Prevalence of psychosocial issues among pregnant women who do and do not use illicit substances.

OBJECTIVE: The American College of Obstetricians and Gynecologists (ACOG) recommends providers screen their prenatal patients for 11 psychosocial issues because they affect patient mental and physical well-being. The prevalence and co-occurrence of these issues have rarely been compared among pregnant women who do and do not report recent illicit substance use. METHOD: Seven psychosocial issues identified by ACOG were operationalized using National Survey on Drug Use and Health variables. We report weighted prevalence and adjusted risk ratios (ARR) for these issues in pregnant women who did versus did not report past-month illicit substance use. RESULTS: Pregnant women (n = 3,657) who reported past-month illicit substance use (6.3%; 95% CI [5.4-7.3]) had significantly higher prevalence of almost all psychosocial issues examined, including past-month cigarette smoking (44.9% versus 9.5%; ARR = 2.84, 95% CI [2.21-3.65]); past-month alcohol use, 36.1% versus 7.9%; ARR = 4.71 (3.59-6.18); serious past-month distress, 23.0% versus 5.0%; ARR = 3.51 (2.39-5.15); no health insurance, 11.7% versus 6.2%; ARR = 1.71 (1.07-2.74); and receipt of food stamps, 45.0% versus 24.0%; ARR = 1.40 (1.18-1.67). Moving 3 + times in the past year followed a similar pattern, but results were compatible with there being no difference, 10.6% versus 5.5%; ARR = 1.39 (0.86-2.25). The majority of pregnant women reporting illicit substance use endorsed experiencing ≥ 2 psychosocial issues while the majority of those who did not report illicit substance use did not endorse any. CONCLUSIONS: Pregnant women who use illicit substances experience higher prevalence and greater co-occurrence of psychosocial issues compared to those who do not, reinforcing recommendations for multidisciplinary approaches to care. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Prevalence of Psychosocial Issues Among Pregnant Women Who Do and Do Not Use Illicit Substances.

Objective It is often believed that pregnant women who use illicit substances are more likely to experience psychosocial issues like smoking, depression, and inadequate health care compared to pregnant women who do not. However, the prevalence of these psychosocial issues has rarely been calculated and compared using nationally representative data. Methods Important psychosocial issues identified by the American College of Obstetricians and Gynecologists were operationalized using variables in the National Survey on Drug Use and Health. We report weighted prevalence and age-adjusted odds ratios for these issues in pregnant women who did vs. did not report past-month illicit substance use. Results Pregnant women (n = 3,657) who reported past-month illicit substance use (6.3%; 95% CI 5.4-7.0) had significantly higher rates of almost all psychosocial issues examined, including past-month cigarette smoking (44.9% vs. 6.5%; age-adjusted odds ratio (AOR) = 7.14 (95% CI 4.98-10.20)); past-month alcohol use (36.1% vs. 7.8%; AOR = 6.80 (4.69, 9.86)); serious past-month distress (23.0% vs. 5.0%; AOR = 4.99 (3.07-8.11)); no health insurance (11.7% vs. 6.2%; AOR = 1.79 (1.07-2.99)); and receipt of food stamps (45.0% vs. 24.0%; AOR = 2.26 (1.55-3.29)). Moving 3 + times in the past year followed a similar pattern, but results were compatible with there being no difference between groups (10.6% vs. 5.5%; AOR = 1.59 (0.95-2.66)). In contrast to other issues examined, English language proficiency was higher among those who reported illicit substance use (4.7% vs. 0.4%; AOR = 0.08 (0.01-0.63)). Conclusions Pregnant women who use illicit substances experience higher rates of most psychosocial issues compared to those who do not, reinforcing recommendations for multidisciplinary approaches to care.

Preliminary side effect profile, safety, and tolerability of hormonal contraception among women receiving opioid-agonist therapy.

OBJECTIVE(S): To characterize for the first time the side effect profile, safety, and tolerability of hormonal contraception among women receiving opioid-agonist therapy. STUDY DESIGN: We conducted a secondary analysis of data collected from participants in a three-arm randomized controlled trial (N = 138) aimed at increasing effective contraceptive use among women receiving opioid-agonist therapy. Participants in the 2 intervention conditions (n = 90) had free access to hormonal contraception at each of the 14 visits scheduled during the 6-month intervention. Contraceptive use and side effects were recorded at each visit; participants could change methods or discontinue use at any time. Verbatim side effects were classified using Medical Dictionary for Regulatory Activities (MedDRA) terminology. RESULTS: Of 67 participants reporting hormonal contraceptive use, 29 (43%) initiated implants, 14 (21%) intrauterine devices, 13 (19%) combined pills, 11 (16%) progestin-only pills, 10 (15%) injectables, 1 (2%) ring, and 1 (2%) patch; the average (±standard deviation) duration of use was 129 ± 55, 129 ± 60, 108 ± 62, 102 ± 61, 111 ± 31, 145, and 18 days, respectively. A total of 321 side effects were reported by 55 (82%) participants. Fifty (75%) participants reported menstrual cycle changes/uterine bleeding, followed by headaches (16, 24%), weight gain (15, 22%), and abdominal pain or nausea/vomiting (11, 16%). No serious side effects were reported. Twelve participants (18%) changed methods and 13 (19%) discontinued all hormonal contraceptive use. CONCLUSION(S): The hormonal contraceptive side effects reported by this small group of women receiving opioid-agonist therapy appear consistent with those reported by the general population, was generally well-tolerated, and did not raise safety concerns. IMPLICATIONS: These results provide important preliminary evidence that hormonal contraceptive use produces a familiar side effect profile and is well-tolerated by women receiving opioid-agonist therapy, although studies with larger samples followed over longer periods of time with appropriate comparison conditions are needed to fully assess tolerability and safety.

