Characterization of immune responses to anti-PD-1 mono and combination immunotherapy in hematopoietic humanized mice implanted with tumor xenografts.

BACKGROUND: The success of agents that reverse T-cell inhibitory signals, such as anti-PD-1/PD-L1 therapies, has reinvigorated cancer immunotherapy research. However, since only a minority of patients respond to single-agent therapies, methods to test the potential anti-tumor activity of rational combination therapies are still needed. Conventional murine xenograft models have been hampered by their immune-compromised status; thus, we developed a hematopoietic humanized mouse model, hu-CB-BRGS, and used it to study anti-tumor human immune responses to triple-negative breast cancer (TNBC) cell line and patient-derived colorectal cancer (CRC) xenografts (PDX). METHODS: BALB/c-Rag2nullIl2rγnullSIRPαNOD (BRGS) pups were humanized through transplantation of cord blood (CB)-derived CD34+ cells. Mice were evaluated for human chimerism in the blood and assigned into experimental untreated or nivolumab groups based on chimerism. TNBC cell lines or tumor tissue from established CRC PDX models were implanted into both flanks of humanized mice and treatments ensued once tumors reached a volume of ~150mm3. Tumors were measured twice weekly. At end of study, immune organs and tumors were collected for immunological assessment. RESULTS: Humanized PDX models were successfully established with a high frequency of tumor engraftment. Humanized mice treated with anti-PD-1 exhibited increased anti-tumor human T-cell responses coupled with decreased Treg and myeloid populations that correlated with tumor growth inhibition. Combination therapies with anti-PD-1 treatment in TNBC-bearing mice reduced tumor growth in multi-drug cohorts. Finally, as observed in human colorectal patients, anti-PD-1 therapy had a strong response to a microsatellite-high CRC PDX that correlated with a higher number of human CD8+ IFNγ+ T cells in the tumor. CONCLUSION: Hu-CB-BRGS mice represent an in vivo model to study immune checkpoint blockade to human tumors. The human immune system in the mice is inherently suppressed, similar to a tumor microenvironment, and thus allows growth of human tumors. However, the suppression can be released by anti-PD-1 therapies and inhibit tumor growth of some tumors. The model offers ample access to lymph and tumor cells for in-depth immunological analysis. The tumor growth inhibition correlates with increased CD8 IFNγ+ tumor infiltrating T cells. These hu-CB-BRGS mice provide a relevant preclinical animal model to facilitate prioritization of hypothesis-driven combination immunotherapies.

Resistance exercise and lipoproteins in postmenopausal women.

The specific aims of this study were to quantify the effects of 12 weeks of resistance training, as well as a single session of resistance exercise on lipids and lipoproteins in obese, postmenopausal women. 21 obese, postmenopausal women, not on hormone replacement therapy (age=65.9 ± 0.5 yr; BMI=32.7 ± 0.8 kg/m(2)), were randomly assigned to control (n=12) and exercise (n=9) groups matched for age and BMI. For 12 weeks, 3 days/week, the exercise group performed 10 whole body resistance exercises (3 sets at 8-RM). Fasting (10 h) blood samples were collected immediately prior to and 24 h after the first and last exercise and control session. Serum was assayed for concentrations of total cholesterol, triglycerides, LDL-C, HDL-C, HDL 2-C, HDL 3-C, non-HDL-C and TC:HDL and LDL:HDL ratios. The exercise group exhibited a significant (P<0.01) improvement in muscular strength, but no change in BMI, body mass or body composition post-training. Total cholesterol, LDL-C and non-HDL-C were significantly (P<0.05) lower in the exercise compared to the control group following the 12 weeks of resistance training. Whole body resistance training provides obese, postmenopausal women a non-pharmacological approach for the reduction of lipid and lipoprotein-cholesterol concentrations.

Intramitochondrial crystalline inclusions in nonalcoholic steatohepatitis.

