Outcome of selective non-operative management of penetrating abdominal injuries from the North American National Trauma Database.

BACKGROUND: The aim of this study was to investigate trends in the practice of selective non-operative management (SNOM) for penetrating abdominal injury (PAI) and to determine factors associated with its failure. METHODS: The National Trauma Data Bank for 2002-2008 was reviewed. Patients with PAI were categorized as those who underwent successful SNOM (operative management not required) and those who failed SNOM (surgery required more than 4 h after admission). Yearly rates of SNOM versus non-therapeutic laparotomy (NTL) were plotted. Multivariable regression analysis was performed to identify factors associated with failed SNOM and mortality. RESULTS: A total of 12 707 patients with abdominal gunshot and 13 030 with stab wounds were identified. Rates of SNOM were 22.2 per cent for gunshot and 33.9 per cent for stab wounds, and increased with time (P < 0.001). There was a strong correlation between the rise in SNOM and the decline in NTL (r = - 0.70). SNOM failed in 20.8 and 15.2 per cent of patients with gunshot and stab wounds respectively. Factors predicting failure included the need for blood transfusion (odds ratio (OR) 1.96, 95 per cent confidence interval 1.11 to 3.46) and a higher injury score. Failed SNOM was independently associated with mortality in both the gunshot (OR 4.48, 2.07 to 9.70) and stab (OR 9.83, 3.44 to 28.00) wound groups. CONCLUSION: The practice of SNOM is increasing, with an associated decrease in the rate of NTL for PAI. In most instances SNOM is successful; however, its failure is associated with increased mortality. Careful patient selection and adherence to protocols designed to decrease the failure rate of SNOM are recommended.

Validating the Injury Severity Score (ISS) in different populations: ISS predicts mortality better among Hispanics and females.

INTRODUCTION: The Injury Severity Score (ISS) is the most commonly used measure of injury severity. The score has been shown to have excellent predictive capability for trauma mortality and has been validated in multiple data sets. However, the score has never been tested to see if its discriminatory ability is affected by differences in race and gender. OBJECTIVE: This study is aimed at validating the ISS in men and women and in three different race/ethnic groups using a nationwide database. METHODS: Retrospective analysis of patients age 18-64 y in the National Trauma Data Bank 7.0 with blunt trauma was performed. ISS was categorized as mild (<9,) moderate (9-15), severe (16-25), and profound (>25). Logistic regression was done to measure the relative odds of mortality associated with a change in ISS categories. The discriminatory ability was compared using the receiver operating characteristics curves (ROC). A P value testing the equality of the ROC curves was calculated. Age stratified analyses were also conducted. RESULTS: A total of 872,102 patients had complete data for the analysis on ethnicity, while 763,549 patients were included in the gender analysis. The overall mortality rate was 3.7%. ROC in Whites was 0.8617, in Blacks 0.8586, and in Hispanics 0.8869. Hispanics have a statistically significant higher ROC (P value < 0.001). Similar results were observed within each age category. ROC curves were also significantly higher in females than in males. CONCLUSION: The ISS possesses excellent discriminatory ability in all populations as indicated by the high ROCs.

Differential cytokine expression in skin graft healing in inducible nitric oxide synthase knockout mice.

Inducible nitric oxide synthase (iNOS) and its product, nitric oxide, have been shown to play important roles in wound biology. The present study was performed to investigate the role of iNOS in modulating the cytokine cascade during the complex process of skin graft wound healing.Fifteen iNOS-knockout mice and 15 wild-type C57BL/6J mice were subjected to autogenous 1-cm2 intrascapular full-thickness skin grafts. Three animals in each group were killed on postoperative days 3, 5, 7, 10, and 14. Specimens were then analyzed using nonisotopic in situ hybridization versus mRNA of tumor growth factor-beta1, vascular endothelial growth factor, iNOS, endothelial nitric oxide synthase (eNOS), tumor necrosis factor-alpha, and basic fibroblast growth factor, as well as positive and negative control probes. Positive cells in both grafts and wound beds were counted using a Leica microgrid. Scar thickness was measured with a Leica micrometer. Data were analyzed using the unpaired Student's t test. Expression of iNOS was 2- to 4-fold higher in knockout mice than in wild-type mice on postoperative days 5, 7, and 14. Expression of eNOS was 2- to 2.5-fold higher in knockout mice than in wild-type mice on postoperative days 5 and 7. Tumor necrosis factor-alpha expression was 2- to 7-fold higher in knockout mice than in wild-type mice on all postoperative days. In contrast, expression levels of angiogenic/fibrogenic cytokines (vascular endothelial growth factor, basis fibroblast growth factor, and tumor growth factor-beta1) were 2.5- to 4-fold higher in wild-type mice than in knockout mice. Scars were 1.5- to 2.5-fold thicker in knockout mice than in wild-type mice at all time points. All of the above results represent statistically significant differences (p < 0.05). Significantly different patterns of cytokine expression were seen in knockout and wild-type mice. Although the scar layer was thicker in knockout mice, it showed much greater infiltration with inflammatory cells. These data further delineate the modulatory effect of iNOS and nitric oxide in healing skin grafts.