Financial Analysis of Cardiac Rehabilitation and the Impact of COVID-19.

PURPOSE: Provision of phase 2 cardiac rehabilitation (CR) has been directly impacted by coronavirus disease-19 (COVID-19). Economic analyses to date have not identified the financial implications of pandemic-related changes to CR. The aim of this study was to compare the costs and reimbursements of CR between two periods: (1) pre-COVID-19 and (2) during the COVID-19 pandemic. METHODS: Health care costs of providing CR were calculated using a microcosting approach. Unit costs of CR were based on staff time, consumables, and overhead costs. Reimbursement rates were derived from commercial and public health insurance. The mean cost and reimbursement/participant were calculated. Staff and participant COVID-19 infections were also examined. RESULTS: The mean number of CR participants enrolled/mo declined during the pandemic (-10%; 33.8 ± 2.0 vs 30.5 ± 3.2, P = .39), the mean cost/participant increased marginally (+13%; $2897 ± $131 vs $3265 ± $149, P = .09), and the mean reimbursement/participant decreased slightly (-4%; $2959 ± $224 vs $2844 ± $181, P = .70). However, these differences did not reach statistical significance. The pre-COVID mean operating surplus/participant ($62 ± $140) eroded into a deficit of -$421 ± $170/participant during the pandemic. No known COVID-19 infections occurred among the 183 participants and 14 on-site staff members during the pandemic period. CONCLUSIONS: COVID-19-related safety protocols required CR programs to modify service delivery. Results demonstrate that it was possible to safely maintain this critically important service; however, CR program costs exceeded revenues. The challenge going forward is to optimize CR service delivery to increase participation and achieve financial solvency.

Efficacy and Cost-Benefit of Onsite Contraceptive Services With and Without Incentives Among Women With Opioid Use Disorder at High Risk for Unintended Pregnancy: A Randomized Clinical Trial.

Importance: Rates of in utero opioid exposure continue to increase in the US. Nearly all of these pregnancies are unintended but there has been little intervention research addressing this growing and costly public health problem. Objective: To test the efficacy and cost-benefit of onsite contraceptive services with and without incentives to increase prescription contraceptive use among women with opioid use disorder (OUD) at high risk for unintended pregnancy compared with usual care. Design, Setting, and Participants: A randomized clinical trial of 138 women ages 20 to 44 years receiving medication for OUD who were at high risk for an unintended pregnancy at trial enrollment between May 2015 and September 2018. The final assessment was completed in September 2019. Data were analyzed from October 2019 to March 2021. Participants received contraceptive services at a clinic colocated with an opioid treatment program. Interventions: Participants were randomly assigned to receive 1 of 3 conditions: (1) usual care (ie, information about contraceptive methods and community health care facilities) (n = 48); (2) onsite contraceptive services adapted from the World Health Organization including 6 months of follow-up visits to assess method satisfaction (n = 48); or (3) those same onsite contraceptive services plus financial incentives for attending follow-up visits (n = 42). Main Outcomes and Measures: Verified prescription contraceptive use at 6 months with a cost-benefit analysis conducted from a societal perspective. Results: In this randomized clinical trial of 138 women (median age, 31 years [range, 20-44 years]), graded increases in verified prescription contraceptive use were seen in participants assigned to usual care (10.4%; 95% CI, 3.5%-22.7%) vs contraceptive services (29.2%; 95% CI, 17.0%-44.1%) vs contraceptive services plus incentives (54.8%; 95% CI, 38.7%-70.2%) at the 6-month end-of-treatment assessment (P < .001 for all comparisons). Those effects were sustained at the 12-month final assessment (usual care: 6.3%; 95% CI, 1.3%-17.2%; contraceptive services: 25.0%; 95% CI, 13.6%-39.6%; and contraceptive services plus incentives: 42.9%; 95% CI, 27.7%-59.0%; P < .001) and were associated with graded reductions in unintended pregnancy rates across the 12-month trial (usual care: 22.2%; 95% CI, 11.2%-37.1%; contraceptive services: 16.7%; 95% CI, 7.0%-31.4%; contraceptive services plus incentives: 4.9%; 95% CI, 0.6%-15.5%; P = .03). Each dollar invested yielded an estimated $5.59 (95% CI, $2.73-$7.91) in societal cost-benefits for contraceptive services vs usual care, $6.14 (95% CI, $3.57-$7.08) for contraceptive services plus incentives vs usual care and $6.96 (95% CI, $0.62-$10.09) for combining incentives with contraceptive services vs contraceptive services alone. Conclusions and Relevance: In this randomized clinical trial, outcomes with both onsite contraceptive service interventions exceeded those with usual care, but the most efficacious, cost-beneficial outcomes were achieved by combining contraceptive services with incentives. Colocating contraceptive services with opioid treatment programs offers an innovative, cost-effective strategy for preventing unintended pregnancy. Trial Registration: ClinicalTrials.gov Identifier: NCT02411357.