UNLABELLED: Mitochondrial dysfunction is an important element in the pathogenesis of nonalcoholic steatohepatitis (NASH). Intramitochondrial crystals (IMCs) are a well-documented morphological abnormality seen on transmission electron microscopy in this disease. It has been suggested that IMCs consist of phospholipids, but their exact composition remain uncertain many years after their discovery. Micellar phase transitions of phospholipid bilayers is a well-known but little-studied phenomenon in living systems. Its presence in the mitochondria of NASH would offer significant insight into the disease with possible therapeutic implications. We postulated that intramitochondrial disturbances in NASH are sufficient to produce such transitions and that their detection in fresh biopsies would therefore be a dynamic process. To test this, we performed a blinded, prospective analysis of fresh liver biopsy samples immediately fixed under different conditions. Quantitative transmission electron microscopy morphometry, performed by systematically counting total mitochondria and IMCs within areas of uniform dimension, showed a stepwise decline in IMCs with cooler fixation temperature in each subject studied. Randomization testing (Monte Carlo resampling) confirmed that the detection of IMCs was strongly dependent on fixation temperature (P < 0.0001). CONCLUSION: These results indicate that the intramitochondrial crystals characteristic of NASH are highly dynamic and unstable structures. The findings offer the strongest support yet for their origin in micellar phase transitions. We speculate that such transitions result from microenvironmental changes within the mitochondria and carry therapeutic implications, especially in regard to dietary manipulations of mitochondrial lipid composition.

Acute respiratory distress syndrome in a welder exposed to metal fumes.

A 43-year-old man began having malaise, chills, and fever 12 hours after cutting a galvanized steel grating with an acetylene torch at work. Over the next 72 hours, his symptoms persisted and became worse with progressive shortness of breath. He was admitted to the hospital and begun on antibiotics and steroids. The next day his condition had deteriorated to the point that he had to be intubated. Chest x-ray film and computed tomography showed patchy and interstitial infiltration bilaterally, consistent with acute respiratory distress syndrome. Open lung biopsy showed focal mild interstitial pneumonia. Multiple laboratory studies were negative for an infectious or an immune process. The patient remained on mechanical ventilation for 10 days and was discharged from the hospital 2 days after extubation. He continued to improve, with minimal symptoms and a return to normal activity levels several months after the incident with no continued treatment. Re-creation of his exposure was done under controlled circumstances, with air sampling revealing elevated air levels for cadmium and zinc and borderline levels of arsenic, manganese, lead, and iron.

Heart rate and blood pressure in panic disorder, major depression, and comorbid panic disorder with major depression.

Secondary major depression is common in patients with panic disorder (PD) and has been reported to worsen prognosis. Little is known about the effect of comorbid depression on the autonomic symptoms associated with PD. In this study, the heart rate and blood pressure of 44 patients with PD, 20 patients with major depression (MD), and 12 patients with both panic disorder and depression (PDD) were measured during postural challenge. Patients with PDD were found to have higher diastolic and systolic pressures throughout (P < 0.05), and to have a higher resting cardiac load (P < 0.05). This heightened autonomic arousal remained when the effects of age, race, and the frequency of panic attacks were removed. The results suggest that PDD patients may have increased autonomic dysfunction.

Mixed symptoms and syndromes of anxiety and depression: diagnostic, prognostic, and etiologic issues.

Patients with comorbid symptoms of anxiety and depression are common in clinical practice, and yet they continue to pose a diagnostic and therapeutic challenge to clinicians and researchers alike. This article is intended as a comprehensive review of current knowledge derived from a vast body of clinical research regarding the diagnostic interrelationships of anxiety and depressive disorders at both syndromal and subsyndromal levels. The prognostic implications of comorbidity are also reviewed. Biological linkages between the two disorders are explored, and recent biological theories attempting to explain and integrate the two disorders are presented as well. The article concludes with suggestions for further research.

Explained and unexplained medical symptoms in generalized anxiety and panic disorder: relationship to the somatoform disorders.

We have examined the numbers and types of symptoms in a sample of 90 patients with generalized anxiety disorder (GAD) and 77 patients with panic disorder (PD) collected from six different sites during the conduct of a multicenter clinical trial. This information was obtained utilizing the Health Questionnaire, a 47-item self-report list of medical symptoms, patterned after the Somatization Disorder section of the Diagnostic Interview Schedule. Although the patients in this sample had a wide variety of medically explained and unexplained physical symptoms, none of them qualified for a diagnosis of somatization disorder by DSM-III-R criteria. GAD and PD patients reported remarkably similar numbers of explained and unexplained medical symptoms. The panoply of somatic symptoms presented by these patients presents a formidable diagnostic challenge for clinicians. These findings suggest that the pattern of overutilization of medical services that is well documented for PD patients may also be found for GAD patients.

