[Robotic surgery]. / Robotsebészet.

Due to the fast spread of laparoscopic cholecystectomy, surgical procedures have been changed essentially. The new techniques applied for both abdominal and thoracic procedures provided the possibility for minimally invasive access with all its advantages. Robots - originally developed for industrial applications - were retrofitted for laparoscopic procedures. The currently prevailing robot-assisted surgery is ergonomically more advantageous for the surgeon, as well as for the patient through the more precise preparative activity thanks to the regained 3D vision. The gradual decrease of costs of robotic surgical systems and development of new generations of minimally invasive devices may lead to substantial changes in routine surgical procedures.

[New possibilities in practical education of surgery]. / Új lehetoségek a sebészet gyakorlati oktatásában.

The fast spread of laparoscopic surgery in the surgical community also required introduction of new methods of surgical education of these techniques. Training boxes applied for this reason meant a considerable help. The technique of the virtual reality introduced simulation, which is a new possibility in education. For the first time in the history of surgery we can measure medical students' or residents' dexterity and one can get acquainted with a surgical procedure in the form of "serious games". By application of the up-to-date imaging methods we can plan the movements of the surgeon's hand even before the planned operation, practice and repeating can contribute to the safety of the real procedure. Open surgical procedures can be practiced on plastic phantoms mimicking human anatomy and the use of interactive touch devices and e-learning can also contribute to practical education of surgery.

Long-term following-up of viability of spleen autotransplants in the Beagle canine model.

PURPOSE: To examine the possible late complications of splenectomy or spleen autotransplantation in large laboratory animal model, in which we need non-invasive or minimal-invasive methods for long-term monitoring of the experimental animals. METHODS: Experimental groups of beagle dogs were: non-operated control, sham-operated control, splenectomy, spleen autotransplantation with 5 or 10 spleen-chips taken into the greater omentum (Furka's technique). Prior to operations, on the 1(st) postoperative week, monthly till the 6(th) as well as in the 9(th) and 12(th) month, hemorheological examinations were performed. In postoperative 12(th) month colloid scintigraphy and diagnostic laparoscopy were carried out. At the end of the investigation comparative morphological examinations were performed, too. RESULTS: From the 4(th)-5(th) postoperative month filtration function of spleen-autotransplants showed particular restoration compared to splenectomy group. However, the functional results did not reach the values of the control or sham-operated groups. Sham-operated control's scintigraphy nicely showed activity in the spleen. In spleen autotransplantation-groups scintigraphy indicated well the activity of spleen-chips. During diagnostic laparoscopy spleen-chips with their blood supply were found. Histologically, the structure of spleen-autotransplants was similar to normal splenic tissue. CONCLUSIONS: The autotransplants are regenerated, their functions have been partly restored, and thus spleen autotransplantation may prevent the possible complications of splenectomy. These parameters and the presented investigative protocol are suitable for long-term following-up of viability of the spleen-autotransplants.

Long-term following-up of viability of spleen autotransplants in the Beagle canine model / Avaliação em longo prazo da viabilidade do autotransplante de baço em cães Beagle

PURPOSE: To examine the possible late complications of splenectomy or spleen autotransplantation in large laboratory animal model, in which we need non-invasive or minimal-invasive methods for long-term monitoring of the experimental animals. METHODS: Experimental groups of beagle dogs were: non-operated control, sham-operated control, splenectomy, spleen autotransplantation with 5 or 10 spleen-chips taken into the greater omentum (Furka's technique). Prior to operations, on the 1st postoperative week, monthly till the 6th as well as in the 9th and 12th month, hemorheological examinations were performed. In postoperative 12th month colloid scintigraphy and diagnostic laparoscopy were carried out. At the end of the investigation comparative morphological examinations were performed, too. RESULTS: From the 4th-5th postoperative month filtration function of spleen-autotransplants showed particular restoration compared to splenectomy group. However, the functional results did not reach the values of the control or sham-operated groups. Sham-operated control's scintigraphy nicely showed activity in the spleen. In spleen autotransplantation-groups scintigraphy indicated well the activity of spleen-chips. During diagnostic laparoscopy spleen-chips with their blood supply were found. Histologically, the structure of spleen­autotransplants was similar to normal splenic tissue. CONCLUSIONS: The autotransplants are regenerated, their functions have been partly restored, and thus spleen autotransplantation may prevent the possible complications of splenectomy. These parameters and the presented investigative protocol are suitable for long-term following-up of viability of the spleen-autotransplants.

