic evaluation of exercise performance in lung transplant recipients with cystic fibrosis has not been reported. METHODS: Ten patients with end-stage cystic fibrosis performed incremental exercise testing before and after bilateral lung transplantation; their results were compared with those of 10 age-similar healthy volunteers. Breath-by-breath measurements of gas exchange and ventilation were obtained, arterial blood was sampled each minute, and cardiac output determined at rest and peak exercise by radionuclide ventriculography. The arterial-venous O2 content difference was derived by the Fick principle. RESULTS: After transplantation, peak O2 uptake improved (31% +/- 3% vs 45% +/- 4% predicted, P = .03) but was still reduced versus normal (100% +/- 8% predicted, p < .0001). Exercise was limited by pulmonary mechanics in all patients before transplantation but in only 2 after transplantation. Compared with control subjects, the lactate threshold occurred early, both before and after transplantation. Peak exercise cardiac output and arterial O2 content were not different from normal, either before or after transplantation. In contrast, the peak exercise arterial-venous O2 content difference was markedly reduced before and after transplantation versus normal (7.1 +/- 1.2 and 9.3 +/- 0.9 vs 17.1 +/- 1.2 mL/dL, p < or = .0001 for each) and without significant improvement. CONCLUSIONS: Exercise performance in patients with end-stage cystic fibrosis improves after lung transplantation but remains well below normal. Reduced systemic O2 extraction is an important factor limiting exercise in patients with cystic fibrosis after transplantation and may also contribute to the exercise limit before transplantation.

Systemic oxygen extraction during incremental exercise in patients with severe chronic obstructive pulmonary disease.

To determine if decreased systemic oxygen (O2) extraction contributes to the exercise limit in severe chronic obstructive pulmonary disease (COPD), 40 consecutive incremental cycle ergometer exercise tests performed by such patients, from which a "log-log" lactate threshold (LT) was identified, were compared to those of 8 patients with left ventricular failure (LVF) and 10 normal controls. Pulmonary gas exchange and minute ventilation were measured continuously and arterial blood gas tensions, pH, and lactate concentrations were sampled each minute. Cardiac output (Qc) was measured by first-pass radionuclide ventriculography. The systemic O2 extraction ratio (O2ER) was calculated as arterial - mixed venous O2 content difference (CaO2 - CvO2)/CaO2. Peak exercise O2 uptake (VO2peak) was markedly reduced in both COPD and LVF [41 (3) and 42 (3)% predicted, respectively], compared to controls [89 (2)% predicted, P < 0.0001 for each]. Similarly, the LT occurred at a low percentage of predicted maximal oxygen consumption in both COPD and LVF [25 (2) and 27 (3)%] compared to normals [46 (3)%, P < 0.0001 for each]. The systemic O2ER at peak exercise was severely reduced in COPD [0.36 (0.02)] compared to the other groups [P < 0.0001 for each], for whom it was nearly identical [0.58 (0.03) vs 0.63 (0.04), LVF vs control, P > 0.05]. In the COPD group, an early LT correlated with reduced systemic O2ER at peak exercise (r = 0.64, P < 0.0001), but not with any index of systemic O2 delivery. These data suggest that lactic acidemia during exercise in patients with severe COPD is better related to abnormal systemic O2 extraction than to its delivery and contributes to the exercise limit.

Accumulation of In-111 Octreotide in brain infarcts: a case report.

Presented is a case in which there was significant accumulation of In-111 Octreotide in brain infarcts. It is likely that the accumulation was nonspecific and due to breakdown of the blood-brain barrier.

Headgear versus function regulator in the early treatment of Class II, division 1 malocclusion: a randomized clinical trial.

A prospective randomized clinical trial was conducted to evaluate the early treatment of Class II, Division 1 malocclusion in prepubertal children. Facial and occlusal changes after treatment with either a headgear or a Fränkel function regulator are reported. Molar and canine relationships, overjet, intermolar and intercanine distances were measured from casts taken every 2 months, and mounted on a SAM II articulator. Cephalometric radiographs were taken annually. The results indicate that both the headgear and function regulator were effective in correcting the malocclusion. A common mode of action of these appliances is the possibility to generate differential growth between the jaws. The extent and nature of this effect, as well as other skeletal and occlusal responses differ. Treatment in late childhood was as effective as that in midchildhood. This finding suggests that timing of treatment in developing malocclusions may be optimal in the late mixed dentition, thus avoiding a retention phase before a later stage of orthodontic treatment with fixed appliances. However, a number of conditions may dictate an earlier intervention in the individual patient.