The role of iNOS in wound healing.

BACKGROUND: We have previously shown that the blockade of nitric oxide (NO) synthesis impairs wound healing, in particular collagen synthesis. Conversely, impaired wound healing is accompanied by decreased wound NO synthesis. Fibroblast collagen synthesis, proliferation, and fibroblast-mediated matrix contraction are critical to wound healing. We examined the wound healing-related phenotypic changes that are induced by the loss of inducible nitric oxide synthase (iNOS) gene function in fibroblasts. METHODS: Dermal fibroblasts were obtained from 8- to 12-week-old iNOS--knock out (KO; C57BL/Ai-[KO] Nos2 N5) and wild type mice by an explant technique and used after 1 to 3 passages. Proliferation ([(3)H]-thymidine incorporation) and collagen synthesis ([(3)H]-proline incorporation into collagenase-sensitive protein) were studied after stimulation with 10% fetal bovine serum. Matrix remodeling was assessed by the measurement of the contraction of fibroblast-populated collagen lattices. RESULTS: iNOS-KO fibroblasts proliferated more slowly, synthesized less collagen, and contracted fibroblast-populated collagen lattices more slowly than wild-type fibroblast. Collagen synthesis was restored to normal in KO fibroblasts in response to NO donors (s-nitroso-N-acetylpenicillamine). CONCLUSIONS: iNOS deficiency causes significant impairment in wound healing-related properties of fibroblasts, which suggests that NO plays an important role in wound healing.

Laparoscopic surgery in older adults.

This article provides an overview of the current role of laparoscopic surgery in older patients. A retrospective review and analysis of the recent English-language literature on laparoscopic procedures with special attention devoted to those articles focused on geriatric patients was performed. Laparoscopic surgery has rapidly become the fastest-growing discipline within the surgical arena and new applications for laparoscopy continue to be reported. The primary benefits to patients of these developments are smaller scars, decreased postoperative pain, and more-rapid return to normal activity. As society ages, more older patients will present with pathology amenable to laparoscopic intervention. Several aspects of laparoscopy impose unique physiologic stresses and, as such, may alter surgical risk to the geriatric patient. In addition, older patients often have delayed surgical interventions because of more-conservative medical management or unusual symptomatology, which may further complicate the laparoscopic approach. These limitations may alter the risk-to-benefit ratio of laparoscopic versus open procedures. Despite this lack of elucidation of full-risk profiles, laparoscopic approaches should be considered regardless of a patient's age.

A novel method of studying wound healing.

In order to study wound healing, it is often necessary to administer various wound-active substances by the systemic route. It is unclear whether the observed effects are the result of local or systemic influence of the agent administered. Furthermore, high systemic doses are often required to achieve activity at the wound level. Direct intrawound administration of substances is traumatic and disruptive to the fragile wound environment and increases the risk of infection. We devised a system for continuous atraumatic delivery of substances directly to subcutaneously implanted polyvinyl alcohol sponges, an adaptation of a well-established model of wound healing. Sponge-catheter constructs were fashioned by feeding identical lengths of silicone catheters through two 40-mg sponge disks (on edge). The distal sponge was fixed 0.5 cm from the distal, ligated end of the catheter and centered over two 1-mm holes in the catheter tubing. The proximal sponge was fixed over nonperforated catheter with its edge 2 cm proximal from the close edge of the distal sponge. Each construct was connected to a mini-osmotic pump (infusion rate 1 microl/h) loaded with an appropriate infusate and inserted subcutaneously on the dorsum of anesthetized male Sprague-Dawley rats. Hydroxyproline (OHP) content of sponges, a measure of collagen deposition, was determined at 7 days postwounding. Infusion of India ink confirmed selective delivery to the distal sponge. Saline infusion alone significantly elevated OHP content compared to noninfused sponges (450 +/- 43 vs 328 +/- 36 microg OHP/100 mg sponge, P < 0.05). Infusion of S-methylisothiourea (a selective iNOS inhibitor, 84 microg/sponge/24 h) successfully inhibited NO production (35.9 +/- 3.1 vs 49.6 +/- 3.6 microM, P < 0.05) and decreased sponge OHP content (385 +/- 60 vs 568 +/- 70 microg OHP/100 mg sponge, P < 0.05) without the toxic side effect (i.e., weight loss) seen with systemic administration. Infusion of an adenoviral solution containing mouse iNOS cDNA resulted in successful transduction of wound cells demonstrating the ability to deliver genes to a healing wound model. The data demonstrate that manipulation of wound physiology is possible by local delivery of low doses of wound-active compounds to the wound site. This promises to be a powerful tool for the study of both normal and impaired wound healing.

