Opening caregiver minds: National Alliance for the Mentally Ill's (NAMI) provider education program.

The belief that poor parenting and dysfunctional families give rise to mental illness has been perpetuated by psychodynamic and family systems theories that lack supporting scientific evidence, and interventions based on these theories have failed to produce clinical improvements. Nevertheless the National Alliance for the Mentally III (NAMI) found that many clinical training programs continue to teach these outdated theories and interventions and that the mental health system is often destructive to family systems. This article describes a new 10-week program that is designed to educate service providers that will include families in the care of their chronically ill loved one. The program is based on a competence and adaptation rather than a pathology foundation and it shifts the discourse from causes to effects of illness.

The family experience of deinstitutionalization: insights from the closing of Central State Hospital.

Since the early 1970s, policy makers and researchers have expressed concern about the potential negative consequences of deinstitutionalization on families. This article summarizes results of a survey of family and lay caregivers of patients discharged from Central State Hospital, which closed in June 1994. The survey was designed to assess the impact of the closing on family members, including their attitudes, caregiving responsibilities, and involvement in the treatment of the patients. Results indicate that family members have mixed feelings about the closure. Family caregivers also reported that they have not been asked to take on significant amounts of the caregiving responsibilities since the clients were moved from the hospital. Family members described a significant reduction in the frequency of contact they had with their loved ones and with professional caregivers since the closure. Implications for behavioral health policy makers considering or planning closing or downsizing long-term care facilities are discussed.

Educational needs of families of mentally ill adults.

A total of 197 family members of mentally ill adults in Indiana responded to a survey about their preferences for family psychoeducation programs, including type of information, format, presenter, frequency and length of educational programs, setting, and cost. The findings of the survey, which was sent only to persons who were not members of the National Alliance for the Mentally Ill, indicated that family members throughout the state have consistent and persistent needs and clear preferences about educational programs. Of 11 educational topics listed, family members expressed the least interest in learning about their relative's substance abuse. They were ambivalent about whether to include patients in family education programs, but they clearly supported patient education.

Unique pancreatic carcinoma cDNA found via subtracted libraries with limited normal tissue and tumor from the same patient.

We have approached the problem of isolating clones unique to specific malignancies such as T-cell acute lymphoblastic leukemia (ALL) and pancreatic adenocarcinoma by using subtractive hybridization techniques. Our initial studies involved using normal donor tissue (i.e., normal blood donors for ALL and cadaver renal transplant donors for normal pancreatic tissue) and cultured malignant cell lines. It occurred to us that normal pancreatic tissue from the same patient source as that of the malignant tissue might subtract out normal sequences more readily and enrich clones unique to pancreatic adenocarcinoma because of patient/donor identity. Using such a method meant that the amounts of tissue for overcoming this obstacle. We constructed independent UNI-ZAP-XR cDNA libraries (normal and malignant) and used them to amplify either the normal or malignant cDNA prior to subtractive hybridization. We then obtained rescued single stranded cDNA from the malignant ZAP library. The RNA which was not hybridized was isolated. The process was repeated and a double subtraction was effected. The residual non-hybridized RNA was used as a template for first and second strand synthesis. After the EcoRI adaptors were ligated to the double stranded cDNA it was cloned into Lambda ZAP II arms to form a double subtracted malignant cDNA library. A subtracted probe was prepared from the double subtracted cDNA library. Single stranded cDNA was rescued, double stranded plasmid was made, the plasmid DNA was digested with EcoRI, the digested DNA was run on a 1% SeaPlaque gel, and the insert cDNA was recovered using Ultra-Free MC and Ultra-Free Probind filters. The subtracted malignant cDNA library was probed with the subtracted probe and with normal cDNA (obtained from the normal ZAP library) and those plaques which were positive per the subtracted probe and negative per the normal cDNA were isolated; their cDNA inserts are being further characterized.

The role of activation of neutrophils and microvascular pressure in acute pulmonary edema.

Activated polymorphonuclear neutrophils (PMN) can mediate vascular injury in the lung. This study compared activated aggregate PMN (emboli) to activated PMN that were previously adhered to the microvasculature (non-embolic) in the isolated perfused rat lung. Permeability and microvascular pressure (Pmv), components of PMN-induced edema, were examined by continuous measurement of wet weight, pulmonary arterial and left atrial pressures, and by intermittent determination of double occlusion pressure. PMN that were activated with phorbol myristate acetate and then perfused into the lung formed aggregates that lodged primarily in the precapillary bed, increasing arterial resistance. Although these PMN had minimal direct contact with the capillary endothelium, edema rapidly developed and Pmv was progressively elevated. If PMN were allowed to adhere in the capillary bed, a minimal and nonprogressive increase in Pmv and lung weight occurred. When these adherent PMN were then activated, there was a progressive rise in both Pmv and lung weight. The free radical scavenger catalase prevented this edema formation but not the rise in pressure. In control lungs with matched elevation of Pmv, edema did not develop. In another group of lungs with activation of pre-adherent PMN in which Pmv was maintained at control levels, edema formation was greatly delayed. These data show that: (1) the activated PMN free radical products alone caused permeability injury in the lung because neither contact of the PMN with the capillary endothelium nor embolization was necessary, and (2) increased Pmv does not cause edema but greatly increases the rate of edema formation when the endothelium is injured.