Contraceptive Knowledge among Females and Males Receiving Medication Treatment for Opioid Use Disorder Compared to Those Seeking Primary Care.

BACKGROUND: Rates of unintended pregnancy among women receiving medication treatment for opioid use disorder (mOUD) are high, likely due in part to low rates of contraceptive use. Lack of knowledge about contraception may be contributing to inadequate contraceptive use. Objectives: To compare contraceptive knowledge among women and men receiving mOUD relative to a comparison group seeking primary care. Methods: We surveyed 332 reproductive-age women and men receiving mOUD or primary care with the recently validated 25-item Contraceptive Knowledge Assessment. We examined overall differences between patient groups and between females and males using two-way analyses of variance; individual item differences were tested using logistic regression. Results: The mean percent of total correct responses was lower among individuals receiving mOUD (n = 167) relative to the comparison group (n = 165), 47.7% vs. 53.8%, respectively (p < .001), and higher among females (n = 169) relative to males (n = 163), 56.1% vs. 45.2%, respectively (p < .001). Individual item analyses revealed patient group and sex differences primarily in the areas of contraceptive efficacy, attributes of contraceptive methods, and fertility awareness. Conclusion: While there were modest differences between patient groups and sexes, the results suggest a deficit in contraceptive knowledge across all groups. Substance use disorder treatment clinics could be an opportunistic setting in which to provide accurate information about contraception to patients with OUD and other substance use disorders.

Perceptions of long-acting reversible contraception among women receiving medication for opioid use disorder in Vermont.

OBJECTIVES: To evaluate perceptions of long-acting reversible contraceptives (LARC) among women receiving medication for opioid use disorder. STUDY DESIGN: Cross-sectional survey of 200 women receiving medication for opioid use disorder in Vermont. RESULTS: A considerable proportion of women receiving medication for opioid use disorder in Vermont reported previous use of an IUD (40%) and/or a subdermal contraceptive implant (16%); the majority of prior LARC users were satisfied with their IUD (68%) or their implant (74%). Of the 38% of participants who had never considered IUD use, 85% percent (64/75) said that they knew nothing or only a little about IUDs. Of the 61% of participants who had never considered an implant, 81% percent (98/121) said that they knew nothing or only a little about the contraceptive method. The most commonly reported reasons for a lack of interest in the IUD and/or implant were concerns about side effects and preference for a woman-controlled method. CONCLUSIONS: Gaps in LARC knowledge are common among those who have not used LARCs and concerns about side effects and preferences for a woman-controlled method limit some women's interest in these contraceptives. Additionally, reasons for dissatisfaction among past users are generally similar for IUD and implant and include irregular bleeding and having a bad experience with the method. IMPLICATIONS: Efforts to increase awareness of LARC methods among women receiving medication for opioid use disorder should address concerns about side effects and reproductive autonomy and encourage satisfied LARC users to share their experiences with their social networks.

Innovative approaches to reduce unintended pregnancy and improve access to contraception among women who use opioids.

Dramatic increases in the rate of opioid use disorder (OUD) during pregnancy have been paralleled by substantial increases in the number of neonates diagnosed with neonatal abstinence syndrome (NAS). Women with OUD have reliably reported high rates of unintended pregnancy and a number of studies also indicate they desire easier access to contraception. Recent statements from the Centers for Disease Control and Prevention and the American Academy of Pediatrics/American College of Obstetricians and Gynecologists have drawn increased attention to efforts to prevent unintended pregnancy and improve access to contraception among women with OUD. We briefly review a number of innovative clinical approaches in these areas, including efforts to integrate family planning services into substance use disorder (SUD) treatment and other settings that serve people with OUD and interventions that aim to make family planning a higher priority among women with OUD. Results suggest many of these approaches have led to increases in contraceptive use and may aid in efforts to reduce unintended pregnancy and improve access to contraception among women with OUD now and in the future.

Knowledge of and concerns about long-acting reversible contraception among women in medication-assisted treatment for opioid use disorder.

