Regulation of inflammatory and catabolic responses to IL-1ß in rat articular chondrocytes by microRNAs miR-122 and miR-451.

OBJECTIVE: miR-122 stimulates proliferation of growth plate chondrocytes whereas miR-451 stimulates terminal differentiation and matrix turnover. Here, we examined the potential of these microRNA as regulators of articular chondrocytes using an in vitro model of osteoarthritis. METHODS: miR-122 and miR-451 presence in rat articular cartilage was assessed using the anterior cruciate ligament transection model of OA. In vitro testing used first passage rat articular chondrocytes (rArCs) that were transfected with lipofectamine (Lipo) and miR-122 or miR-451 for 24-h, then treated with 10 ng/mL IL-1ß in order to mimic an osteoarthritic environment. Conditioned media were collected and MMP13, PGE2 and OA-related cytokines were measured. Matrix vesicles were collected from cell layer lysates using ultra-centrifugation. Cells were treated with miR-122 or miR-451 inhibitors to verify miR-specific effects. RESULTS: Both miR-122 and miR-451 were increased in the OA articular cartilage compared to healthy tissue; rArCs expressed both microRNAs in MVs. miR-122 prevented IL-1ß-dependent increases in MMP-13 and PGE2, whereas miR-451 significantly increased the IL-1ß effect. Multiplex data indicated that miR-122 reduced the stimulatory effect of IL-1ß on IL-1α, IL-2, Il-4, IL-6, GM-CSF, MIP-1A, RANTES and VEGF. In contrast, IL-2, IL-4, IL-6, GM-CSF, and MIP-1A were increased by miR-451 while VEGF was decreased. Inhibiting miR-122 exacerbated the response to IL-1ß indicating endogenous levels of miR-122 were present. There were no differences in MMP-13 or PGE2 with miR-451 Locked Nucleic Acid (LNA) inhibitor treatment. CONCLUSIONS: Both miRs were elevated in OA in a rat bilateral anterior cruciate ligament transection (ACLT) model. miR-122 prevented, while miR-451 exacerbated the effects of IL-1ß on rArCs.

Design of a pulse oximeter for price sensitive emerging markets.

While the global market for medical devices is located primarily in developed countries, price sensitive emerging markets comprise an attractive, underserved segment in which products need a unique set of value propositions to be competitive. A pulse oximeter was designed expressly for emerging markets, and a novel feature set was implemented to reduce the cost of ownership and improve the usability of the device. Innovations included the ability of the device to generate its own electricity, a built in sensor which cuts down on operating costs, and a graphical, symbolic user interface. These features yield an average reduction of over 75% in the device cost of ownership versus comparable pulse oximeters already on the market.

Health-related consequences of physical and sexual violence: women in the military.

OBJECTIVES: To identify differences in health-related quality of life among women veterans who were raped, physically assaulted (not in the context of rape or domestic violence), both, or neither during military service. METHODS: We did a cross-sectional telephone survey of a national sample of 558 women veterans who served in Vietnam and subsequent eras of military service. A stratified survey design selected subjects according to era of service and location. The interview included socioeconomic information, lifetime violence history, the Women's Military Environment Survey to assess women's military experiences, and the Medical Outcomes Study Short Form-36 to assess health-related quality of life. RESULTS: Five hundred thirty-seven women completed the interview. Half (48%) experienced violence during military service, including rape (30%), physical assault (35%), or both (16%). Women who were raped or dually victimized were more likely to report chronic health problems, prescription medication use for emotional problems, failure to complete college, and annual incomes less than $25,000 (P <.05). Women who were physically assaulted or raped reported significantly lower health-related quality of life (P <.05). Those who had both traumas reported the most severe impairment, comparable to women with chronic illnesses. CONCLUSION: This study suggests that the sequelae of violence against women are an important public health concern. More than a decade after rape or physical assault during military service, women reported severely decreased health-related quality of life, with limitations of physical and emotional health, educational and financial attainment, and severe, recurrent problems with work and social activities.