Effect of matrix metalloproteinase inhibition on colonic anastomotic healing in rats.

Wound strength depends on the balance between collagen synthesis and degradation; however, the role of collagen breakdown in wound healing is still not well understood. We investigated the role of matrix metalloproteinases in wound healing by using BE16627B, a matrix metalloproteinase inhibitor. Identical surgical procedures consisting of a colonic anastomosis (single-layer, inverted) and implantation of an osmotic pump in the back were performed in male Sprague-Dawley rats weighing 270 to 290 grams. The animals were randomly assigned to receive either BE16627B (n = 10) dissolved in dimethylsulfoxide and diluted with ethylene glycol at a dosage of 2.4 mg/rat/day for 3 days or the vehicle solution alone (n = 11). The solutions were administered through the surgically implanted osmotic pumps. The animals were killed 4 days after surgery, and the colonic bursting pressure (mm Hg) and hydroxyproline concentration (microg/mg wet tissue, index of collagen) were measured. The administration of BE16627B enhanced colonic anastomotic healing, as measured by the increase in the colonic bursting pressure (160 +/- 12 vs. 125 +/- 7 mm Hg; P < 0.05) and the increase in the soluble fraction of collagen (0.27 +/- 0.01 vs. 0.21 +/- 0.01 microg/mg wet tissue; P < 0.01) in the anastomosis. Histologic examination of the tissue revealed that the use of BE16627B resulted in the preservation of the multilayered colonic structure and increased the network of collagen between both ends of the colon in the thickening submucosal layer. These findings demonstrate that the inhibition of matrix metalloproteinase activity influences colonic anastomotic healing, indicating a potential mechanism for enhancing anastomotic healing.

Trauma and wound healing: role of the route of nutritional support.

Following severe trauma and surgical injury, nutritional support via the enteral route has been shown to lead to increased survival and decreased complications when compared to the parenteral route. We hypothesized that the route of nutrient delivery may affect cutaneous wound healing following severe traumatic insult. Forty-six Sprague Dawley rats underwent bilateral closed femoral fractures, central venous catheterization, gastrostomy placement and dorsal skin incision with placement of polyvinyl alcohol sponges into subcutaneous pockets. Identical nutritional infusates of 25% dextrose, 4.25% amino acids, and vitamins were given, half the animals receiving the infusion via the gastrostomy (ENT) and the other half via the venous catheter (TPN). Animals were sacrificed on post-operative days 5, 7, or 10. Wound breaking strength (WBS, g) and sponge granuloma hydroxyproline content (OHP- a measure of wound collagen deposition, microg/ 100mg sponge) were measured. There were no significant nutritional differences between the two feeding groups. On days 5 and 7, WBS was significantly higher in the ENT group (58.0 +/- 3.1 g vs 48.9 +/- 2.6 g, p < 0.05, and 123 +/- 19 g vs 87.6 +/- 4.2 g, p < 0.05 vs TPN respectively). Sponge OHP content on day 5 was significantly higher in the ENT group (101 +/- 3 vs 86.7+/-5.8 microg/100 sponge, p < 0.05). These data demonstrate that the enteral feeding route imparts a benefit to early post-traumatic wound healing s compared to parenteral feeding.

Supplemental dietary arginine enhances wound healing in normal but not inducible nitric oxide synthase knockout mice.