OBJECTIVES: To assess interest in, concerns about and knowledge of long-acting reversible contraceptives (LARC) among women in medication-assisted treatment (MAT) for opioid use disorder who were at risk for unintended pregnancy. STUDY DESIGN: Women in MAT completed a survey on contraceptive use, attitudes and knowledge, including LARC methods, as part of eligibility screening for an ongoing trial evaluating family planning interventions for this population. RESULTS: Eighty-three women at risk for unintended pregnancy completed the survey, and a subset of 51 completed supplemental questions about implants. All participants had heard of IUDs and 75/83 (90%) had heard of implants, but only 34/83 (41%) and 14/51 (27%) reported being likely to use IUDs and implants, respectively. Thirty-five women reported they were unlikely to use IUDs, with 29/35 (83%) citing unspecified "other reasons" for this position and 24/35 (69%) endorsing concerns about side effects. Seventeen women reported they were unlikely to use implants, with 8/17 (47%) citing "other reasons" and 9/17 (53%) and 10/17 (59%) reporting concerns about insertion and removal, respectively. Participants reported they knew "a little" about LARCs, but only their knowledge of implants was limited. Only 50/83 (60%) participants were aware of the superior effectiveness of IUDs and 26/51 (51%) were aware of the superior effectiveness of implants relative to other reversible methods. CONCLUSIONS: Participants reported relatively low interest in LARCs. Many women had unspecified reasons for not using LARCs. Participants also lacked information about LARC safety and effectiveness, especially with regard to implants. IMPLICATIONS: Women in medication-assisted treatment for opioid use disorder are at high risk of unintended pregnancy, yet contraceptive use is low and use of less effective methods is common. Women in MAT may benefit from efforts to increase knowledge about implants and assuage concerns about their insertion and removal as well as more general efforts to improve knowledge about the relative efficacy of LARCs. Many women in MAT endorsed unspecified "other reasons" for not using LARCs, which should be explored further.

Insights into the regulation of TNF-alpha production in human mononuclear cells: the effects of non-specific phosphodiesterase inhibition.

OBJECTIVE: The objective of this study was to determine the effect of nonspecific phosphodiesterase inhibition on transcription factor activation and tumor necrosis factor-alpha (TNF-alpha) production in lipopolysaccharide (LPS)-stimulated human mononuclear cells. INTRODUCTION: The production of TNF-alpha following LPS stimulation is one of the key steps in bacterial sepsis and inflammation. The mechanism by which phosphodiesterase inhibition alters TNF-alpha production in the presence of LPS remains unclear. METHODS: Human mononuclear cells were stimulated with LPS (1 microg/mL), in the presence and absence of Pentoxifylline (PTX; 20 mM), a nonspecific phosphodiesterase inhibitor. Western blotting of phosphorylated cytoplasmic I-kBalpha, nuclear factor-kB p65 (NF-kB), and nuclear cAMP-response element binding protein (CREB) was performed. DNA binding of NF-kB and CREB was verified by electrophoretic mobility shift assay. TNF-a levels were determined in the supernatant of stimulated cells in the presence and absence Protein kinase A inhibition by an enzyme-linked immunosorbent assay (ELISA). RESULTS: PTX was demonstrated to significantly reduce cytoplasmic I-kBalpha phosphorylation, nuclear p65 phosphorylation, and the DNA binding activity of NF-kB. In contrast, PTX markedly enhanced the phosphorylation and DNA binding activity of CREB. Cells concomitantly treated with PTX and LPS secreted similar levels of TNF-a in the presence and absence Protein kinase A inhibition. DISCUSSION: The increased level of cAMP that results from phosphodiesterase inhibition affects cytoplasmic and nuclear events, resulting in the attenuation of NF-kB and the activation of CREB transcriptional DNA binding through pathways that are partially Protein kinase A-independent. CONCLUSION: PTX-mediated phosphodiesterase inhibition occurs partially through a Protein kinase A-independent pathway and may serve as a useful tool in the attenuation of LPS-induced inflammation.

Insights into the regulation of TNF-a production in human mononuclear cells: the effects of non-specific phosphodiesterase inhibition

OBJECTIVE: The objective of this study was to determine the effect of nonspecific phosphodiesterase inhibition on transcription factor activation and tumor necrosis factor-alpha (TNF-a) production in lipopolysaccharide (LPS)-stimulated human mononuclear cells. INTRODUCTION: The production of TNF-a following LPS stimulation is one of the key steps in bacterial sepsis and inflammation. The mechanism by which phosphodiesterase inhibition alters TNF-a production in the presence of LPS remains unclear. METHODS: Human mononuclear cells were stimulated with LPS (1 µg/mL), in the presence and absence of Pentoxifylline (PTX; 20 mM), a nonspecific phosphodiesterase inhibitor. Western blotting of phosphorylated cytoplasmic I-kBa, nuclear factor-kB p65 (NF-kB), and nuclear cAMP-response element binding protein (CREB) was performed. DNA binding of NF-kB and CREB was verified by electrophoretic mobility shift assay. TNF-a levels were determined in the supernatant of stimulated cells in the presence and absence Protein kinase A inhibition by an enzyme-linked immunosorbent assay (ELISA). RESULTS: PTX was demonstrated to significantly reduce cytoplasmic I-kBa phosphorylation, nuclear p65 phosphorylation, and the DNA binding activity of NF-kB. In contrast, PTX markedly enhanced the phosphorylation and DNA binding activity of CREB. Cells concomitantly treated with PTX and LPS secreted similar levels of TNF-a in the presence and absence Protein kinase A inhibition. DISCUSSION: The increased level of cAMP that results from phosphodiesterase inhibition affects cytoplasmic and nuclear events, resulting in the attenuation of NF-kB and the activation of CREB transcriptional DNA binding through pathways that are partially Protein kinase A-independent. CONCLUSION: PTX-mediated phosphodiesterase inhibition occurs partially through a Protein kinase A-independent pathway and may serve as a useful tool in the attenuation of LPS-induced inflammation.