Escalating interferon-alpha-2b dose for patients with chronic hepatitis C.

BACKGROUND/AIMS: We examined the effectiveness of escalating the dose of interferon-alpha-2b in subjects with chronic hepatitis C who did not respond to usual treatment with 3,000,000 units 3 times a week. METHODOLOGY: Treatment was started with 3,000,000 units of interferon-alpha-2b 3 times a week. If serum alanine aminotransferase activity was not normal at 12 weeks, the dose was increased to 3,000,000 units daily. If serum alanine aminotransferase activity was not normal after 12 weeks, the dose was increased to 5,000,000 units daily. RESULTS: Fifty-one subjects started treatment. Twenty-nine subjects had their dose increased to 3,000,000 units daily and only 1 responded (3%, 95% confidence interval 0-10.9%) while 41% (95% confidence interval 21.4-60.6%) had to discontinue treatment at this dose because of adverse events or intolerance. Of 14 subjects who had their dose increased to 5,000,000 units daily, none (95% confidence interval 0-3.6%) responded, while 43% (95% confidence interval 13.5-72.5%) had to discontinue treatment. CONCLUSIONS: Escalating doses of interferon-alpha-2b are not effective and are associated with increased toxicity and intolerance in patients with chronic hepatitis who do not respond to initial treatment with 3,000,000 units 2 times a week.

Updated analysis of an outpatient chemoimmunotherapy regimen for treating metastatic melanoma.

PURPOSE: Aggressive inpatient chemoimmunotherapy protocols for metastatic melanoma have yielded encouraging response rates but have required lengthy hospitalizations. To reduce or eliminate the need for hospitalization, we have developed an outpatient chemoimmunotherapy regimen and assessed its efficacy and toxicity in 53 patients treated at the University of Washington Medical Center. PATIENTS AND METHODS: Eligible patients with measurable metastatic melanoma received carmustine (150 mg/m2 every 6-8 weeks) and dacarbazine (660 mg/m2) and cisplatin (75 mg/m2) every 3 to 4 weeks in an infusion center plus tamoxifen (20 mg/day). Patients self-administered subcutaneous recombinant interleukin-2 (rIL-2) at 3 MIU/m2/day on days 3 to 9, and recombinant interferon alfa-2a (rIFN-alpha 2a) at 3 MIU on day 3 and at 5 MIU/m2/day on days 5, 7, and 9. Maintenance rIFN-alpha 2a was self-administered subcutaneously at 5 MIU/m2 tiw for 12 months after complete or stable partial response. Response and survival were assessed. RESULTS: Fifty-three patients (median age = 49 years) have received 181 cycles. To date, there have been 10 complete responses (19%) lasting 2 to 28+ months and 12 partial responses (23%) lasting 2 to 11 months, for an overall response rate of 42% (95% confidence interval, 28%-55%). The median overall survival was 12 months. Grade 3/4 vomiting occurred in 32% of cycles, but hospitalization for supplemental intravenous fluids was required in only 11% of cycles for a median of 3 days. Grade 4 thrombocytopenia and neutropenia occurred in 9% and 8% of cycles, respectively. Grade 3 renal dysfunction occurred in only one cycle and was reversible. CONCLUSION: A chemoimmunotherapy regimen for patients with metastatic melanoma has been defined that is well tolerated on an outpatient basis and is associated with a median survival comparable to that with aggressive inpatient chemoimmunotherapy regimens.

Metastatic renal cell carcinoma: long-term survival after therapy with high-dose continuous-infusion interleukin-2.

PURPOSE: This article undertakes to define the response rate, long-term survival, and toxicity in patients with metastatic renal cell carcinoma (MRCC) treated with high-dose continuous intravenous infusion (CIV) recombinant interleukin-2 (rIL-2) with or without lymphokine-activated killer (LAK) cells. PATIENTS AND METHODS: One hundred twenty-three consecutive patients received CIV rIL-2 (18-22 MIU/m2/day on days 1-5, and 6-8 MIU/m2/day on days 10-19) on one of five sequential protocols at the University of Washington between 1988 and 1995. The first 76 patients received LAK cells. The median age was 55 years (range, 32-76 years), and 71% had undergone prior nephrectomy. RESULTS: Nine patients achieved a complete response (7.3%) and 14 patients achieved a partial response (11.4%) for an overall response rate of 19% (95% confidence interval, 12%-26%). The median survival was 19 months, and the 5-year survival was 20%. Seven of nine complete responders (78%) remain in continuing complete response at 43+ to 109+ months. Intensive care unit and vasopressor support were required in 42% and 23% of patients, respectively, who received rIL-2 + LAK cells, and in 18% and 4% of those who received rIL-2 alone. There was one treatment-related death. CONCLUSION: We report the largest single-institution experience and the longest survival for patients with MRCC treated with CIV rIL-2. The administration of rIL-2 by CIV is associated with less frequent intensive care unit and vasopressor support than with high-dose intravenous bolus regimens, and hence may enhance the therapeutic index in patients with MRCC.

