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The mitotic activity of blood monocytes cultured in autologous wound fluid.

Wound fluid was obtained from the subcutaneous tissue of sheep by implanting a perforated Teflon sampling chamber. A catheter from this chamber was brought to the surface. The types and numbers of cells in this fluid were recorded. Cell free fluid was used at either 100% concentration, or 25% concentration in plasma, as a medium for the culture of autologous blood monocytes. Its ability to support mitosis was compared with that of 100% autologous plasma. Wound fluids taken upon consecutive post-implantation days were compared. Cultured monocytes survived in small numbers in wound fluid collected on the days immediately following operation. When mitosis occurred the mitotic indices remained low. The wound fluid did not exhibit that mitogenicity which is apparent in autologous plasma taken on the third or fourth days following implantation. The results suggest that, following the implantation of Teflon, the wound fluid contains a component which is toxic for blood monocytes cultured upon glass.

Animals

Antimicrobial systems of the surgical wound. II. Detection of antimicrobial protein in cell-free wound fluid.

Human wound fluid contains heat-stable proteins with moderate antibacterial activity against Staphylococcus aureus and Escherichia coli, and different heat-labile proteins with antibacterial activity against E coli. Blood serum also contains heat-labile antibacterial substances, but little heat-stable activity against Staphylococcus aureus. Both blood serum and wound fluid have bacteriostatic activity against S epidermidis, and early growth of Streptococcus fecalis occurs in serum and wound fluids. The concentration or activity of antimicrobial proteins increases during the first week in the fresh wound and then decreases as the wound matures.

Blood Bactericidal Activity

Effect of a deproteinized blood extract on experimental granulation tissue.

The present work was undertaken to study the effects of a deproteinized extract of calves' blood (Solcoseryl) on developing granulation tissue in rats. Cylindrical hollow viscose cellulose sponge implants were used as an inductive matrix for the growth of granulation tissue. In the first, sham group the implants were treated daily by withdrawing 1 ml of wound fluid from the central dead space of the implant and then re-injecting the fluid. In the second, experimental group the aspirated wound fluid was replaced by a corresponding volume or Solcoseryl. Analyses of wound fluid and granulation tissue were carried out 4, 10 and 21 days after implantation. A statistically significant increase of granulation tissue hemoglobin (+21%) was observed at 10 days in the Solcoseryl group as compared with the sham-treated rats, indicating an enhanced capillary ingrowth. Concurrently, the mean amount of DNA in the Solcoseryl-treated tissues was elevated by 48% over the level of the sham-treated group, demonstrating an augmented cellularity of granulation tissue. At 21 days the mean amount of collagen hydroxyproline of the Solcoseryl group was 31% above the level measured in the sham-treated animals. PO2, PCO2 and pH in the wound fluid and the amounts of RNA and uronic acids showed no essential differences between the two groups. These data demonstrate a stimulatory effect of Solcoseryl on several aspects of granulation tissue formation: augmented vascularization, elevated cellularity and subsequent enhancement in the accumulation of collagen.

Animals

Quantitation of local acidosis and hypoxia produced by infection.

Increased strength of healing incisions infected with E coli was demonstrated in this experiment. Efforts to measure respiratory gas tensions and pH in these incisions were unsuccessful. Therefore, these moieties were measured in normal and infected wound fluid contained in implanted wire cylinders. The wound fluid from infected cylinders was consistently more acidotic and had a lower pO2 and a higher pCO2 than fluid from unifected wound cylinders.

Animals

Prostaglandins and the sources of their production in a mild inflammatory lesion in sheep.