The effects of a novel resuscitation strategy combining pentoxifylline and hypertonic saline on neutrophil MAPK signaling.

BACKGROUND: The combination of hypertonic saline (HS) and pentoxifylline (PTX) has been shown to synergistically downregulate neutrophil oxidative burst in vitro. We investigated the effects of HS/PTX on human neutrophil mitogen-activated protein kinase (MAPK) signaling and the role of Protein kinase A (PKA) in this process. METHODS: Isolated neutrophils were treated with PTX (2 mmol/L), HS10 (10 mmol/L above isotonicity), and HS40 (40 mmol/L above isotonicity) alone or in combination for determination of intracellular cyclic adenosine monophosphate (cAMP) concentrations. Human neutrophils were stimulated with f-methionyl-leucyl-phenylalanine (fMLP) (1 micromol/L) before the treatments above in both the presence and the absence of PKA inhibition for Western blot analysis of MAPK p38 and extracellular signal-related kinase 1/2 (ERK 1/2) phosphorylation. RESULTS: Concomitant exposure to HS/PTX results in an additive increase in intracellular cAMP. fMLP-induced ERK 1/2 phosphorylation was synergistically attenuated by HS/PTX. Both PTX and HS reduced p38MAPK phosphorylation. No additive effect was observed with combined treatment. Although PKA inhibition abrogated the effects of PTX, HS retained some capacity to attenuate MAPK phosphorylation. CONCLUSION: HS/PTX is more effective in attenuating neutrophil ERK signaling than either component alone, whereas both components alone or in combination produced comparable results with p38MAPK. Although PTX functions primarily through PKA activation, HS may suppress neutrophils through a partially PKA-independent mechanism.

Pentoxifylline attenuates stored blood-induced inflammation: A new perspective on an old problem.

BACKGROUND: Blood transfusion is a risk factor for many inflammatory processes. Its supernatant fraction has been proven to activate neutrophils. We hypothesized that pentoxifylline (PTX) would attenuate stored blood-induced neutrophil activation and pro-inflammatory mediator production. METHODS: Whole blood was incubated with HBSS, LPS (100 microg/mL), leukoreduced PRBC supernatant + LPS, or supernatant + LPS + PTX (2 mmol/L). TNF-alpha levels were measured by ELISA. MMP-9 was evaluated with zymography. Neutrophil CD66b expression was determined by flow cytometry in blood treated with HBSS, fMLP (1 micromol/L), supernatant + fMLP, or supernatant + fMLP + PTX. RESULTS: TNF-alpha levels were elevated in both the LPS and supernatant + LPS groups (100%; P < 0.01 and 120%; P < 0.01, respectively). PTX administration resulted in a 106% decrease in TNF-alpha (P < 0.0001). MMP-9 levels were increased in all groups. Administration of PTX to the supernatant + LPS group generated a 33% decrease in MMP-9 levels, which was not statistically significant (P < 0.4). Upregulation of CD66b expression was seen in LPS and supernatant + LPS groups. Significant attenuation was seen with PTX (47%; P < 0.01). CONCLUSIONS: PTX downregulates CD66b and TNF-alpha expression in supernatant-induced whole blood. Because blood transfusion can contribute to inflammatory injury, the adjunctive use of PTX may have therapeutic potential.

Neutrophil degranulation and the effects of phosphodiesterase inhibition.