Clinical and biochemical heterogeneity in females of a large pedigree with ornithine transcarbamylase deficiency due to the R141Q mutation.

A large family with ornithine transcarbamylase deficiency due to mutation R141Q was ascertained through a propositus who presented with acute neonatal hyperammonemic coma. Of 13 females at risk, 11 were evaluated clinically and had laboratory studies performed. Seven were found to be heterozygous for the mutation. Of these seven, five had chronic clinical symptoms and two were asymptomatic. None of the heterozygotes had elevated plasma ammonia on random testing. Of the five symptomatic females, three had markedly elevated plasma glutamine levels on random testing, while two had levels in the upper range of normal. Plasma citrulline and arginine levels were somewhat lower in the symptomatic individuals but still within the normal range. Five heterozygotes who were tested had either spontaneous orotic aciduria or elevated orotic acid following ingestion of allopurinol, whereas one unaffected female and one unaffected male had normal allopurinol tests. A higher than expected proportion of female heterozygous for the R141Q mutation were clinically and biochemically symptomatic but remained undiagnosed for many years. Plasma glutamine determination and allopurinol testing should be performed in females who present with a combination of relatively non-specific symptoms detailed in this report.

Gastrointestinal consequences of left ventricular assist device placement.

Left ventricular assist devices effectively improve hemodynamic function and reverse renal and hepatic dysfunction; however, their effects upon the gastrointestinal (Gl) system have not been addressed. We evaluated Gl function in 27 left ventricular assist device recipients using interviews, Gl contrast studies, endoscopy, and 99mTc sulfur colloid studies of esophageal transit and gastric emptying. While on left ventricular assist device support (mean duration of 84 days), 19 patients reported early satiety and/or nausea, and 1 was unable to tolerate oral intake. Esophageal transit time (normal, < 10 sec) was borderline slow at 14 +/- 4 (mean +/- standard error of the mean) and gastric emptying (normal < 90 min) was prolonged (range of 106-506 min, mean = 283 +/- 69 min). In a 1-38 month follow-up, gastric function subjectively improved in all. Six patients had intraperitoneal device placement. One died of aspiration pneumonia secondary to small bowel obstruction, and one had prolonged inability to tolerate oral intake, which required feeding jejunostomy tube placement. The 21 patients with pre peritoneal placement of the device did not require Gl operative interventions and had no catastrophic Gl events; they had mild to no Gl complaints. Pre peritoneal placement may mitigate early satiety and obviate serious Gl complications.

Gastroparesis after lung transplantation. Potential role in postoperative respiratory complications.

BACKGROUND: We observed an unexpectedly high incidence of postoperative gastroparesis among lung and heart-lung transplant recipients. PURPOSE: To identify the incidence of GI complications and to describe the clinical profiles of patients who developed symptomatic gastroparesis after lung transplantation. PATIENTS AND METHODS: Retrospective study of GI symptoms and complications identified during 3 years of follow-up of 38 adult lung and heart-lung transplant recipients. RESULTS: Sixteen of 38 patients (42%) reported one or more GI complaint and received a specific GI diagnosis. Nine of 38 patients (24%) complained of early satiety, epigastric fullness, anorexia, nausea, or vomiting. Gastroparesis was suspected when endoscopic evaluation revealed undigested food in the stomach after overnight fast and symptoms could not be attributed to peptide disease or cytomegalovirus gastritis. Delayed gastric emptying was confirmed by gastric scintigraphy. Mean gastric empty (t1/2) was 263 +/- 115 min (normal < 95 min). Gastroparesis occurred in 4 of 13 right lung, 2 of 12 left lung, 1 of 9 bilateral single lung, and 2 of 4 heart-lung recipients (p = NS). Patients responded partially to metoclopramide or cisapride, with the exception of two patients who required placement of jejunal feeding tubes secondary to severe symptoms. In long-term follow-up, symptoms resolved in all patients and treatment with medications or mechanical intervention was successfully discontinued. Four of nine patients (44%) suffering from gastroparesis developed obliterative bronchiolitis (OB). Food particles were discovered in the BAL fluid of two such symptomatic patients. In contrast, only 6 of 29 (21%) nonsymptomatic patients developed OB (p = 0.16). CONCLUSION: Symptomatic gastroparesis is a frequent complication of lung or heart-lung transplantation that may promote microaspiration into the lung allograft.