Topical lidocaine exaggerates laryngomalacia during flexible bronchoscopy.

In 156 infants and children referred for flexible fiberoptic bronchoscopy (FFB) we examined the larynx before and after application of 2% lidocaine solution to see what effect topical anesthesia might have on laryngeal function. All patients received midazolam and nalbuphine intravenously. Using blinded, randomly re-recorded key segments of the original video recordings of bronchoscopy, we used an empirical scoring system (Table 2) to grade signs of laryngomalacia on a scale of 0 to 8 points before and after application of the lidocaine solution. This score was based on the magnitude of collapse of the arytenoids and folding of the epiglottis during inspiration. Topical lidocaine was more likely to result in an increase in signs of laryngomalacia than in a decrease in such signs (p < 0.0001). Younger patients were more likely to have signs of laryngomalacia before topical anesthesia (odds ratio, 1.83). In a group of 10 additional patients, selected for history and physical findings consistent with laryngomalacia, application of a normal saline solution had no effect on the laryngomalacia score, but application of 2% lidocaine solution to the larynx resulted in a significant increase in this score (p < 0.001). Thus, this study has clearly demonstrated that topical anesthesia in infants and children undergoing FFB exaggerates the findings commonly associated with laryngomalacia. Because overestimation of these findings might lead to unnecessary treatment, it is important to examine the larynx and contiguous structures before applying topical anesthesia.

Insulin autoantibodies and insulin antibodies have similar binding characteristics.

Type 1 diabetes is an autoimmune disease characterized by immune-mediated destruction of the pancreatic beta cells. Insulin autoantibodies (IAAs) develop in many subjects at high risk for developing type 1 diabetes prior to onset of clinical disease and exposure to exogenous insulin, whereas insulin antibodies (IAs) commonly develop in patients treated with exogenous insulin. To investigate whether the binding characteristics of IAA and IA are similar, we measured eight different binding characteristics of IAAs from 19 insulin-naive first-degree relatives of type 1 diabetes patients and compared these to the binding characteristics of IAs from 19 type 1 diabetes patients treated with exogenous insulin. IAA and IA samples were matched for percentage insulin binding. Scatchard analysis revealed that IAAs have a two-slope representation similar to IAs-that is, two populations of antibodies, a high-affinity low-capacity population and a low-affinity high-capacity population. Binding properties of the two respective populations were found to be similar for IAAs and IAs. Sipps' equation was used to generate a Hill plot and produce an index of heterogeneity, which showed further similarity between IAAs and IAs. These results suggest that the IAAs found in subjects at increased risk for type 1 diabetes, like IAs, are likely to be polyclonal in nature. The similarities between IAAs and IAs support the hypothesis that both are induced by insulin presentation to the immune system.

Effectiveness of home versus hospital care in the routine treatment of cystic fibrosis.

Many cystic fibrosis patients with Pseudomonas lung infections receive intravenous (IV) antibiotics and chest physiotherapy (CPT) at home. Previous studies have suggested that home care, in the setting of a clinical study, is as efficacious as hospital care. This report compares the outcomes of home care with minimal supervision to outcomes of hospital care. We compared two groups of similar age and severity of lung impairment. Patients met strict definitions for home or hospital treatment (27 home care courses/33 hospital care courses). Five patients completed six courses of both home care and hospital treatment. Treatment in both groups included intravenous antibiotics and CPT. Primary outcome measures included changes in pulmonary function between the start of treatment and after 2 weeks of therapy, duration of treatment, and intervals between antibiotic courses. In hospitalized patients, forced vital capacity (FVC) increased by 17.4 +/- 3.1% (mean +/- SEM), and forced expiratory volume in one second (FEV1) increased by 23.3 +/- 4.1%, both significant at P < 0.001. The FVC and FEV1 of patients treated at home increased by 10.2 +/- 2.0% and 13.7 +/- 2.6% respectively, neither of which was a significant improvement. Similar results were found in the five patients completing both home and hospital courses. The average duration of treatment was twice as long and time between IV antibiotic courses only two-thirds as long for those treated at home compared with the hospitalized patients. Previous reports have claimed that home care in the setting of a prospective study is as efficacious as hospital care. Our experience indicates that routine home care with minimal supervision of patients is less effective than hospital care. Furthermore, home care as delivered to patients in this report increased the overall cost of care by as much as 30% because of longer and more frequent courses of antibiotic therapy.