BACKGROUND: Neutrophils play a major role as the first line in host defense after exposure to bacterial products. However, an exaggerated inflammatory response characterized by overwhelming neutrophil activation can be injurious to the host. Pentoxifylline (PTX), a nonspecific phosphodiesterase inhibitor, has been shown to attenuate neutrophil oxidative burst and decrease proinflammatory mediator synthesis. We hypothesized that PTX down-regulates neutrophil activation by decreasing the surface expression of both CD35 and CD66b, two markers of neutrophil degranulation. MATERIALS AND METHODS: Venous blood was obtained from three healthy volunteers. Whole blood was incubated with HBSS (control), f-methionyl-leucyl-phenylalanine (fMLP, 1 microM/L), PTX (2 mM/L), or fMLP + PTX. CD35 and CD66b expression were measured by flow cytometry. RESULTS: fMLP treatment caused a significant increase in CD35 and CD66b expression of when compared to controls (P < 0.01). PTX treatment revealed expression of both markers comparable to the control group. A 38% decrease in CD35 (64 +/- 12 versus 100; P < 0.01) and a 52% decrease in CD66b (48 +/- 7 versus 100; P < 0.01) expression were demonstrated in the fMLP + PTX group when compared to fMLP alone. CONCLUSION: In addition to the known effects of PTX on neutrophil oxidative burst, PTX also affects neutrophil degranulation, an essential step in enzyme release and subsequent tissue injury. These findings may have clinical relevance in the treatment of disease processes due to inflammation in which primed neutrophils play a role.

Modulation of lipopolysaccharide-induced acute lung inflammation: Role of insulin.

The present study was undertaken to investigate the influence of insulin on lipopolysaccharide (LPS)-induced acute lung injury. Diabetic male Wistar rats (alloxan, 42 mg/kg, i.v., 30 days) and controls were instilled with saline containing LPS (750 microg/0.4 mL) or saline alone. The following analyses were performed 6 h there after: (a) total and differential cell counts in bronchoalveolar lavage (BAL) fluid, (b) quantification of tumor necrosis factor alpha, interleukin (IL) 1beta, IL-10, and cytokine-induced neutrophil chemoattractant 1 in the BAL (enzyme-linked immunosorbent assay), (c)immunohistochemistry for intercellular adhesion molecule 1 and E-selectin on lung vessels, and (d) quantification of metalloproteinases (MMP) 2 and 9 in the BAL (zymography). Relative to controls, diabetic rats exhibited a reduction in the number of neutrophils (80%) and reduced concentrations of tumor necrosis factor alpha (56%), IL-1beta (66%), and IL-10 (35%) after LPS instillation. Cytokine-induced neutrophil chemoattractant 1 levels did not differ between groups. Increased levels of MMP-2 (90%) and MMP-9 (500%) were observed in diabetic rats compared with controls. Treatment of diabetic rats with neutral protamine Hagedorn insulin (4 IU, s.c.), 2 h before LPS instillation, completely restored the number of neutrophils and concentrations of cytokines in the BAL fluid. Despite no significant differences between diabetic and control groups, there was a remarkable increase in intercellular adhesion molecule 1 and E-selectin expression on lung vessels after insulin treatment. Levels of MMP-2 and MMP-9 did not change after treatment with insulin. Levels of corticosterone were equivalent among groups. Data presented suggest that insulin modulates the production/release of cytokines and the expression of adhesion molecules controlling, therefore, neutrophil migration during the course of LPS-induced acute lung inflammation.

HSPTX protects against hemorrhagic shock resuscitation-induced tissue injury: an attractive alternative to Ringer's lactate.

BACKGROUND: Conventional fluid resuscitation with Ringer's lactated (RL) activates neutrophils and causes end-organ damage. We have previously shown that HSPTX, a combination of small volume hypertonic saline (HS) and pentoxifylline (PTX), a phosphodiesterase-inhibitor, downregulates in vitro neutrophil activation and proinflammatory mediator synthesis. Herein, we hypothesized that HSPTX decreases end-organ injury when compared with RL in an animal model of hemorrhagic shock. METHODS: Sprague-Dawley rats were bled to a mean arterial pressure of 35 mm Hg for 1 hour. Animals were divided into 3 groups: sham (no shock, no resuscitation, n = 7), RL (32 mL/kg, n = 7), and HSPTX (7.5% NaCl 4 mL/kg + PTX 25 mg/kg; n = 7). Shed blood was infused after fluid resuscitation. Blood pressure was monitored until the end of resuscitation. Animals were sacrificed at 24 hour after resuscitation. Bronchoalveolar lavage fluid (BALF) was obtained for white cell count (total and differential) and TNF-alpha and IL-1beta levels were measured by ELISA. Lung and intestinal injury at 24 hour were evaluated by histopathology. Organ damage was graded by a pathologist and a score was created (0 = no injury; 3 = severe). Lung neutrophil infiltration was evaluated by MPO immune staining. RESULTS: There were no differences in mean arterial pressure between groups. At 24 hours, BALF leukocyte count was decreased by 30% in HSPTX animals (p < 0.01). TNF-alpha and IL-1beta levels were markedly decreased in HSPTX-resuscitated animals compared with their RL counterparts (p < 0.01). HSPTX-resuscitated animals (lung injury score = 1.0 +/- 0.4) had markedly decreased acute lung injury compared with RL-treated animals (2.5 +/- 0.3) (p < 0.01). RL resuscitation led to a two-fold increase in lung neutrophil infiltration whereas in HSPTX-treated animals, the number of MPO + cells was similar to sham animals (p < 0.001). Intestinal injury was markedly attenuated by HSPTX (1.1 +/- 0.3) compared with RL animals (2.6 +/- 0.4) (p < 0.001). CONCLUSIONS: HSPTX, a small volume resuscitation strategy with marked immunomodulatory potential led to a marked decrease in end-organ damage. HSPTX is an attractive alternative to RL in hemorrhagic shock resuscitation.