1. Wound fluid was collected from a subcutaneous lesion in the shoulder region of sheep by implanting a PTFE sampling chamber from which a catheter led to the skin surface. 2. The neutrophil granulocyte numbers and prostaglandin content of the wound fluid were studied, prostaglandins being identified by thin layer chromatography and measured by bio-assay. The major prostaglandin component was E2. 3. There was no clear relation between the amounts of prostaglandin present and the numbers of neutrophils in the fluid. 4. A subcutaneously implanted double chamber permitted perfusion of the lesion and thereby the study of the rate of prostaglandin production and the rate of disappearance of isotopically labelled prostaglandin from the lesion. The rate of leucocyte entry into the wound fluid could also be measured. 5. When the cutaneous muscle through which the chamber had been implanted was biopsied and incubated in vitro it was found to produce prostaglandin E2 in quantities which could be correlated with the weight of the incubated muscle. 6. These results suggest that a major contributor to the prostaglandin content of the wound fluid is the cutaneous striped muscle.

Animals

Local hyperalimentation of experimental granulation tissue.

The effect of local hyperalimentation on developing granulation tissue was studied in rats. Cylindrical hollow viscose cellulose sponge implants were used subcutaneously as an inductive matrix fro the growth of granulation tissue. In the first, control group the implants were kept untouched while the second, "sham" group was treated daily by withdrawing 1 ml of wound fluid from the central dead space of the implant and then injecting the fluid back. In the third, hyperalimentation group the aspirated wound fluid was substituted with a corresponding volume of sterile, nonpyrogenic solution containing a mixture of amino acids (Le-7402 A) and glucose, electrolytes and vitamins (Le-7402 B). Within the first week of tissue growth daily application of these nutritional substances caused a changeover of local tissue from predominantly anaerobic towards more oxidative metabolism. Measurement of nucleic acid and hydroxyproline contents indicated enhanced accumulation of cells and collagen in tissues receiving local hyperalimentation. The results combined with earlier data from our laboratory strongly suggest that several types of wounds, especially those containing a marked dead space or large regenerative area, exist in chronic lack of oxygen and other nutrients. Therefore, the healing process in these wounds can be stimulated, to a certain extent, by exposure to increased oxygen tension and/or by local hyperalimentation.

Amino Acids

High-Throughput and High-Sensitivity Biomarker Monitoring in Body Fluid by Fast LC SureQuant IS-Targeted Quantitation.

Targeted proteomics methods have been greatly improved and refined over the last decade and are becoming increasingly the method of choice in protein and peptide quantitative assays. Despite the tremendous progress, targeted proteomics assays still suffer from inadequate sensitivity for lower abundant proteins and throughput, especially in complex biological samples. These attributes are essential for establishing targeted proteomics methods at the forefront of clinical use. Here, we report an assay utilizing the SureQuant internal standard-triggered targeted method on a latest generation mass spectrometer coupled with an EvoSep One liquid chromatography platform, which displays high sensitivity and a high throughput of 100 samples per day. We demonstrate the robustness of this method by quantifying proteins spanning six orders of magnitude in human wound fluid exudates, a biological fluid that exhibits sample complexity and composition similar to plasma. Among the targets quantified were low-abundance proteins such at tumor necrosis factor A and interleukin 1-β, highlighting the value of this method in the quantification of trace amounts of invaluable biomarkers that were until recently hardly accessible by targeted proteomics methods. Taken together, this method extends the toolkit of targeted proteomics assays and will help to drive forward mass spectrometry-based proteomics biomarker quantification.

Humans

Mechanism of obstruction of closed-wound suction tubing.

The mechanism of obstruction of closed-wound drainage tubing was investigated by means of coagulation tests performed on wound drainage fluid and by examination of the contents of the tubes after their removal. Although clotting is commonly thought to be responsible for the obstruction, the wound drainage fluid was found to be essentially incoagulable and little fibrin was evident within the tubes. By contrast, bits of tissue were frequently found within the tubes, and these frequently virtually occluded the lumen. This observation, that tissue fragments are responsible for tube obstruction, permits a rational approach to the solution of this problem. For example, meticulous wound flushing and irrigation, or perhaps tubing of different design, might lead to a reduced incidence of tube failure.

Blood Vessels

Comparison between moist swab and tissue biopsy methods for quantitation of bacteria in experimental incisional wounds.

