Robert P. Hummel

Comparison of postoperative wound complications and early cancer recurrence between patients undergoing mastectomy with or without immediate breast reconstruction

The incidence of postoperative wound complications and early cancer recurrence was studied in 289 patients who had mastectomy alone and in 113 patients who underwent immediate reconstruction following mastectomy. Patients undergoing immediate reconstruction were younger and had less advanced disease than patients who had mastectomy alone. The postoperative hospital stay was 3.8 days and 4.4 days (p < 0.05) in patients with and without reconstruction, respectively. The overall incidence of postoperative complications was similar in the two groups of patients: 31% and 28% in patients with and without reconstruction, respectively. The incidence of postoperative seroma was higher among patients with mastectomy alone (19% versus 3%, p < 0.05), whereas the incidence of other wound complications was similar in the two groups of patients. Prosthesis-specific complications occurred in 17%. Eight prostheses were removed because of complications. During the relatively short follow-up period (approximately 20 months), local recurrence was noted in 16 patients (6%) who had mastectomy alone and in 1 patient (1%) who had immediate reconstruction after mastectomy (p < 0.05). There was no significant difference in the incidence of distant metastases between the two groups of patients. The results suggest that immediate breast reconstruction can be performed following mastectomy for cancer without increased risk for overall postoperative complications, prolonged hospital stay, or local recurrence. However, patients who choose to have immediate reconstruction need to be informed about risks for specific complications associated with the procedure, especially if an implant is used.

Synergistic actions of platelet‐derived growth factor and the insulin‐like growth factors in vivo

Topical application of growth factors has been shown to benefit both normal and impaired wound healing. In normal tissue repair, resident cells produce a “cocktail” of various types of growth factors that overlap in function. In vitro studies have proved that growth factor combinations can act synergistically to enhance cellular function beyond that achieved with individual growth factors. To determine whether similar combinations have a synergistic effect in vivo, we applied growth factor combinations topically to full‐thickness skin wounds created in genetically diabetic mice. The C57BL/KsJ‐db/db mouse is obese and has insulin‐resistant diabetes, and it has been proved that this mouse has markedly impaired wound healing. Topical application of platelet‐derived growth factor, insulin‐like growth factor‐I, or insulin‐like growth factor‐II enhances healing in this model. Marked synergism was found when platelet‐derived growth factor and insulin‐like growth factor‐II were combined to produce augmentation in wound closure beyond that achieved by application of the individual growth factors. The synergistic effect allowed for improved tissue repair at doses of platelet‐derived growth factor and insulin‐like growth factor‐II that were ineffective when applied individually. The addition of insulin‐like growth factor‐I or insulin to platelet‐derived growth factor produced no significant synergism. Because multiple growth factors are released in the wound during the healing process, it is not surprising that their combination further enhances healing. Growth factor combinations should become an important addition to the armamentarium for the treatment of chronic, nonhealing wounds.

Inhibition ofCandida albicans by Escherichia coli in vitro and in the germfree mouse

1.Candida albicans readily establishes in the gastrointestinal tract of germfree mice either alone or in the presence ofE. coli. 2. Noninhibitory and inhibitory groups ofE. coli have been separated by their actionin vitro againstC. albicans. 3. The inhibitoryE. coli significantly lowers the levels ofC. albicans in stools of gnotobiotic mice. A dialysate of the inhibitoryE. coli also lowers theCandida levels in the stool of other gnotobiotic mice. 4. Gentamicin therapy and a 10% third degree burn had no effect on theCandida levels in the stool of germfree mice nor did it cause invasive infection or mortality in the animals.

The effect of sepsis in rats on skeletal muscle protein synthesis in vivo and in periphery and central core of incubated muscle preparations in vitro

Recent studies demonstrated the development of a central, hypoxic core in incubated rat skeletal muscles. The influence of a central core on changes in protein synthesis rate, observed in incubated muscles from septic rats, is not known. In the present study, intact soleus muscles from 40 to 60-g sham-operated control rats and from septic rats (16 hours after cecal ligation and puncture) were incubated in vitro in a flaccid or stretched state. Protein synthesis rate was determined in whole muscle and in the central core and periphery of the muscle by measuring incorporation of 14C-phenylalanine into protein. Protein synthesis rate in vivo was measured with a flooding-dose technique using 3H-phenylalanine. The development of a central, hypoxic core in incubated muscles was assessed histochemically by staining the muscles for alpha-glucan phosphorylase activity. A central core with loss of alpha-glucan phosphorylase activity was noted after incubation for 30 minutes in both control and septic muscles. The protein synthesis rate was lower in the central core than in the periphery of incubated flaccid control muscles. In all other in vitro muscle preparations, however, there were no significant differences in protein synthesis rate among whole muscles, central core and periphery. Protein synthesis rate in septic muscles was reduced to a similar extent, approximately 20%, in vivo and in the different in vitro preparations, both when measured in whole muscle and in the central core or periphery.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of catabolic hormone infusion on protein turnover and amino acid uptake in skeletal muscle