LPS-induced acute lung injury is attenuated by phosphodiesterase inhibition: effects on proinflammatory mediators, metalloproteinases, NF-kappaB, and ICAM-1 expression.

BACKGROUND: Acute endotoxemia is characterized by an enhanced inflammatory response. Pentoxifylline (PTX), a phosphodiesterase inhibitor, has been shown to decrease TNF-alpha levels and to down-regulate neutrophil activation, likely because of increases in intracellular cyclic AMP. Its effects on lipopolysaccharide (LPS) induced lung injury, more specifically on tissue neutrophil infiltration and degranulation, adhesion molecule expression, and transcriptional factor activation, have not been fully investigated. We postulated that PTX treatment in acute endotoxemia downregulates the inflammatory response and may decrease lung injury. METHODS: Male Sprague-Dawley rats were randomized into three groups: Sham (saline i.v.), LPS (5 mg/kg i.v.), and PTX + LPS (25 mg/kg and 5 mg/kg i.v., respectively; concomitant injection). After 4 hours, bronchoalveolar lavage fluid (BAL), plasma, and lungs were sampled. BAL IL-8 (ELISA), BAL MMP-2, plasma MMP-9, and BAL MMP-9 (Zymography) were measured. Lung histology (H&E), in addition to lung MPO, ICAM-1, and NF-kappaB expression evaluated by immunohistochemistry were analyzed. Lung NF-kappaB DNA binding was evaluated by electrophoretic mobility shift assay. RESULTS: PTX treatment decreased BAL IL-8 levels, BAL MMP-2, and plasma MMP-9 activity. Lung neutrophil infiltration (MPO), ICAM-1 expression and NF-kappaB activation were decreased by PTX. In addition, PTX treatment caused a marked attenuation of LPS-induced lung injury. CONCLUSIONS: Phosphodiesterase inhibition by PTX attenuates LPS-induced end-organ injury. In addition, proinflammatory cytokine production is also downregulated, likely because of the marked attenuation of NF-kappaB DNA binding and activation.

Phosphodiesterase inhibition attenuates stored blood-induced neutrophil activation: a novel adjunct to blood transfusion.

BACKGROUND: Activated neutrophils play a central role in the pathogenesis of ARDS and multiple organ failure (MOF). Transfusion of packed red blood cells (PRBCs) is an independent risk factor in the development of ARDS and MOF. It has been postulated that factors present in the supernatant of PRBCs activate neutrophils. The magnitude of neutrophil activation is dependent on the age of the stored blood. Our laboratory and others have reported that pentoxifylline (PTX), a nonspecific phosphodiesterase inhibitor, decreases neutrophil activation. We hypothesized that adding PTX to PRBCs would attenuate blood transfusion-induced neutrophil activation. STUDY DESIGN: Peripheral blood was obtained from healthy human volunteers. Oxidative burst, CD11b, and CD35 expression were measured by flow cytometry using a whole blood preparation. Whole blood was incubated with N-formyl-methionyl-leucyl-phenylalanine (fMLP) (1 microM) alone and 42-day-old PRBC supernatant + fMLP with or without PTX (2 mmol/L). RESULTS: N-formyl-methionyl-leucyl-phenylalanine alone caused a significant increase in neutrophil oxidative burst (100%). The exposure of whole blood to PRBC supernatants + fMLP led to a 1.3-fold increase in neutrophil oxidative burst as compared with fMLP alone, indicating that PRBC supernatants prime neutrophils for oxidative burst by 75%. More importantly, PTX decreased neutrophil oxidative burst by 114% in supernatant + fMLP-stimulated whole blood (p < 0.001). PTX decreased CD11b expression in both fMLP (p < 0.01) and fMLP+supernatant-stimulated whole blood (p < 0.05). Supernatant from PRBCs did not have an additive effect to fMLP alone on CD11b expression. N-formyl-methionyl-leucyl-phenylalanine-induced CD35 expression was downregulated by PTX. The addition of PRBC supernatant did not increase the already upregulated fMLP-induced CD35 expression. CONCLUSIONS: Our results suggest that adding PTX to PRBC supernatant markedly decreases neutrophil activation. The lack of successful treatment strategies to effectively modulate the inflammatory response after blood transfusion indicates the need for novel therapies. Because the deleterious effects of blood transfusion on end-organ injury and MOF are associated with neutrophil activation, the adjunct use of PTX to blood transfusion may have therapeutic potential.

Phosphodiesterase inhibition decreases nuclear factor-kappaB activation and shifts the cytokine response toward anti-inflammatory activity in acute endotoxemia.