OBJETIVO: Examinar as possíveis complicações tardias da esplenectomia ou do autotransplante de baço em modelo animal de grande porte, no qual faz-se necessário o uso de métodos não invasivos ou minimamente invasivos para monitorizar os animais de experimentação. MÉTODOS: Grupos experimentais de cães beagle foram: não-operados controle, sham-operados controle, esplenectomia, autotransplante de baço com 5 ou 10 fatias de baço colocados no grande omento (técnica de Furka). Antes das operações, na 1ª semana de pós­operatório, mensalmente até 6o.assim como no 9º. e 12º. mês, foram realizados exames hemorreológicos. No 12º. mês de pós-operatório, cintilografia colóide e laparoscopia diagnóstica foram realizadas. Ao final do experimento, exames morfológicos comparativos foram realizados também. RESULTADOS: A partir do 4º-5º mês pós-operatório, a função de filtração dos baços autotransplantados mostraram particular restauração comparados ao grupo esplenectomia. Entretanto, os resultados funcionais não alcançaram os valores dos grupos controle ou sham-operados. A cintilografia dos controles sham-operados mostraram atividade no baço. Nos grupos de autotransplante, a cintilografia indicou bem a atividade das fatias de baço. Durante a laparoscopia diagnóstica, as fatias de baço com seu suprimento sanguíneo foram encontrados. Histologicamente, a estrutura dos autotransplantes de baço foi similar ao tecido normal de baço. CONCLUSÕES: Os autotransplantes são regenerados, suas funções foram parcialmente restauradas, e então ao autotransplantate esplênico pode prevenir as possíveis complicações da esplenectomia. Estes parâmetros e o protocolo experimental são adequados para o seguimento em longo prazo da viabilidade de autotransplantes esplênicos.

Hemorheological consequences of hind limb ischemia-reperfusion differs in normal and gonadectomized male and female rats.

It is known that hemorheological parameters show gender differences that might be altered by gonadectomy (GoE). Since micro-rheological parameters (erythrocyte deformability and aggregation) sensitively change during and after ischemia-reperfusion (I/R), the question arises whether the hemorheological effects of I/R may show gender differences and further changes might be expected when GoE and I/R are additive. Sprague-Dawley rats were divided into six groups: Control males and females, I/R males and females with 1-hour hind limb ischemia, GoE + I//R males and females when 3 months after bilateral gonadectomy the I/R was induced. Before and just after ischemia, and on the 1st-3rd-5th-7th postoperative days blood samples were taken (lateral tail vein, 0.3-0.5 mL) for analyzing hematological parameters, erythrocyte's deformability (slit-flow ektacytometer) and aggregation (light-transmission aggregometer). Leukocyte and platelet counts raised markedly in gonadectomized animals during the investigated days. Hemorheological changes of I/R showed gender differences: significant impairment of erythrocyte deformability was found on the 1st-3rd postoperative days, expressed mostly in females. In gonadectomized females the postischemic deformability values were impaired. Erythrocyte aggregation index significantly raised by the 1st postoperative day, dominantly in males. It is suggested that gonadectomy may act as an additional rheological 'risk factor' related to blood micro-rheological parameters in ischemia-reperfusion.

Efeitos do alopurinol e precondicionamento na apoptose devido a isquemia-reperfusão em duplo segmento de jejuno em cães / Effects of allopurinol and preconditioning on apoptosis due to ischemia-reperfusion on a double jejunum-segment canine model

PURPOSE: To investigate the duration of apoptosis caused by ischemia-reperfusion in the intestine in a new double jejunum-segment model, and to analyze the protective effects of allopurinol or ischemic preconditioning (IPC). METHODS: In Experiment I for harvesting the double jejunum-segment model after laparotomy a 30-cm-long jejunum part was selected on mongrel dogs (n=24). End-to-end anastomoses were performed at both ends and in the middle of the jejunum part, creating two equal segments. In one segment ischemia was induced by occluding the supplying vessels, the other segment served as control. Tissue samples for detecting apoptosis were taken at 30th minutes, 1st, 2nd, 4th, 6th, 8th, 12th and 24th hours of reperfusion. In Experiment II using the same model the 4-hour reperfusion time period, allopurinol (50 mg/kg) pre-treated and IPC (3 cycles of 5x1) groups (n=5 per each) were also investigated. RESULTS: In Experiment I the greatest apoptotic activity was detected at the 4th and 6th hour of reperfusion (14.2 ± 1.31 and 16.3 ± 1.05 per visual field at 40x magnification). In Experiment II Using the 4-hour reperfusion time period allopurinol pre-treatment increased the apoptotic activity (10.72 ± 0.47 per 50 intestinal villi) approximately two-fold than the IPC (6.72 ± 0.46 per 50 intestinal villi) did (p<0.05). CONCLUSIONS: Apoptotic activity has a characteristic time curve, reaching the highest values between the 4th and 6th hours after 30-minute intestinal ischemia. Ischemic preconditioning seemed to be protective against the morphological changes caused by intestinal ischemia-reperfusion.