Monitoring growth during orthodontic treatment.

The relationship between somatic growth and orthodontic treatment has been limited to the evaluation of body height and skeletal age relative to craniofacial development. The aim of this study was to evaluate the correlation of anthropometric and biochemical measures of general growth with facial and occlusal changes during the early treatment of Class II Division 1 malocclusion. Findings are reported from 46 children, ages 7.20 to 12.85 years (skeletal ages, 5.75 to 12.75 years), who are enrolled in a prospective clinical trial. Body and knee heights were measured monthly, with a Holtain stadiometer and a Knee Height Measuring Device, respectively. Every three months, serum levels were measured of the hormone dehydroepiandrosterone sulfate (DHEAS), an androgen associated with growth in midchildhood, and osteocalcin, an indicator of bone turnover. Significant correlations existed between knee height and various occlusal measurements, but mandibular length was not significantly correlated with knee height and DHEAS levels. Knee height correlated significantly (P < .05) with DHEAS and osteocalcin only in 46% and 37% of the children, respectively. The results indicate that the evaluated biochemical measures, at the time intervals considered, may not increase the accuracy of growth depiction by physical measures alone (height and skeletal maturation).

High level expression and export of beta-glucuronidase from murine mucopolysaccharidosis VII cells corrected by a double-copy retrovirus vector.

Retrovirus vectors were constructed to transfer and express the cDNA of the human lysosomal acid hydrolase beta-glucuronidase (GUSB) under control of the human GUSB promoter. Expression of the transcription unit (minigene) was evaluated in a GUSB-negative cell line established from a mouse with the lysosomal storage disease mucopolysaccharidosis (MPS) type VII. A vector designed to transfer single copies of the minigene (N2H beta H) expressed normal levels of GUSB activity in the deficient cells. GUSB expression was increased to several times greater than normal by inserting the minigene into a double-copy vector (DCH beta H), which places one copy of the transcription unit upstream of the retrovirus promoter in both the 3' and 5' long terminal repeats (LTRs) of the integrated provirus. The specific activity of GUSB and a control normal lysosomal enzyme, alpha-galactosidase (GLA), were higher in normal and in vector-corrected cells from confluent cultures than in subconfluent dividing cells. The ratios of GUSB to GLA were similar at all phases of cell growth, but the level of GUSB expression from the double copy vector was several-fold higher than from the single copy vector. To determine if this effect was controlled by the GUSB promoter, a vector was constructed using the thymidine kinase (TK) promoter to drive the human GUSB cDNA (NTK beta H). The levels of GUSB in cells corrected with this vector exhibited the same cell density dependent pattern as when the GUSB promoter was used, indicating that the variation in enzymatic activity was not a function of the GUSB promoter.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes of arch width in the early treatment of Class II, division 1 malocclusions.

Changes in arch width during the early correction of Class II, Division 1 malocclusions with either the Fränkel functional appliance or headgear are compared in an ongoing prospective randomized clinical trial. The data were collected from 43 children, ages 7.5 to 12.85 years, who met strict dental and cephalometric criteria for inclusion in the study. They were assigned at random to treatment with either a headgear (n = 21) or a Fränkel appliance (n = 22). Occlusal measurements included the maxillary and mandibular intermolar distances (buccal and palatal/lingual) and intercanine distances. Measurements (millimeters) were performed on casts taken every 2 months, with digital calipers accurate to 0.01 mm. Four months after the initiation of treatment, the mean maxillary intermolar distance was larger in the Fränkel group (palatal: 1.58, SE: 0.22; buccal: 1.58, SE: 0.20) than the headgear group (palatal: -0.39, SE: 0.21; buccal: 0.26, SE: 0.23), and the difference was statistically significant (palatal: p < 0.0001 and buccal: p = 0.0001). The mean maxillary intercanine distance increased more with the headgear (1.62, SE: 0.19) than the Fränkel appliance (0.62, SE: 0.23) p = 0.003. As treatment progressed, the average intermolar distance in the headgear group increased, but was still higher in the Fränkel group by more than 1 mm. The intercanine distance remained larger in the headgear group. The mandibular intermolar and intercanine distances were higher after Fränkel therapy than with headgear.(ABSTRACT TRUNCATED AT 250 WORDS)