BACKGROUND: In sepsis, activation of inflammatory cells and excessive production of proinflammatory cytokines leads to tissue injury, multiple organ failure, and death. We postulated that attenuation but not complete abrogation of hyperinflammation is of clinical benefit in sepsis. Because pentoxifylline (PTX) is known to decrease tumor necrosis factor (TNF)-alpha production and to increase anti-inflammatory cytokine synthesis, we tested the hypothesis that PTX treatment would change the pro- and anti-inflammatory balance and decrease mortality in a murine model of acute endotoxemia. In addition, we investigated the effects of PTX on nuclear factor (NK)-kappaB activation using lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs) as a model. METHODS: Sprague-Dawley rats were treated with intravenous saline (sham), LPS (lipopolysaccharide, Escherichia coli serotype 0111:B4, 5 mg/kg), and concomitant injection of LPS + PTX (25 mg/kg). Four- and 24-hour plasma TNF-alpha and interleukin (IL)-10 levels, 4-hour white cell count, and 24-hour mortality rates were assessed. The IL-10/TNF-alpha ratio was also calculated. Human PBMCs were stimulated with LPS (10 microg/mL) and exposed to PTX (20 mM) concomitantly or 15 minutes after LPS stimulation. I-kappaB phosphorylation by Western blot and NF-kappaB nuclear translocation by electrophoretic mobility shift assay were assessed. RESULTS: PTX markedly down-regulates TNF-alpha production. IL-10 levels at 4 hours were up-regulated in both LPS and PTX + LPS-treated animals; however, levels were higher in the LPS groups, which paralleled high TNF-alpha levels. In contrast, IL-10 levels at 4 and 24 hours in PTX + LPS-treated animals remained constant, whereas in LPS-treated animals, IL-10 levels at 24 hours were markedly decreased. A shift in the internal milieu balance toward anti-inflammatory activity was confirmed by the calculation of the IL-10/TNF-alpha ratio. These changes were not related to changes in the number of circulating leukocytes. The 24-hour mortality rate was 50% in the LPS group and nil in PTX-treated animals. In LPS-stimulated PBMCs, PTX markedly decreases I-kappaB phosphorylation and NF-kappaB nuclear translocation. CONCLUSION: PTX enhances anti-inflammatory activity and decreases mortality in acute endotoxemia. PTX may be an important adjunct to therapies aiming to modulate the inflammatory response in sepsis.

Role of hypertonic saline and pentoxifylline on neutrophil activation and tumor necrosis factor-alpha synthesis: a novel resuscitation strategy.

BACKGROUND: Hypertonic saline (HS) and pentoxifylline (PTX) have been shown to modulate polymorphonuclear neutrophil (PMN) functions after shock and sepsis. We hypothesized that a combination of HS and PTX (HSPTX) would down-regulate PMN functions and inflammatory mediator synthesis more effectively than each alone, possibly by acting at different steps of the signaling pathways, ultimately leading to an enhanced effect. METHODS: Whole blood from healthy volunteers was stimulated with lipopolysaccharide (LPS) (100 microg/mL), f-methionyl-leucyl-phenylalanine (1 micromol/L), and phorbol 12-myristate 13-acetate (1 microg/mL). Baseline oxidative burst was measured by flow cytometry. Two different concentrations of NaCl to achieve increases of 10 mmol/L (HS10) and 40 mmol/L (HS40) above isotonicity, simulating increases in sodium levels seen after infusion of 3% HS and 7.5% HS, were used. PTX (2 mmol/L), HS10, HS40, HSPTX10, and HSPTX40 were added to whole blood concomitantly to the activators. PMN CD14 and CD11b expression were measured by flow cytometry and tumor necrosis factor-alpha levels by enzyme-linked immunosorbent assay in LPS-stimulated whole blood. RESULTS: The combination of PTX with HS10 and with HS40 markedly decreased LPS- (27 +/- 7 and 23 +/- 6 vs. 100; p < 0.01), f-methionyl-leucyl-phenylalanine- (54 +/- 11 and 55 +/- 8 vs. 100; p < 0.05), and phorbol 12-myristate 13-acetate- (30 +/- 4 and 54 +/- 9 vs. 100; p < 0.01 and p < 0.05, respectively) induced PMN oxidative burst. Furthermore, a significant decrease in LPS-induced neutrophil CD11b expression after PTX treatment (79 +/- 5 vs. 100; p < 0.05) and HSPTX40 (68 +/- 7 vs. 100; p < 0.05) was observed. HSPTX10 (7 +/- 0.6; p < 0.001) or HSPTX40 (6 +/- 1.4; p < 0.001) markedly decreased tumor necrosis factor-alpha production to levels similar to those observed with PTX alone (6.5 +/- 0.5; p < 0.001) and significantly lower than HS10 (110 +/- 4.9; p < 0.001) and HS40 alone (83 +/- 1.5; p < 0.001) CONCLUSION: HSPTX down-regulates neutrophil activation and proinflammatory mediator synthesis more effectively that HS alone. HSPTX may have significant applications as a novel fluid resuscitation strategy in hemorrhagic shock.