Serum cholesterol levels in patients with generalized anxiety disorder (GAD) and with GAD and comorbid major depression.

OBJECTIVE: To investigate risk for cardiovascular disease in patients with GAD, as well as the effects of comorbid major depression (MD). METHOD: Predrug-trial serum cholesterol and triglyceride levels were assessed in 38 patients with pure GAD and compared with those of 21 patients with mixed GAD and comorbid (MD). RESULTS: Significantly higher cholesterol and triglyceride levels were found in the GAD group. CONCLUSION: Increased noradrenergic activity may be responsible for elevations in lipid levels in patients with pure GAD.

The production of an active protein kinase C-delta in insect cells is greatly enhanced by the use of the basic protein promoter.

Occasionally, only a small percentage of recombinant proteins produced in the baculovirus expression system are functionally active. We had previously shown that the majority of protein kinase C-delta (PKC-delta) produced in insect cells was inactive; less than 1% of the expressed enzyme had lipid-dependent kinase activity. In this report, we have attempted to optimize the production of a catalytically active PKC-delta. Under optimum conditions, we were able to increase the levels of PKC-delta from 10-20% to about 65% of the total cellular protein; however, there was no increase in the levels of catalytically active enzyme. Expression of PKC-delta as a fusion protein or as a secreted protein also met with limited success. Under all conditions, expression of PKC-delta proteins under control of the strong polyhedrin promoter resulted in the production of large amounts of inactive enzyme. Expression under the control of the basic protein promoter, Pcor, resulted in the reduction of the levels of recombinant protein by a factor of about four, but the PKC-delta enzyme produced under these conditions was 10- to 15-fold more active. Thus, the earlier temporal expression of PKC-delta in insect cells resulted in the production of more active enzyme.

The cDNA sequence encoding human protein kinase C-zeta.

A 1779-bp complementary DNA (cDNA) that encodes protein kinase C-zeta (PKC-zeta) has been isolated from a human frontal cortex library using traditional plaque-screening methods and PCR screening. The deduced 592-amino-acid sequence of the human PKC-zeta clone has a 95-96% identity to those deduced from the previously described rat and mouse PKC-zeta clones.

Memory, benzodiazepines, and anxiety: integration of theoretical and clinical perspectives.

The benzodiazepines have been reported to cause anterograde amnestic effects. These impairments are likely mediated through the GABAA receptor complex, which contains the benzodiazepine receptor binding site. This GABAA receptor site is also believed to be the locus of the anxiolytic effects of the drugs as well, and recent research findings suggest that the anxiolytic and amnestic properties of the benzodiazepines may be functionally linked. Normal human memory function is extremely complex and requires the smooth integration of multiple aspects of cognition. On the basis of neuropsychological testing, the processes of memory have been divided into a variety of functions or stores that are interrelated, yet distinct. The amnestic effects of the benzodiazepines have been extensively studied within the context of this conceptual framework. Studies to date have shown that the memory deficits produced by the drugs are relatively selective and are primarily caused by impairments in the acquisition of newly learned information into long-term episodic storage. Benzodiazepines may impair other aspects of memory as well, but findings across studies have been inconsistent. The drugs clearly do not impair the recall of previously learned information, unlike anticholinergic drugs with amnestic effects, such as scopolamine. The amnestic effects of the benzodiazepines are dose-dependent, and can be predicted by pharmacokinetic/pharmacodynamic variables. In addition, multiple clinical factors may influence the likelihood of memory impairments. The amnestic effects of alprazolam are consistent with the drug effects of the benzodiazepines as a group.

Generalized anxiety disorder patients seek evaluation for cardiological symptoms at the same frequency as patients with panic disorder.