OBJETIVO: Investigar a duração da apoptose causada pela isquemia-reperfusão no intestino em um novo modelo de duplo segmento de jejuno e analisar os efeitos protetores do alopurinol ou precondicionamento isquêmico (IPC). MÉTODOS: No experimento I para obter o modelo do duplo segmento de jejuno, após a laparotomia, uma parte de 30cm de comprimento de jejuno foi selecionada em cães mestiços (n=24). Anatomoses T-T foram realizadas em ambas as extremidades no meio do segmento de jejuno, criando dois segmentos iguais. Em um segmento foi induzida isquemia por oclusão dos vasos que o irrigavam e o outro segmento foi usado como controle. Amostras de tecido para detecção da apoptose foram obtidos aos 30 minutos, 1h, 2h, 4h, 6h, 8h, 12h e 24 horas de reperfusão. No experimento II usando o mesmo modelo, no tempo de reperfusão de 4 horas, foram investigados dois outros grupos (n=5 cada) usando precondicionamento com alopurinol (50 mg/kg) e IPC (3 ciclos de 5x1). RESULTADOS: No experimento I a maior atividade de apoptose detectada foi às 4h e 6h de reperfusão (14,2 ± 1,31 e 16,3 ± 1,05 no campo visual de 40x). No experimento II usando o período de 4horas de reperfusão o pré-tratamento com alopurinol aumentou a atividade apoptótica (10,72 ± 0,47) aproximadamente 2 vezes mais do que o IPC (6,72 ± 0,46) (p<0,05). CONCLUSÕES: A atividade de apoptose tem uma curva caractetística, atingindo maiores valores entre a 4ª e a 6ª horas após 30 minutos de isquemia intestinal. O precondicionamento isquêmico parece proteger contra alterações morfológicas causadas pela isquemia-reperfusão intestinal.

Effects of allopurinol and preconditioning on apoptosis due to ischemia-reperfusion on a double jejunum-segment canine model.

PURPOSE: To investigate the duration of apoptosis caused by ischemia-reperfusion in the intestine in a new double jejunum-segment model, and to analyze the protective effects of allopurinol or ischemic preconditioning (IPC). METHODS: In Experiment I for harvesting the double jejunum-segment model after laparotomy a 30-cm-long jejunum part was selected on mongrel dogs (n=24). End-to-end anastomoses were performed at both ends and in the middle of the jejunum part, creating two equal segments. In one segment ischemia was induced by occluding the supplying vessels, the other segment served as control. Tissue samples for detecting apoptosis were taken at 30th minutes, 1st, 2nd, 4th, 6th, 8th, 12th and 24th hours of reperfusion. In Experiment II using the same model the 4-hour reperfusion time period, allopurinol (50 mg/kg) pre-treated and IPC (3 cycles of 5x1) groups (n=5 per each) were also investigated. RESULTS: In Experiment I the greatest apoptotic activity was detected at the 4th and 6th hour of reperfusion (14.2 ± 1.31 and 16.3 ± 1.05 per visual field at 40x magnification). In Experiment II Using the 4-hour reperfusion time period allopurinol pre-treatment increased the apoptotic activity (10.72 ± 0.47 per 50 intestinal villi) approximately two-fold than the IPC (6.72 ± 0.46 per 50 intestinal villi) did (p<0.05). CONCLUSIONS: Apoptotic activity has a characteristic time curve, reaching the highest values between the 4th and 6th hours after 30-minute intestinal ischemia. Ischemic preconditioning seemed to be protective against the morphological changes caused by intestinal ischemia-reperfusion.

Morphological, hemodynamical and hemorheological changes of mature artificial saphenous arterio-venous shunts in the rat model.