Eliminating sleep-associated hypoxemia improves growth in infants with bronchopulmonary dysplasia.

OBJECTIVE: Infants with bronchopulmonary dysplasia (BPD) have been previously reported to have a decrease in growth velocity after stopping supplemental oxygen (SO). SO was stopped after a short-term recording (20-30 minutes) of pulse oxygen saturation (Sao2) of 92% or greater in room air. Other studies have documented that Sao2 decreases further during feedings and sleep in infants with BPD. Two questions were asked: (1) whether short-term, awake Sao2 studies would reliably predict prolonged sleep Sao2; and (2) how Sao2 sustained at 88% to 91% vs 92% or greater in room air would impact growth velocity in infants with BPD. METHODOLOGY: Short-term Sao2 studies were prospectively compared with prolonged sleep Sao2 (n = 63) and the growth velocity of infants who had SO discontinued after a prolonged sleep Sao2 recording of 88% to 91% (group 1; n = 14) versus 92% or greater (group 2; n = 34) in room air. RESULTS: Failure to maintain Sao2 at predetermined levels occurred in 18 (29%) of 63 infants during their first prolonged sleep study. There was no correlation between short-term awake Sao2 and prolonged sleep Sao2 recordings (r = .02). Body weight, height, weight for height, and rate of weight gain were similar for all study infants before SO was stopped and remained constant for group 2 infants after SO was stopped. However, group 1 infants had a significant decrease in the rate of weight gain (17.3 +/- 13.1 vs 3.7 +/- 6.1 g/kg per day), and the mean z scores for weight gain and weight for height also decreased significantly for group 1 infants. Energy intake, incidence of acute infection, hematocrit values, and medication use did not differ before or after stopping SO in either group. CONCLUSIONS: This study indicated that short-term, awake Sao2 measurements do not predict prolonged sleep Sao2, and overall, infants with BPD continued a positive growth trend when Sao2, remained greater than 92% during prolonged sleep.

Improved method for deep thoracoscopic lung nodule excision.

Minimally invasive excision of deep lung nodules can be difficult partly because endoscopic staplers do not open wide enough. We describe a method in which a standard atraumatic straight clamp compresses the lung to allow placement of the stapler without difficulty.

Concomitant coronary artery bypass grafting and adrenalectomy for pheochromocytoma.

We report the first case of concomitant coronary artery bypass grafting (CABG) and adrenalectomy for pheochromocytoma. Which lesion to correct first and staging of the corrective procedures are highly controversial issues. Issues concerning pre- and intraoperative control of hemodynamic instability from catecholamine release are discussed. Preoperative alpha and beta blockade permitted excellent hemodynamic control in the operating room. Operative and postoperative courses were uneventful and the patient had complete resolution of his angina and hypertension. We conclude that concomitant CABG and adrenalectomy is the preferred approach when pheochromocytoma and symptomatic coronary artery atherosclerosis coexist.

Changes in lung lipid during spontaneous labor in fetal sheep.

