Activation of human monocyte-derived macrophages cultured on Teflon: response to interferon-gamma during terminal maturation in vitro

Macrophages (M phi) are potential antitumor effector cells derived from circulating blood monocytes (mo). Most studies on human mo/M phi biology and function have been performed using immature mo precursor cells. However, the conclusions drawn may be questionable, as mo have to undergo terminal differentiation before they reach relevant tissue sites of inflammation and immune reaction. We have analyzed the ability of mo-derived, teflon-cultured M phi to respond to activating stimuli with an increased tumor cytotoxic effector cell function using recombinant interferon-gamma (IFN-gamma), IFN-alpha 2, granulocyte/macrophage colony stimulating factor (GM-CSF), interleukin(IL) 2, IL 1 alpha, and bacterial lipopolysaccharides (LPS) as mediator molecules. It could be shown that the response of M phi to the most potent activator molecule, IFN-gamma, depends on the terminal differentiation from the mo stage to the mature M phi. Whereas adherent mo could be activated only moderately, M phi increased their cytotoxicity by a factor of up to 400. IFN-gamma activation positively correlated with the effector cell number, the time of incubation and the dosage used. Activation did not depend on the presence of LPS, and was lost within 24 to 48 h. LPS itself activated cells only in the microgram range. IFN-alpha 2 activated M phi only at a two log higher concentration than IFN-gamma; GM-CSF was only slightly effective, whereas M phi incubation with IL 1 alpha or IL 2 did not result in M phi activation. Thus, the ability of human M phi to become activated appears to be a function of cellular maturation and is acquired during the terminal step of M phi differentiation. Teflon-cultured M phi could facilitate studies of the activation of human M phi and may be more suitable cells for adoptive immunotherapy in cancer patients than blood monocytes.     

Identification of two mutations in a compound heterozygous child with dihydrolipoamide dehydrogenase deficiency

An infant girl with elevated blood lactate, pyruvate, and plasma branched-chain amino acids was diagnosed with dihydrolipoamide dehydrogenase (E3; dihydrolipoamide: NAD+ oxidoreductase, EC 1.8.1.4) deficiency. Activities of the pyruvate dehydrogenase complex and E3 from patient were 26 and 2% of controls in blood lymphocytes, and 11 and 14% in cultured skin fibroblasts, respectively. Western blot analysis demonstrated that the amount of E3 protein in fibroblasts from the patient and her father was about half of controls, while Northern blot analysis showed normal amounts of E3 RNA. DNA sequencing of cloned full-length E3 cDNAs from the patient revealed two mutations in separate alleles. One is a single base insertion of an extra adenine in the last codon of the leader peptide sequence (TAC-->TAAC) leading to a nonsense mutation which results in the premature termination of the precursor E3 polypeptide (Y35X). The other is a missense mutation due to substitution of guanine for adenine, causing an Arg-->Gly substitution at amino acid 460 of the mature protein (R460G) which triggers the loss of E3 activity probably by structural change in the E3 dimer. DNA sequencing of E3 cDNAs from the parents demonstrated that the nonsense mutation was inherited from the father and the missense mutation was inherited from the mother.      

Psychological stress, ego defenses, and cortisol production in children hospitalized for elective surgery.

This study was designed to investigate the relationship between the effectiveness of coping mechanisms and physiological indicators of distress in children faced with the experience of hospitalization and surgery. Twenty-five children between the ages of 7 and 11 were studied in the out-patient department, 2 weeks before surgery, and again during their hospital stay. Effectiveness of defenses and defense style was measured by a clinical interview and by the Rorschach test. Cortisol production rates were measured by the analysis of 24-hour urine collections at home and again in the hospital. Ward adjustment was also rated by a ward questionnaire. The results indicated no relationship between defense effectiveness and cortisol production rates in the out-patient department and an inverse relationship between cortisol production and defense effectiveness under the stress of hospitalization. Defense style was found to correlate with coping under stress. Four different groups of children emerged, suggesting four different types of reaction to the hospital experience.     

Biological resurfacing of the knee: a review of surgical management

Lesions of the articular surfaces of the knee have been managed by various techniques over the last 50 years. Surgical management has involved: excising the damaged area, refashioning the underlying bone to produce a fibrous response, and introducing allograft, autograft and synthetic materials to encourage a repair matrix. The techniques and their pitfalls are reviewed and discussed, and suggestions made as to the direction of future studies for the repair of osteochondral lesions in the painful knee.     

Age-dependent occurrence of an ascending axon on the omega neuron of the cricket, Teleogryllus oceanicus

The omega neurons (ON1s) are a mirror-symmetrical pair of identified prothoracic auditory interneurons of crickets which have been previously described as intraganglionic. Using intracellular techniques we stained ON1s of female Teleogryllus oceanicus and found that many ON1s have axons which project anteriorly out of the prothoracic ganglion. The ascending axon arises contralateral to the soma at the most anteriolateral bend of the bow-shaped process of an otherwise "archetypical" ON1 and travels up the neck connective in a ventral position just inside the connective tissue sheath. The occurrence of the ascending axon is age-dependent. Seventy-five percent of ON1s stained in late nymphal stages and in young adults had an ascending axon while only 30% of ON1s in older adults had an ascending axon. Evidence is presented to show that ON1s having ascending axons are developmental variants of the "archetypical" ON1 and do not represent a separate neuron type. The two morphological types of ON1s are not distinguishable on the basis of their responses to sound stimuli having carrier frequencies of 3.5-60 kHz. Although we know that the ascending axon conducts action potentials, its target and terminal morphology are not yet known.     