Comparison of in vivo effects of intravenous infusion of N-formyl-methionyl-leucyl-phenylalanine and phorbol myristate acetate in rabbits.

Comparison of the physiologic responses in rabbits to the intravenous infusion of two polymorphonuclear neutrophil (PMN) activators, N-formyl-methionyl-leucyl-phenylalanine (FMLP) and phorbol myristate acetate (PMA), has revealed marked differences in kinetics for activation between these agents. FMLP infusion was associated with maximally increased respiratory rates (RR), a maximally decreased mean blood pressure (MBP), and a maximally decreased absolute granulocyte count (AGC), all within the first 5 min after infusion. However, there were no significant differences between RR, MBP, and AGC of FMLP-treated animals and controls, 15 min postinfusion and after. On the other hand, PMA did not cause significant changes in RR or MBP until 30 min and 2 h postinfusion, respectively. Previous work has demonstrated that both FMLP and PMA stimulate the PMN metabolically in vitro via the same respiratory burst enzyme, NADPH oxidase, but that each of these activators demonstrates kinetics which are different from the other. Thus, these data from an in vivo study support previous in vitro findings and offer further evidence that the neutropenia and cardiopulmonary alterations following intravenous infusion of FMLP and PMA may be caused by metabolic activation of the blood PMN.

Effect of vitamin E on FMLP-induced activation of rabbit polymorphonuclear leukocytes.

Granulocytes of vitamin E-treated rabbits were compared to granulocytes from placebo-treated rabbits. Granulocytes were isolated from rabbit peripheral blood by a new method employing Percoll and gelatin sedimentation. Vitamin E-treated cells showed less adherence to rabbit aortic endothelium when stimulated with FMLP. FMLP receptor numbers and affinity were not significantly different. Resting cell surface and baseline transmembrane potential were similar in both cell types. Decrease in cell surface potential with FMLP was comparable in vitamin E- and placebo-treated cells. Vitamin E-treated PMN depolarized more and hyperpolarized more rapidly than placebo cells. Thus vitamin E-treated PMNs show differences in the early events of PMN activation. These may contribute to the lower stimulated adherence observed with vitamin E-treated cells.

In vitro and in vivo effects of treatment by platelet-activating factor on N-formyl-met-leu-phe-mediated responses of polymorphonuclear leucocytes.

Two chemoattractants, the peptide N-formyl-met-leu-phe (FMLP), and the ether phospholipid, platelet activating factor (PAF), each stimulate a variety of in vitro responses in polymorphonuclear leucocytes (PMN). Because often more than one inflammatory mediator is active during inflammation, we determined the effect on PMN of sequential stimulation with these two agents. Before FMLP stimulation, human PMN were exposed to PAF, at concentrations which gave little or no response when administered alone. PAF enhanced FMLP-elicited superoxide release in a dose-dependent fashion. Likewise, release of granular lysozyme from the cells was increased in PAF treated cells. Similar treatment with other phospholipids, including the lyso derivation of PAF, failed to produced these effects. Incubation with nordihydroguaiaretic acid, an inhibitor of arachidonic acid metabolism, had little effect on the enhancement of lysozyme release by PAF. To determine if enhancing effects by PAF might occur also in vivo, we studied rabbits receiving PAF and/or FMLP intravenously. When rabbits received 0.01 micrograms PAF (a dose which does not elicit the sustained neutropenia observed with higher doses of PAF) followed by 0.05 micrograms FMLP the absolute granulocyte count (AGC) dropped at 1 min (46 +/- 11% of original value), and continued to fall (24 +/- 12% at 10 min). Controls, treated with the suspending fluid for PAF, and then 0.05 micrograms FMLP, had a similar 1 min AGC value, but at 10 min AGC returned to 65 +/- 6.1% (P less than 0.001 for comparison of 10 min values). Thus PAF pretreatment enhanced FMLP-elicited granulocytopenia in vivo. Study of in vitro human PMN aggregation revealed that, at certain relative concentrations of PAF and FMLP, aggregation was enhanced. These studies show that both in vitro and in vivo responses of FMLP-stimulated PMN may be exaggerated by pre-exposure to PAF.

Amniotic basement membrane: a barrier to neutrophil invasion.