The goals of this study were 1) to examine changes in lung liquid formation and composition during spontaneous labor in fetal lambs and 2) to determine the importance of beta-adrenergic stimulation and transepithelial Na+ flux in removing liquid from the lung lumen near birth. We measured net production of lung liquid (Jv), lung liquid composition, and transpulmonary electrical potential difference (PD) before and during labor in fetal sheep with chronically implanted tracheal and vascular catheters. We determined Jv by measuring rate of change in lung liquid concentration of 125I-albumin, an impermeant tracer that was mixed in lung liquid at the start of each study. In 17 paired experiments, Jv decreased from 11 +/- 2 ml/h (Jv > 0 = secretion) before labor to -1 +/- 2 ml/h (Jv < 0 = absorption) during labor; in 5 paired experiments, PD changed from -7 +/- 1 mV (lumen negative) before labor to -12 +/- 1 mV during labor. To determine whether absorption of lung liquid during labor is the result of beta-adrenergic stimulation, we studied the effect of propranolol on Jv during labor. When propranolol (40 microM) was added to lung liquid during active labor, Jv decreased from -2 +/- 2 to -8 +/- 3 ml/h (n = 9). Thus, propranolol did not inhibit lung liquid absorption during labor. To determine whether transepithelial Na+ movement provides the driving force for lung liquid clearance during labor, we tested the effects of amiloride, an Na+ transport inhibitor, on Jv and PD.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of venous fistula technique for intraoperative cardiac hemorrhage.

We describe an innovative technique to control hemorrhage after right atrial wall reconstruction in a patient with primary angiosarcoma. At the time of surgery, the tumor was found to involve all of the free wall of the right atrium and a superficial layer of the right ventricle. A pericardial patch was used to reconstruct the right atrial wall; however, there was diffuse coagulopathy with bleeding from the anastomoses and remaining tumor. To tamponade the bleeding, a Dacron patch was sewn to the right edge of the pericardium, around the right ventricle and right coronary artery, but with little improvement in the bleeding. A type of Cabrol fistula was then formed with a tube graft from the Dacron patch to the left innominate vein. This acted as a conduit back into the venous system, with the blood taking the path of least resistance. Hemostasis was then achieved, allowing the patient to be taken to the intensive care unit.

Reversible megaesophagus associated with atypical primary hypoadrenocorticism in a dog.

Megaesophagus, hypercalcemia, and eosinophilia associated with glucocorticoid deficiency were detected in a 5-year-old neutered female Standard Poodle with concurrent hypothyroidism. Clinical and biochemical abnormalities resolved with glucocorticoid replacement treatment, and the dog was normal 29 months after diagnosis. The dog's breed and sex and the existence of a second endocrinopathy supported an underlying immunologic disorder.

Developmental changes in lung epithelial ion transport and liquid movement.

During fetal life, the lungs are filled with liquid that flows from the pulmonary circulation across the epithelium in response to the osmotic force generated by Cl- secretion of airway and distal lung epithelial cells. As birth approaches, net Cl- secretion across the respiratory tract epithelium decreases, and this is associated with a reduction in the flow of liquid into the lung lumen. The cause for this change is unknown, but several recent studies indicate that it may be related to alterations in the hormonal milieu to which the lung epithelium is exposed late in gestation. The switch from placental to pulmonary gas exchange at birth requires rapid removal of liquid from the lung lumen. During labor and the immediate postnatal period, the pulmonary epithelium changes from a predominantly Cl-(-)secreting membrane to a predominantly Na(+)-absorbing membrane, with resultant reversal of the direction of flow of lung liquid. There is considerable evidence that this change reflects an active metabolic process involving increased Na(+)-K(+)-ATPase activity in lung epithelial cells, which drives liquid from the lung lumen into the interstitium, with subsequent absorption into the pulmonary circulation. This Na(+)-K(+)-ATPase-dependent process persists in the bronchopulmonary epithelium of the mature lung and probably has an important role in clearance of alveolar edema associated with heart failure or lung injury.

Person-to-person transmission of Pseudomonas cepacia between patients with cystic fibrosis.

Ribotyping, a method of strain identification based on analysis of bacterial genomic restriction fragment length polymorphisms, was used to investigate the acquisition of Pseudomonas cepacia by a patient with cystic fibrosis. Analysis of isolates recovered from the index patient and his contacts showed person-to-person transmission of this opportunist organism. This documentation of the transmission of P cepacia from one cystic fibrosis patient to another suggests that measures to limit the acquisition of the pathogen by patients with cystic fibrosis may be worth while.