Increased plasma levels of the catabolic hormones glucagon, epinephrine, and cortisol have been implicated in mediating various metabolic alterations in trauma and sepsis. Their role in altered protein turnover and amino acid transport in skeletal muscle during sepsis, however, is not known. In the current study, rats were infused with a mixture of the catabolic hormones for 16 hours. Control animals were infused with vehicle solution. Protein synthesis and degradation rates were measured in incubated, intact soleus muscles as incorporation of 14C-phenylalanine into protein and release of tyrosine into incubation medium, respectively. Muscle amino acid uptake was determined by measuring the intracellular to extracellular ratio of [3H]-alpha-aminoisobutyric acid after incubation for 2 hours. Infusion of catabolic hormones for 16 hours resulted in elevated plasma glucose and lactate levels, reduced plasma concentrations of most amino acids, and accelerated muscle protein breakdown, similar to previous findings in septic rats. Protein synthesis rates and amino acid uptake in incubated muscles were not significantly different in control and hormone-infused rats. The current study suggests that increased muscle proteolysis in sepsis and severe injury may be mediated in part by catabolic hormones. In contrast, reduced muscle protein synthesis and amino acid uptake are probably signaled by other substances or mechanisms.

Effect of sepsis on amino acid transport system A and its response to insulin in incubated rat skeletal muscle

The effect of sepsis on neutral amino acid transport systems A, ASC, and L, was studied in incubated rat soleus (SOL) muscles. We also examined the effects of plasma from septic rats and of varying concentrations of insulin (10 to 10(5) microU/mL), added in vitro to incubated muscles, on system A amino acid transport. Sepsis was induced by cecal ligation and puncture (CLP) in rats weighing 40 to 60 g. Control rats were sham-operated. System A activity was assessed by determining uptake of 2-(methylamino)isobutyrate (MeAIB) 16 hours after CLP or sham-operation. System ASC was studied by measuring uptake of alpha-aminoisobutyric acid (AIB) in the presence of 25 mmol/L MeAIB and 25 mmol/L 2-amino-2-norbornane carboxylic acid (BCH) to inhibit uptake by systems A and L. System L activity was defined as sodium-independent uptake of cycloleucine. MeAIB uptake was reduced by 28% in muscles of septic rats, while amino acid transport by systems ASC and L was almost identical in muscles from control and septic rats. Addition of plasma from septic rats to incubated normal SOL muscles inhibited MeAIB uptake by 31%. Addition of insulin to the incubation medium resulted in increased uptake of MeAIB, both in nonseptic and septic muscle. The lowest hormone concentration tested that significantly enhanced MeAIB uptake in nonseptic muscle was 10(2) microU/mL and in septic muscle 10 microU/mL. The results suggest that sepsis in rats specifically inhibits amino acid transport system A and that reduced muscle amino acid uptake may be caused by a circulating factor in sepsis.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of sepsis and starvation on amino acid uptake in skeletal muscle.

Sepsis and starvation are catabolic conditions often occurring simultaneously. We recently found that uptake of the nonmetabolized amino acid analog, alpha-aminoisobutyrate (AIB), was reduced in skeletal muscle during sepsis, and data were presented suggesting that this reduction was caused by a factor present in septic plasma. The purpose of the present study was to determine the effect of starvation on amino acid uptake and to investigate possible interactions between sepsis and starvation regarding effects on muscle amino acid uptake. Soleus muscles were removed intact from rats fasted for 0, 24, 48, or 72 hr. Muscles were incubated for 2 hr in Krebs-Henseleit bicarbonate buffer (pH 7.4) and glucose (5 mM) with or without 0.25 ml of septic plasma (total volume 3 ml). Muscle AIB uptake progressively decreased by approximately 50% during the first 48 hr of starvation, with no further decrease at 72 hr. Addition of septic plasma in vitro reduced AIB uptake in muscles from fed rats and from rats which had been fasted for 24 hr, but did not further reduce uptake after longer periods of starvation. The results suggest that starvation reduces amino acid uptake in skeletal muscle and that the nutritional status of muscle is an important component of the response to the factor(s) in septic plasma capable of inhibiting amino acid uptake.