Efficacy and Safety of Atezolizumab Plus Bevacizumab Following Disease Progression on Atezolizumab or Sunitinib Monotherapy in Patients with Metastatic Renal Cell Carcinoma in IMmotion150: A Randomized Phase 2 Clinical Trial

BackgroundThe use of immune checkpoint inhibitors combined with vascular endothelial growth factor (VEGF)-targeted therapy as second-line treatment for metastatic clear cell renal cancer (mRCC) has not been evaluated prospectively.ObjectiveTo evaluate the efficacy and safety of atezolizumab + bevacizumab following disease progression on atezolizumab or sunitinib monotherapy in patients with mRCC.Design, setting, and participantsIMmotion150 was a multicenter, randomized, open-label, phase 2 study of patients with untreated mRCC. Patients randomized to the atezolizumab or sunitinib arm who had investigator-assessed progression as per RECIST 1.1 could be treated with second-line atezolizumab + bevacizumab.InterventionPatients received atezolizumab 1200 mg intravenously (IV) plus bevacizumab 15 mg/kg IV every 3 wk following disease progression on either atezolizumab or sunitinib monotherapy.Outcome measurements and statistical analysisThe secondary endpoints analyzed during the second-line part of IMmotion150 included objective response rate (ORR), progression-free survival (PFS), and safety. PFS was examined using Kaplan-Meier methods.Results and limitationsFifty-nine patients in the atezolizumab arm and 78 in the sunitinib arm were eligible, and 103 initiated second-line atezolizumab + bevacizumab (atezolizumab arm, n = 44; sunitinib arm, n = 59). ORR (95% confidence interval [CI]) was 27% (19–37%). The median PFS (95% CI) from the start of second line was 8.7 (5.6–13.7) mo. The median event follow-up duration was 19.4 (12.9–21.9) mo among the 25 patients without a PFS event. Eighty-six (83%) patients had treatment-related adverse events; 31 of 103 (30%) had grade 3/4 events. Limitations were the small sample size and selection for progressors.ConclusionsThe atezolizumab + bevacizumab combination had activity and was tolerable in patients with progression on atezolizumab or sunitinib. Further studies are needed to investigate sequencing strategies in mRCC.Patient summaryPatients with advanced kidney cancer whose disease had worsened during treatment with atezolizumab or sunitinib began second-line treatment with atezolizumab + bevacizumab. Tumors shrank in more than one-quarter of patients treated with this combination, and side effects were manageable.     

The Effect of State-Level Sex Education Policies on Youth Sexual Behaviors

Two types of sex education are generally offered in the U.S. abstinence-only and comprehensive sex education. There is no clear scientific consensus over which approach minimizes the risk of unintended pregnancy and sexually transmitted diseases for youth. While there have been many studies of specific programs in clinical or quasi-experimental settings, there are very few evaluations of how state-level sex education policies affect the youth population. We estimate the impact of various state-level sex education policies on youth sexual activity and contraceptive use using data from four waves of the Youth Risk Behavior Surveillance System from 39 states. We found that states that require sexuality (sex and/or HIV/STD) education and contraceptive content or states that mandate education but leave the actual content up to local districts have lower rates of sexually active youth and higher rates of contraception use when youth are sexually active. States that require sexuality education and require abstinence content increase the rate at which youth are sexually active, and youth in those states are less likely to use hormonal birth control if they are sexually active. In conclusion, we found that state policies regarding sex and HIV/STD education had statistically significant effects that are meaningful in magnitude from a public health perspective.

     

Review of dry powder inhalers

The search for alternatives to metered-dose inhalers has accelerated recently in a bid to find effective products that do not use chlorofluorocarbon (CFC) propellants. This paper reviews the factors to be considered in developing dry powder inhalers (DPIs), particularly the formulation, metering design and flow path in the device. The advantages and disadvantages of current DPIs are discussed and possible future approaches outlined.     

Eating disorders in adolescents and young women with spina bifida

This is the first report of the association of spina bifida and eating disorders. Five patients were diagnosed rather late in the course of their illness. They all had been overweight premorbidly and had been urged to lose weight for years in order to improve their mobility. As they dieted, they experienced their weight loss as a source of power that could somehow compensate for their losses and neurologic limitations. They responded to a multidisciplinary intervention. Clinicians taking care of patients with spina bifida need to be cognizant that they may be at risk of developing an eating disorder. Such awareness should influence the quality of nutritional counseling (prevention aspect) and the clinical assessment of sudden weight loss (early intervention).