Artificial femoral arterio-venous (AV) shunts are widely used in rodent models for studying shunt maturation and to optimize various surgical techniques. However, little is known about complex circulatory, microcirculatory, and hemorheological effects of end-to-side saphenous AV shunts. We aimed to study these parameters in mature AV shunts. Studying these questions in CD rats, end-to-side anastomoses were made between the left saphenous artery and vein. On the right-side the nonoperated saphenous vessels served as own control. Furthermore healthy control animals were also investigated. On the 8th to 12th postoperative week microcirculatory and blood flow measurements were performed and blood samples were taken both from the shunt's arterial and venous limbs and from the nonoperated side vessels. Hematological parameters, erythrocyte aggregation, and deformability were determined. The entire shunt and the control vessels were removed for histological examinations. The skin microcirculation on shunt side slightly increased on thigh and decreased on paws versus the nonoperated side. Blood flow measurements made directly on the vessels showed that arterial to venous blood flow rate ratio was 1.59 ± 0.29 on nonoperated side and 1.2 ± 0.13 on the shunt side, and 1.49 ± 0.05 in control animals. Erythrocyte aggregation and deformability worsened on the shunt side. Histologically increased number of smooth muscle elements and connective tissue were found in venous limb of the shunts. The artificial AV shunt between the saphenous artery and vein seems to be a suitable model for further functional-morphological and hemorheological examinations of hemodialysis in various states and diseases.

Examination of aggregation of various red blood cell populations can be informative in comparison of splenectomy and spleen autotransplantation in animal experiments.

Using "Furka's spleen-chip" autotransplantation technique, in a follow-up study we investigated erythrocyte aggregation and its influencing factors regarding to asplenic-hyposplenic conditions. Healthy beagle dogs were equally divided into 4 groups: "Sham"- sham operated control, "SE"-splenectomy, "AU-5" and "AU-10"-autotransplantation with 5 or 10 spleen chips by Furka-method after splenectomy. Blood sampling for current investigation occurred on the 12th, 18th, 20th and 24th postoperative months, besides comprehensive structural and functional examinations. In this study we focused on the aggregation properties of erythrocytes in native samples and of density separated "young" versus "old" erythrocytes (Myrenne MA-1 aggregometer). Fibrinogen concentration (Sysmex CA-500 coagulometer) and erythrocyte sedimentation rate (ESR) were also determined. SE group showed significantly elevated ratio of "old" and "young" erythrocyte aggregation index values on the 20th and 24th postoperative months (1.5 and 2.16, respectively) comparing to the AU (1.14 and 1.05) and Sham groups (1.2 and 1.27). In SE group fibrinogen concentration was mildly higher and ESR almost doubled versus control. The greater differences experienced between "young" and "old" erythrocytes may reflect decreased splenic filtration function. Consequently, the comparison of aggregation and deformability properties of "young" and "old" erythrocytes may give important information about the function of the spleen.

Splenic function and red blood cell deformability: The beneficial effects of spleen autotransplantation in animal experiments.

Erythrocyte deformability could be a suitable parameter reflecting hyposplenic-asplenic conditions after splenectomy or spleen autotransplantation. The aim of this study was to analyze the efficacy of spleen autotransplantation using filtrometry and ektacytometry for testing erythrocyte deformability, besides complex structural, functional, hematological, hemostaseological and immunological examinations. Healthy beagle dogs were subjected to the study in the following experimental groups: "Sham"-sham operated control, "SE"-splenectomy, "AU-5" and "AU-10"-autotransplantation with 5 or 10 spleen chips using "Furka's spleen chip" technique after splenectomy. Blood samples were taken before the operations and monthly afterwards for 1 year and bi-monthly till the end of the 2nd postoperative year. Erythrocyte deformability was measured by bulk filtrometry (relative cell transit time, RCTT) and on the 20th-24th postoperative months by ektacytometry (elongation index, EI), too. RCTT was elevated in operated groups till the 12th month. On the 20th/24th months "SE"-group expressed the highest level, while the "AU"-groups were close to "Sham"-group. Using ektacytometer we found that on the 20th/24th month both EImax and SS1/2 showed the lowest values (because of unusual curve characteristic) in "SE"-group and "AU"-groups almost reached the "SH"-group. Spleen autotransplantation partially restored the normal rheological conditions. Both filtrometry and ektacytometry was suitable to investigate hyposplenic-asplenic conditions.