BACKGROUND: Although generation of nitric oxide (NO) from inducible nitric oxide synthase (iNOS) has been shown to be required for cutaneous wound healing, no differences have been noted in incisional healing between iNOS knockout (iNOS-KO) and wild type (WT) mice. Because supplemental dietary arginine enhances cutaneous healing in normal rodents and is the sole substrate for NO synthesis, we studied whether arginine can enhance cutaneous wound healing in iNOS-KO mice. METHODS: Twenty iNOS-KO and 20 WT mice, all on a C57BL/6 background, were divided into 4 groups of 10 animals each. Ten animals with each trait were randomized to receive either normal food and tap water or food and water each supplemented with 0.5% arginine (w/w). All animals underwent a 2.5-cm dorsal skin incision with implantation of four 20-mg polyvinyl alcohol sponges into subcutaneous pockets. On postoperative day 14 the animals were killed. The dorsal wound was harvested for breaking strength determination and the wound sponges were assayed for hydroxyproline content and total wound fluid nitrite/nitrate concentration. RESULTS: Dietary arginine supplementation enhanced both wound breaking strength and collagen deposition in WT but not iNOS-KO mice. Wound fluid nitrite/nitrate levels were higher in WT than iNOS-KO animals but were not significantly influenced by additional arginine. CONCLUSIONS: These data demonstrate that supplemental dietary arginine enhances wound healing in normal mice. The loss of a functional iNOS gene abrogates the beneficial effect of arginine in wound healing. This suggests that the metabolism of arginine via the NO pathway is one mechanism by which arginine enhances wound healing.

Role of nitric oxide in wound healing.

Nitric oxide is a short-lived free radical, that is capable of multiple effects at the molecular, cellular, and physiologic levels. Over the past several years, nitric oxide has been proved to play an important role in the healing of various types of wounds. The present review examines some of the recently defined roles of nitric oxide in normal and pathologic healing.

Inhibition of tumor necrosis factor-alpha attenuates wound breaking strength in rats.

Exogenous administration of tumor necrosis factor-alpha has been shown to both enhance and attenuate cutaneous healing in a dose-dependent manner. We examined the effects of tumor necrosis factor inhibition in the healing wound by both systemic and local administration of tumor necrosis factor-binding protein. Male Balb/C mice underwent dorsal skin incision with subcutaneous implantation of 20 mg polyvinyl alcohol sponges (4 per animal). In Experiment I, one group (n = 20) received intraperitoneal injections of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding, while another group (n = 20) received saline. Four animals from each group were euthanized on days 1, 3, 5, 7, and 14 postwounding. In Experiment II, one group (n = 10) received an intraperitoneal injection of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding and every third day thereafter. Another group (n = 10) received an intraperitoneal injection of saline at the time of wounding and every third day thereafter. In Experiment III, one group received a single intraperitoneal injection of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding (n = 7), or on postwounding day 4 (n = 7), or day 7 (n = 7). Another group received saline injections at the time of wounding (n = 7), or on postwounding days 4 or 7 (n = 7, respectively). All animals in Experiments II and III were killed at postwounding day 14. Wound breaking strengths were assessed using a tensiometer. Wound fluid collected from the implanted sponges was assayed for tumor necrosis factor-alpha and tumor necrosis factor-binding protein levels using a biological assay and enzyme-linked immunosorbent assay, respectively. Collagen gene expression in sponge granulomata was assessed by Northern analysis. Collagen deposition in sponges was quantified by measuring hydroxyproline content. Wounds were significantly weaker in the animals that received repeated injections of tumor necrosis factor-binding protein with a mean wound breaking strength of 93.1 g vs. 186.6 g in controls (p < 0.05). Wound breaking strength in groups that received a single injection of tumor necrosis factor-binding protein on either day 0, 4, or 7 postwounding were no different than their respective controls. There was no difference in the mean hydroxyproline content of sponges between any of the tumor necrosis factor-binding protein groups and their respective controls. Northern analysis for collagen I and III expression also revealed no differences. These data indicate that continued systemic administration of tumor necrosis factor-binding protein resulted in significantly weaker wounds with no corresponding differences in wound collagen content, and collagen gene expression. This suggests that tumor necrosis factor-alpha inhibition throughout healing leads to a qualitatively impaired wound without a quantitative alteration in collagen deposition.

Enhancement of fibroblast collagen synthesis by nitric oxide.