The relationship between tissue-associated and wound fluid-associated levels of bacteria in closed, healing experimental incisional wounds seeded with Escherichia coli was examined in 200 rats. Tissue specimens and moist swab specimens were taken simultaneously and cultured by a single plate serial dilution method. Colony counts of approximately 10(5) bacteria/gm of tissue were found to be equivalent to colony counts of 10(3) bacteria/ml of specimen obtained on a moist swab. Moist swab sampling of an incisional wound therefore offers a direct and simple method of ascertaining infection (defined by others as greater than 10(5) bacteria/gm of tissue). Moist swab sampling obviates the necessity of surgical manipulation of the patient and subsequent weighing and grinding of tissue in the laboratory.

Animals

Systematic bacteriological monitoring of intensive care unit patients: the results of a twelve month study.

The results from the bacteriological monitoring of 464 ITU patients are presented. The specimens analysed include urine, sputa, tracheostomy swabs, central venous pressure line catheter tips, blood cultures, wound drainage fluid, cerebro-spinal fluid, pleural fluid and peritoneal dialysate samples. Guidelines which may be used to differentiate between colonisation and infection and factors, notably those related to antibiotic usage, which are associated with increased risk of infection are described. Certain bacterial and fungal infections which are more peculiar to the compromised host are also discussed.

Bacterial Infections

Fungus infections after liver transplantation.

The problem of fungus infections after liver transplantation was studied. In 100 consecutive recipients of orthotopic liver homografts there were 10 and 8 examples, respectively, of localized and disseminated infections caused by Candida species. Candidemia was demonstrated in 8 of these 18 patients. One patient who had a localized Candida infection also had disseminated cryptococcosis. An additional 31 patients were infested in that Candida could be cultured from sites where it is not normally found, such as the blood (8 examples), urine (8), ascitic fluid (8), and wounds (22). This exorbitant incidence of monilial infections and infestations was associated with a high frequency of complications involving the homograft as well as the hosts' gastrointestinal tract during the post-transplantation period. The yeasts found in blood, urine, ascitic fluid and elsewhere were thought to have originated from the gut. Ten of the 100 patients had aspergillosis which was localized in 7 instances and disseminated in 3. The lung was the most frequently affected organ. The fungus infections played a contributory role in the downhill course of our patients but in the event of death more fundamental and more frequent causes of failure were technical complications involving the homografts, difficulties in controlling rejection with reasonable immunosuppressive doses and bacterial sepsis. Suggestions have been made for the better control of fungal infections in liver recipients.

Ascitic Fluid

Shortening of the bleeding time in thrombocytopenic rabbits after exposure of jugular vein to high aspirin concentration.

Aspirin, in doses which inhibit platelet thromboxane A2 production, prolongs the bleeding time but this effect on the bleeding time is lost when doses of aspirin which also inhibit vessel wall prostaglandin I2 (PGI2) production are used. PGI2 is both a potent inhibitor of platelet aggregation and a powerful vasodilator. We have investigated the contribution of the vascular effect of PGI2 on hemostasis by studying the effect of high concentrations of aspirin on the jugular vein bleeding time in severely thrombocytopenic rabbits and on the loss of non-platelet-containing fluid from standard puncture wounds in aspirin-treated veins perfused under constant pressure. After aspirin treatment, the bleeding time was significantly shortened in both normal and thrombocytopenic rabbits. This effect was associated with a decreased production of PGI2-like material by the vessel wall and a reduction in the volume of fluid lost from the standard puncture wound in the jugular vein. The effect of aspirin on the bleeding time and on PGI2 production was relatively short-lived and the bleeding time returned to normal within 2-3 hours. These observations indicate that PGI2 can influence hemostasis by mechanisms independent of platelet aggregation.

Animals

Preventing disruption of abdominal wounds.