Polymorphonuclear neutrophils activated by N-formylmethionyl-leucylphenylalanine are able to cross the human amnion as shown by Russo et al. (1981). With the same technique, we added polymorphonuclear neutrophils to either the epithelial or stromal surface of the amnion. The lower compartment of the incubation chamber contained 10(-8) mol/L of N-formylmethionyl-leucylphenylalanine. Of 24 amnion tested, only one showed polymorphonuclear neutrophil permeation. Ultrastructural observations indicated that the basal lamina and the zona reticularis of the basement membrane acted as barriers to polymorphonuclear neutrophil invasion. Recent studies indicate that the basal lamina and the zona reticularis of the basement membrane contain collagen type V. No data support the existence of collagenase of polymorphonuclear neutrophil origin that cleaves collagen type V. If enzymatic degradation of extracellular matrix components play a role in polymorphonuclear neutrophil invasion, it is possible that type V collagen acts as a barrier to polymorphonuclear neutrophil invasion in the human amnion.

Vitamin E attenuates the effects of FMLP on rabbit circulating granulocytes.

Exposure of circulating rabbit granulocytes to the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (FMLP) in vivo results in transient granulocytopenia, hypotension, and cardiorespiratory distress. The effectiveness of vitamin E in attenuating these responses was tested. Vitamin E accelerated the rate of return of granulocytes to the peripheral circulation after FMLP-induced granulocytopenia and mitigated the hypotension. The reversible adherence of FMLP-stimulated granulocytes to endothelium offers a plausible mechanism to explain the transient granulocytopenia. From in vitro studies it was found that FMLP-activated granulocytes from animals treated with vitamin E showed decreased adherence to cultivated aortic endothelial monolayers when compared with FMLP-activated granulocytes from control animals.

Plasma lactoferrin reflects granulocyte activation in vivo.

N-formyl-met-leu-phe (FMLP) causes polymorphonuclear leukocytes (PMN) to secrete and become "sticky" in vitro. We related these events to in vivo FMLP-induced neutropenia. FMLP was intravenously administered to anesthetized rabbits in doses ranging from 0.01 microgram to 1.0 microgram. Controls received phosphate-buffered saline (PBS), the diluent for FMLP. Blood pressure, respiratory rate, and arterial Po2 were monitored. High and intermediate doses of FMLP caused a dramatic but transient decrease in blood pressure and an increase in respiratory rate. Prior to FMLP infusion, plasma lactoferrin level was 6.4 +/- 4.1 micrograms/ml, and the absolute granulocyte account (AGC) was 2008 +/- 1229 (mean +/- SD). There was a positive linear correlation between AGC and plasma lactoferrin level prior to injection of FMLP (R2 = 0.74, p less than 0.01). At 1 min after FMLP injection, the percent change in AGC decreased as an exponential function of dose to as low as 10% of baseline (R2 = 0.86, p = 0.002) and plasma concentration of lactoferrin increased as an exponential function of dose to as high as 30 micrograms/ml (R2 = 0.84, p = 0.006). Thus, FMLP-induced neutropenia is associated with increased levels of plasma lactoferrin, suggesting that PMN are induced to degranulate in vivo.

The effect of vitamin E on rabbit neutrophil activation.

Chemoattractants such as N-formylmethionyl leucyl phenylalanine (FMLP) cause neutropenia in vivo. The sequestered neutrophils may block the microvasculature and contribute to respiratory distress. Neutrophils from humans receiving 1600 units vitamin E per day have reduced oxidative activity. To test whether vitamin E attenuates the responses of neutrophils to FMLP in vivo we gave rabbits four daily intramuscular injections of 100 mg vitamin E. Serum levels of the vitamin were 2.34 +/- 0.15 mg% compared to 0.19 +/- 0.04 mg% in control rabbits receiving placebo injections. On the fifth day testing was done before and after injecting FMLP. Variables monitored were the absolute granulocyte count (AGC), systolic, diastolic and mean blood pressures (MBP), heart rate, PO2, PCO2, pH and respiratory rate. When 0.5 microgram FMLP was injected intravenously the AGC decreased (at 2.5 min the percentage change was -89.7 +/- 8.0 with vitamin E and -97.0 +/- 2.7 without vitamin E; P = 0.2). MBP decreased also (% change, -29.0 +/- 13.0 with vitamin E and -36.3 +/- 16.0, without vitamin E). By 15 min recovery was seen (AGC % change, -26.0 +/- 17 with vitamin E and -78.7 +/- 10.5, without vitamin E; P = 0.01; MBP % change, -9.3 +/- 3.8 with vitamin E and -52.3 +/- 10.1 without vitamin E). Chromatographic analysis of serum extracts revealed increases in 6-keto-PGF1 alpha after stimulation. Studies with [3h]thymidine-labelled neutrophils showed that the sequestered cells return to the circulation. Vitamin E might facilitate this return by altering the adherence of neutrophils to endothelium. This possibility was tested by measuring the adherence to cultivated rabbit aorta endothelial monolayers of FMLP-stimulated neutrophils from vitamin E-treated rabbits. The percentage of neutrophils adhering was 32.5 +/- 3.5 with vitamin E and 60.0 +/- 7.1, without vitamin E. Thus vitamin E promotes the return of neutrophils to the circulation after chemotactic challenge and may do so by reducing their adherence to endothelium.