Although panic disorder (PD) and generalized anxiety disorder (GAD) have similar somatic symptoms, panic attacks with chest pain and/or palpitations may seem more likely to be mistaken for heart attacks because of their acute onset. One would therefore expect that PD patients are more likely than GAD patients to seek cardiological consultations. In a survey of 146 PD and 154 GAD patients entering a multi-site drug trial, we found virtually identical rates of such consults. Approximately 50% of each patient group sought medical evaluation for cardiac symptoms. Furthermore, 40% of each group had standard treadmill evaluations and 33% reported having an echocardiogram. This study suggests that future epidemiological studies in cardiology populations should include probes for generalized anxiety disorder.

Alprazolam as a neuroleptic adjunct in the emergency treatment of schizophrenia.

OBJECTIVE: While neuroleptics remain the mainstay of drug intervention in the emergency management of psychosis, a variety of agents have received study as alternatives or adjuncts to these drugs in an attempt to improve the safety and efficacy of acute treatment. The purposes of this study were to investigate the efficacy and safety of alprazolam as a neuroleptic adjunct for schizophrenic patients in psychotic relapse and to clarify the effects of combination treatment on specific aspects of the psychotic process. METHOD: Twenty-eight acutely psychotic patients with schizophrenia who were admitted to an emergency psychiatric service were randomly assigned to treatment with either haloperidol and alprazolam or haloperidol with placebo under double-blind conditions. Drug administration lasted 72 hours. RESULTS: Both groups improved significantly. The combination-treated group required significantly less medication and had 56% fewer dystonic reactions. The addition of alprazolam was most effective for symptoms of excitement and uncooperativeness, particularly in the initial hours of treatment. CONCLUSIONS: The combination of alprazolam and haloperidol seems to be the most effective for agitated patients, particularly in the first 48 hours of treatment. It may also result in fewer dystonic reactions.

Differential effects of alprazolam and buspirone upon acquisition, retention, and retrieval processes in memory.

A group of 125 healthy young adults was given a memory battery 1 hour after administration of alprazolam (0.5 and 1.0 mg), buspirone (5 and 10 mg), and placebo under single-dose, double-blind conditions. On the wordlist recall battery and a 24-hour retest, alprazolam appeared to impair acquisition and retention in dosages of 1.0 mg; buspirone did not adversely affect test performance at either dosage. On the basis of a 3-stage model of memory, a neuroantomical/neurophysiological model is proposed to explain the selective effects of the benzodiazepines on memory function.

A comparison of the single-dose effects of alprazolam, buspirone, and placebo upon memory function.

The effects of alprazolam and buspirone at varying dosages were compared with placebo in a battery of memory and cognitive tests in a sample of 125 young, healthy medical students by utilizing a single-dose, double-blind design. Although the dosages in this study did not impair visuomotor reaction time in any of the experimental groups, 1.0 mg of alprazolam did result in significantly impaired performance on selective aspects of memory function as assessed by the memory battery.

Toward a new alliance : psychiatric residents and family support groups.

In an attempt to assess the status of residency training in psychoeducational approaches for families of chronically ill patients, the authors conducted a national survey of U.S. residency training programs. Responses from 154 programs (75%) indicated a wide variation in time allotted, activities, and participants in such training among the respondents. Less than 50% of the programs indicated formal involvement of family support groups such as the National Alliance for the Mentally Ill (NAMI) and the National Depressive and Manie-Depressive Association (NDMDA). The authors describe a program they have initiated that incorporates organized family support groups in such training efforts.

Alcohol and substance abuse among schizophrenic patients presenting to an emergency psychiatric service.

To obtain information regarding patterns of alcohol and substance use, portions of the Diagnostic Interview Schedule (DIS) were administered to 53 schizophrenic patients who presented to the crisis service of an emergency room in a major general hospital. The number of individuals in the sample qualifying for a lifetime diagnosis of an alcohol abuse-related disorder was quite high (47%), and there was a strong correlation between disorders of alcohol abuse and the use of other drugs. Other than alcohol, marijuana was the most frequently abused drug. Individuals who qualified for a diagnosis of an alcohol-related disorder were compared with those who did not on a variety of diagnostic and demographic variables. The authors conclude by suggesting that the high-risk rates of drug use-related disorders reported in this sample may be due to the preferential use of emergency services by schizophrenic patients with alcohol and drug abuse-related disorders.