Fibroblasts can be stimulated by cytokines to synthesize nitric oxide (NO, nitrogen monoxide), while wound-derived fibroblasts synthesize NO spontaneously. Since wound fibroblasts are phenotypically characterized by greater collagen synthesis when compared to fibroblasts derived from noninjured tissue, we hypothesized that there may be a correlation between wound-induced NO synthesis and enhanced collagen production. To study the role of NO on collagen metabolism, normal dermal fibroblasts were cultured in the presence or absence of the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) and their collagen metabolism was studied on the transcriptional as well as translational level. Fibroblast collagen synthesis was enhanced by 74.3 +/- 18.2 and 87.5 +/- 28.2% in the presence of 100 and 400 microM SNAP, respectively. This effect was not due to increased collagen type I or type III gene transcription. Cellular proliferation measured by thymidine incorporation was significantly decreased in the presence of SNAP, indicating that the increased collagen production was due to a net increase of collagen synthesis by the cells. Investigation of the collagen breakdown pathway showed that neither collagenase gene expression nor collagenase protein expression was affected by SNAP. The results of this study demonstrate for the first time that NO enhances collagen synthesis, most likely at a posttranslational level.

Expression and function of inducible nitric oxide synthase during rat colon anastomotic healing.

Nitric oxide plays a significant but incompletely understood role in fibroblast function and cutaneous wound collagen synthesis; however, the participation of inducible nitric oxide synthase (iNOS) in gastrointestinal anastomotic healing has not been studied. Male Sprague-Dawley rats underwent single-layer left colonic anastomosis. Animals were killed at 24-hour intervals postoperatively and the anastomosis was excised. Parallel uninjured colon tissue samples were also analyzed. Reverse transcriptase-polymerase chain reaction confirmed the absence of iNOS messenger RNA in control colon and expression of the gene in anastomotic tissue on all study days. Northern hybridization demonstrated maximal iNOS messenger RNA transcription on day 1 with decreased levels on days 3 and 5. iNOS enzyme activity, measured biochemically by the conversion of [(3) H-arginine to [(3) H]-citrulline ex vivo, was also maximal on day 1 (7.35 +/- 1.34 pmol/mg protein/min [+/- standard error of the mean], n = 10) and decreased on days 3 (4.37 +/- 2.32 pmol/mg protein/min; n = 6) and 5 (2.80 +/- 0.92 pmol/mg protein/min; n = 6). Immunohistochemical staining demonstrated that (1) iNOS expression is confined to a discrete cell population in the region of the anastomosis containing inflammatory cells; (2) those cells assume a highly conserved position on the luminal edge of the proliferating scar; and (3) the iNOS-expressing cells are present throughout the fibroplastic phase of healing. To functionally assess the role of iNOS in colonic healing, rats were treated with a continuous intravenous infusion of S-methylisothiourea (a selective inhibitor of iNOS) at a dosage of 200 mg/kg/day for 5 days after anastomosis. There was a significantly reduced anastomotic bursting pressure in rats treated with the inhibitor as compared to rats treated with intravenous normal saline solution (108.4 +/- 13.2 mm Hg vs. 148.4 +/- 10.3 mm Hg; P <0.05). These results suggest that iNOS gene expression is induced during colonic anastomotic healing, that it is present through all phases of healing but is maximal through the inflammatory phase, and that iNOS activity is required for optimal anastomotic healing.

Effect of nutritional route on colonic anastomotic healing in the rat.

Although early enteral feeding has been shown to benefit cutaneous healing when compared to parenteral feeding, the effect of the route of nutritional support in gastrointestinal anastomotic healing has not been defined. The aim of the present study was to determine whether the route of nutritional support influences colonic anastomotic healing. Twenty male Sprague-Dawley rats weighing 270 to 290 grams underwent identical surgical manipulation consisting of central venous catheterization, gastrostomy insertion, and distal colonic anastomosis (single-layer, inverted). Identical nutrient infusates composed of 4.25% amino acids, 25% dextrose, and vitamins were administered, with half the animals receiving the infusions via the gastrostomy and the other half via the venous catheter. Animals were killed 5 days after surgery. There were no differences in nutritional parameters between the parenterally and enterally fed groups. Colonic anastomotic bursting pressure was significantly higher in the enterally fed group (180 +/- 6 vs. 150 +/- 11 mm Hg; P <0.01). The measured insoluble collagen and total protein content in anastomotic tissue were enhanced in the enterally supported group. The fraction of soluble (newly synthesized) collagen did not differ between the two groups. The data demonstrate that the route of nutrient administration influences colonic anastomotic healing. The preservation of colonic structural collagen in the enteral group may improve the ability of the gut to hold sutures and thus enhance anastomotic healing.