The use of silver wire retention sutures requires added operating room time and expense, and it incurs the potential hazards of caught viscera, the possibility of more adhesions, leakage of intraperitoneal fluid through the exit wound, mild infection of exit wounds, and, on removal of the sutures, pain despite sedation, cutting of viscera, and a residual cross-hatched scar. Nevertheless, we believe that use of the technic is amply justified by the prevention of abdominal wound disruption, ease in visualization of the wound, ease in providing equal distribution of wound tension and periodic adjustment of it, and in reapproximation of superficial tissues in the event of wound breakdown, as well as in the elimination of need for any external supportive device. The existence of a voluminous literature on the placing, making, and closing of abdominal incisions suggests that no single ideal method exists. Major disruption of abdominal wounds continues to be a serious problem, despite the infrequency of its occurrence. The use of through-and-through semirigid silver wire retention sutures with adjustable tension as a complement to the routine layered closure can prevent disruption of abdominal wounds while causing only minor complications.

Abdominal Injuries

Reversed unidirectional transport in a Janus polyurethane/alginate dressing for directional postbiotic delivery to infected wounds.

Probiotic-derived postbiotics exhibit significant potential for infected wound control; however, their effective and localized delivery at wound sites remains a challenge. This study developed a polyurethane/alginate composite nonwoven via electrospinning to establish a postbiotic delivery platform for Bifidobacterium bifidum BD-1 (PU/Alg/BD-1). The beaded fibrous hydrophobic PU layer and hydrophilic Alg layer form a wettability gradient, enabling reversed unidirectional fluid transport toward the wound interface while inhibiting backflow. In vitro results showed that PU/Alg/BD-1 exhibited significant antibacterial activity against Staphylococcus aureus and Escherichia coli and good cytocompatibility with a hemolysis rate of <5%. Targeted metabolomic analysis revealed multiple organic acids in the BD-1 metabolites, which contributed to its antibacterial activity. In vivo microbial analysis verified that PU/Alg/BD-1 effectively reduced the relative abundance of Staphylococcus at the wound site while increasing the proportions of Corynebacterium and Psychrobacter. This microbial modulation contributed to infection control in a rat full-thickness infected wound model, accompanied by a shift in the macrophage phenotype and the downregulation of inflammatory factors including IL-6, TNF-&#x3b1;, and TGF-&#x3b2; in the PU/Alg/BD-1 group. Compared with the blank control, conventional gauze, PU/Alg, and BD-1 groups, PU/Alg/BD-1 significantly promoted wound contraction and re-epithelialization and enhanced collagen deposition. Hence, this study provides an effective material construction strategy for the application of probiotic-derived postbiotics to promote wound healing, demonstrates the potential of BD-1 to regulate the wound microenvironment and accelerate healing, and thereby offers a novel approach for the treatment of infected wounds.

Journal Article

Reaction of superficial bursae in response to specific disease stimuli.

Although microscopic studies have shown similarities between bursal and joint membranes, little is known about bursae and their response to disease states. Eighty-six cases of superficial bursitis due to trauma, bacterial infection, or gout were reviewed and compared with cases of joint inflammation due to the same etiologies. In traumatic bursitis the bursal fluid mucin test was more abnormal and the viscosity lower than that of joint fluid in traumatic arthritis. The bursal fluid total leukocyte count of septic bursitis was less than 20,000/mm3 in 8 of 13 cases but in only 1 of 21 synovial fluids from cases of septic arthritis (P = 0.005). In gouty bursitis the mean total leukocyte count of bursal fluid was 2800/mm3, compared with a mean synovial fluid total leukocyte count of 28,700 in gouty arthritis (P less than 0.02). These findings indicate that superficial bursae react less intensely than diarthrodial joints to specific disease stimuli and that a relatively low bursal fluid leukocyte count is often present in cases of septic and gouty bursitis.

Arthritis

The effect of intermittent pneumatic compression on the hand after fasciectomy.

Thirty-nine patients undergoing surgery for Dupuytren's contracture were included in a fully randomised, matched, prospective trial to evaluate the effect of intermittent compression on the post-operative hand. The study revealed a definite improvement in the treated, over the control patients. There was a distinct decrease in oedema with almost immediate return to normal hand function in the group treated by compression. Other benefits accrued from the earlier subsidence of pain in treated hands requiring reduced or no analgesia and from the earlier expression of fluid discharge from the wound thus preventing haematoma formation and the consequent ills.

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