Stridor and gastroesophageal reflux in infants.

A relation was found between persistent stridor and gastroesophageal reflux in seven infants, aged 6 weeks to 6 months. Stridor began at 11 days to 2 months of age, and four of the seven infants had transient hypercarbia on at least one occasion before study. Only one had a history of frequent vomiting; three had recurrent pneumonia. Midesophageal pH, chest and abdominal movement, exhaled carbon dioxide partial pressure, and heart rate of six of the infants were recorded for 4 to 12 hours as they slept. Esophageal pH of the seventh infant was recorded for 24 hours. In the six completely studied infants, there were persistent increases of greater than 10 mm Hg in exhaled carbon dioxide level (three infants), of greater than 10 breaths per minute in respiratory rate (four infants), and in retractions and stridor (six infants) 5 to 20 minutes after onset of reflux. Stridor improved with medical management in 48 hours (five of five infants) and disappeared in 3 weeks (three of five infants) to 2 months (one of five infants). One of these medically treated infants subsequently was treated by Nissen gastric fundoplication because of a recurrence of persistent and severe stridor. Three infants had antireflux surgery, and in two of these stridor disappeared in 48 hours. In the third infant stridor disappeared 3 weeks after surgery. Based on this experience, reflux occasionally causes stridor, probably because of acute inflammation of the upper airway. If structural anomalies are ruled out, infants with severe stridor should be examined for gastroesophageal reflux.

Effects of amiloride on alveolar epithelial PD and fluid composition in rabbits.

To look for evidence of active absorption of Na+ in the alveolus in vivo in air-filled lungs, we measured [K+] and [Cl-] in the alveolar lining fluid and the potential difference (PD) across the alveolar epithelium by puncturing alveoli in lungs of anesthetized rabbits with nonselective and ion-selective microelectrodes. After intravenous doses of amiloride, the PD and [K+] decreased (-1.0 +/- 0.3 to -0.3 +/- 0.1 mV, 7.4 +/- 1.1 to 4.2 +/- 0.4 meq/l, P less than 0.001), but [Cl-] did not change (96 +/- 9, 94 +/- 4 meq/l). In another set of experiments, the PD was measured with microelectrodes filled with an electrolyte solution, and midway through each measurement some of the solution was injected into the alveolar lumen. Injecting the solution without amiloride did not alter the alveolar PD (-1.0 +/- 0.4 before and -1.1 +/- 0.5 after injection). The alveolar PD decreased to -0.1 +/- 0.2 mV after injecting the solution with 10(-5) M amiloride into the alveolar interior. These results support the hypothesis that alveolar epithelium in air-filled lungs actively absorbs sodium in vivo, which accounts for the majority of the transepithelial PD.

Calcium increases in pulmonary alveolar fluid in lambs at birth.

Calcium concentration in fetal lung fluid in lambs is relatively low, about 1.0 mEq/liter, and its concentration in alveolar fluid in mature rabbits is about 3 mEq/liter. To see if the Ca++ concentration changes at birth, we measured alveolar Ca++ as soon as possible after delivery by cesarean section and the onset of ventilation in eight anesthetized lambs at 144 days gestation. Subpleural alveoli were punctured with Ca++ selective microelectrodes as soon as 4 min and as late as 220 min after the onset of mechanical ventilation. The [Ca++] was 1.2 +/- 0.2 mEq/liter (mean +/- SD, n = 8) in fetal lung fluid collected before ventilation. After about 25 min of ventilation, alveolar Ca++ was not different from that in term lambs 24 to 72 h old (3.3 +/- 0.6 mEq/liter, n = 8). The [Ca++] increased with a t1/2 of about 10 min. Thus, alveolar Ca++ assumes a mature character very rapidly after the start of breathing in term lambs.