Arginine and nutrition in renal disease.

The physiological significance of arginine metabolism extends far beyond its incorporation as an amino acid into proteins. In addition to its effects when administered as a dietary supplement, the end-products of arginine metabolism by the enzymes arginase, arginine decarboxylase (ADC), and nitric oxide synthase (NOS) have been shown to play roles in wound healing, immune response, tumor biology, and the regulation of inflammation. These properties make arginine metabolism a significant concern in defining and, likely, treating renal disease.

Metalloproteinase inhibitors and wound healing: a novel enhancer of wound strength.

BACKGROUND: Wound strength is a balance between collagen synthesis and degradation. The role of collagen breakdown in wound healing is still not well understood. We investigated the role of collagenases (metalloproteinases [MMPs]) in wound healing in using GM6001, a novel inhibitor of MMPs. METHODS: We used the dorsal skin incision model with implantation of polyvinyl alcohol sponges. Twenty male Sprague-Dawley rats were randomly assigned to receive either GM6001 (10 mg/kg body weight) or 2 mL saline subcutaneously. Ten days after operation the animals were killed and fresh wound breaking strength, scar and sponge hydroxyproline content, and collagen type I gene expression in sponges were assayed. In addition, the inflammatory response and the wound fluid cytokine (tumor necrosis factor-alpha [TNF-alpha] and transforming growth factor-beta 1 [TGF-beta 1]) profile were studied. RESULTS: GM6001 significantly increased wound strength (422 +/- 59 vs 302 +/- 33 g, P < .05), whereas scar collagen content did not differ. In the sponge granulomas the inflammatory infiltrate, the collagen content, and the collagen type I gene expression were all significantly decreased by GM6001. CONCLUSIONS: Inhibition of MMP activity during acute wound healing enhances wound strength even though new collagen synthesis and the inflammatory response are significantly decreased. This could be achieved by decreasing collagen turnover or increasing collagen maturation and crosslinking, or both.

Modulation of inflammation and immunity by arginine supplements.

Arginine holds a key position in the cellular functions and interactions that occur during inflammation and immune responses. The competition for arginine as a substrate between nitric oxide synthase and arginase appears to be at the core of the regulation of the inflammatory process. This review examines some of the recently defined effects of arginine on various inflammatory processes and immune cell functions.

Nitric oxide generation from L-arginine is required for optimal human peripheral blood lymphocyte DNA synthesis.

We examined whether L-arginine is a substrate for nitric oxide (NO) production by peripheral blood mononuclear cells (MNC) in vitro. Minimal extracellular arginine (0.04 mmol/L) is required for maximal lymphocyte proliferation after phytohemagglutinin stimulation. In the absence of arginine, proliferation was 41% of normal without loss of viability. In contrast, MNC total protein synthesis (as assessed by tritiated leucine incorporation) or lymphokine synthesis (interleukin-2, as assessed by cytotoxic lymphoid line (CTLL) proliferation) were not affected by the absence or presence of arginine in the medium. Exogenous nitric oxide provided as sodium nitroprusside could replace L-arginine for maximal blastogenic proliferation. The addition of NG-monomethyl-L-arginine (NMMA; 0.1 mmol/L), a specific inhibitor of the NO synthetic pathway, significantly reduced DNA synthesis both at 0 and 0.1 mmol/L arginine concentrations; this effect was reversed to 91% of normal by excess arginine (1.0 mmol/L). Homoarginine (0.1 mmol/L; a known substrate for NO production) partially substituted for arginine, and this effect was also abrogated by NMMA. Nitrite levels (an end product of NO metabolism) were reduced when L-arginine was absent or NMMA was added to L-arginine-containing media. Cytosol from phytohemagglutinin-stimulated MNC-enhanced cyclic guanosine monophosphate production in the presence of L-arginine as substrate. The data suggest that the inductive effects of L-arginine on MNC DNA synthesis are not related to its nutrient requirement for protein synthesis, but rather caused by its role as a substrate for NO production. MNC actively synthesize NO during mitogenic proliferation. NO appears to be a promoter of MNC DNA synthesis, probably by its well-known effect as an activator of guanylate cyclase, which increases cyclic guanosine monophosphate levels.