Increased blood-brain transport of tryptophan after portacaval anastomoses in germ-free rats.

Portal-systemic shunting in rats results in altered plasma concentrations of the large neutral amino acids (NAA), and increased blood-brain barrier NAA transport. Bacterial ammonia production in the bowel, especially the colon, is thought to play a major role in the etiology of portal-systemic encephalopathy (PSE). In order to isolate the role of bacteria in PSE, plasma and brain amino acids and ammonia (NH3) were studied in germ-free (GF) rats with portacaval shunts (PCS). Germ-free rats underwent end-to-side portacaval shunt or sham operation under germ-free conditions and were kept in a germ-free isolator under careful bacteriologic control. Similar operations were carried out on conventional animals. Two weeks post-operatively blood-brain transport of the neutral amino acid tryptophan was studied and the animals sacrificed. Plasma NH3 rose after PCS both in GF and in conventional rats. In germ-free and conventional rats with PCS, there was a significant elevation in plasma of the aromatic amino acids and decreased concentrations of the branched-chain amino acids. In the brain, both groups had elevated aromatic amino acids, while the branched-chain amino acids remained normal. The blood-brain transport of tryptophan was elevated in portacaval shunted rats, whether germ-free or conventional. These studies suggest the elevated plasma levels of ammonia and disturbances in plasma and brain amino acids seen after portacaval shunt are not dependent on the presence of gut bacteria.

Inhibited amino acid uptake in skeletal muscle during starvation.

Amino acid uptake in skeletal muscle is reduced during different catabolic conditions such as sepsis, endotoxic shock, and uremia. The present study was designed to determine the effect of another catabolic condition, starvation, on amino acid transport in skeletal muscle. Male Sprague-Dawley rats (40-60 g) were starved for 24, 48, or 72 hr and soleus (SOL) muscles were removed intact and incubated for 2 hr in a medium consisting of Krebs-Henseleit bicarbonate buffer (pH 7.4) with glucose (5 mM), [14C]-inulin, and [3H]-alpha-aminoisobutyric acid (AIB). Amino acid uptake was determined from intracellular to extracellular ratio of AIB following incubation. AIB uptake was significantly reduced after 24 hr of starvation and remained low with further fasting. After 72 hr the AIB distribution ratio was approximately 50% of initial value. Amino acid uptake returned to normal within 24 hr after refeeding of animals that had been starved for 72 hr. Plasma (0.25 ml) from starved rats, added to the incubation medium (2.75 ml) of muscles from nonfasted rats, significantly inhibited AIB uptake. The present results suggest that amino acid uptake in skeletal muscle is progressively reduced during starvation, an effect that may be mediated by a circulating factor(s) present in blood.

Effects of indomethacin and leupeptin on muscle cathepsin B activity and protein degradation during sepsis

The roles of prostaglandins and lysosomal proteases in accelerated skeletal muscle proteolysis during sepsis are not yet fully understood. In this study rats received intraperitoneal injections of the prostaglandin synthesis inhibitor indomethacin (IND, 5.0 mg/kg), the lysosomal cathepsin B inhibitor leupeptin (LEU, 2.5 mg/kg), or normal saline 2 hr before cecal ligation and puncture (a model of intraabdominal sepsis) or sham-operation. The injections were repeated every 6 hr for a total of four doses. Sixteen hours after operation, intact extensor digitorum longus (EDL) muscles were harvested and cathepsin B activity was measured in one muscle. The contralateral muscle was incubated in oxygenated Krebs-Henseleit bicarbonate buffer containing glucose (10 mM) and cycloheximide (0.5 mM), and protein degradation rate was determined as the release of tyrosine into the incubation medium. Both muscle cathepsin B activity and protein degradation rate were higher in septic than in sham-operated rats. Treatment with IND or LEU significantly reduced the elevated cathepsin B activity in septic muscles, but failed to significantly alter muscle proteolysis. In nonseptic muscle, both cathepsin B activity and protein degradation rate were unaffected by the different types of treatment. The results suggest that although prostaglandins may influence muscle lysosomal protease activity, neither prostaglandins nor the lysosomal protease cathepsin B appear to be major regulators of accelerated muscle protein breakdown during sepsis.