Application of leukocyte antisedimentation rate calculation in investigation of spleen salvaging experimental surgical techniques.

Clinical studied showed leukocyte antisedimentation rate (LAR) changes in various conditions that modulate the immune system. There is a lack of LAR data in animal experiments concerning splenic function after splenectomy or spleen preserving operations, thus we aimed to include LAR in our ongoing inbred canine study on following-up spleen salvaging surgical techniques. Blood samples of healthy beagle dogs served as normal control. The experimental groups were: "SH"-sham operated, "SE"-splenectomy, "R1/3" and "R2/3"-one-third/two-third of the spleen resected, "AU-5" and "AU-10"-autotransplantation with 5/10 spleen chips using Furka's techniques. On the 12th postoperative month Rabigen-Mono+Vanguard-Plus-5 vaccines were given to the animals, which provoked immune response. Blood samples were taken before and one week after the vaccination. LAR was calculated by Bogar's method. In normal control group LAR was 3.11, leukocyte count was 12.04 G/l. In the experimental groups LAR varied between 2.4-7.3 before vaccination. After vaccination LAR increased in all groups. Although "SE" group expressed the largest changes (LAR = 16.56; 135%), the leukocyte count increased only by 12%. In "AU" groups the increase was smaller, while "R" groups showed slight changes only. LAR can be applicable in animal experiments and may serve as a supplementary parameter in hemorheological investigations of hyposplenic-asplenic states.

Hemorheological, morphological, and oxidative changes during ischemia-reperfusion of latissimus dorsi muscle flaps in a canine model.

Although ischemia-reperfusion (I/R) strongly influences muscle flap survival in reconstructive surgery, there is limited knowledge about its relation to hemorheological parameters and oxidative stress markers in flaps. In the present study we investigated these changes during I/R of latissimus dorsi muscle (LDM) flaps in beagle dogs. In four animals LDM flaps were prepared bilaterally. The right side served as control, while the left side's vascular pedicle was clamped for 60 minutes, and a 60-minute reperfusion was allowed afterward. Blood samples (0.5 ml each) were taken from the pedicle's vein bilaterally before and after the ischemia, and at the 5th, 15th, 30th, 45th, and 60th minutes of the reperfusion, for hematological and erythrocyte aggregation tests. In muscle biopsies, taken before and after I/R, histological investigations and tests for measuring gluthation-peroxidase (GSH-PX) activity, glutathione (GSH) and carbonyl concentrations, and thiobarbituric acid reactive substances (TBARS) content were carried out. In I/R side leukocyte count increased during the reperfusion with a peak at the 30th minute. Hematocrit continuously increased from the 15th minute. In the first 5 minutes of the reperfusion, erythrocyte aggregation increased, than tented to be normalized. In muscle homogenates GSH-PX activity did not change markedly, GSH content slightly decreased, carbonyl and TBARS content increased during reperfusion. A 1-hour ischemia and reperfusion of LDM flaps caused local changes of leukocyte distribution and erythrocyte aggregation, supposedly due to the metabolic and inflammatory reactions. Oxidative damage during reperfusion was also demonstrated.

Changes of local and systemic hemorheological properties in intestinal ischemia-reperfusion injury in the rat model.

The aim of this study was to investigate intestinal ischemia-reperfusion and its local and systemic hemorheological relations in the rat. Ten anaesthetized female CD outbred rats were equally divided into 2 experimental groups. (1) Ischemia-reperfusion (I/R): the superior mesenterial artery was clipped for 30 minutes. After removing the clip, 60 minutes of the reperfusion was observed before extermination. Blood samples were taken from the caudal caval vein and from the portal vein before ischemia, 1 minute before and after clip removal, and at the 15th, 30th, and 60th minutes of the reperfusion. (2) Sham operation: median laparotomy and blood sampling were done according to the timing as in I/R group. Hematological parameters, red blood cell aggregation, and deformability were determined. Leukocyte count and mean volume of erythrocytes increased slightly but continuously in portal venous samples during the reperfusion period. Red blood cell aggregation values were higher in portal blood by the end of ischemia, and then became elevated further comparing to the caval venous blood. Both in caval and portal venous samples of I/R group red blood cell deformability significantly worsened during the experimental period compared to its base and Sham group. In portal blood red blood cell deformability was impaired more than in caval vein samples. Histology showed denuded villi, dilated capillaries, and the inflammatory cells were increased after a 30 minutes ischemia. In conclusion, intestinal ischemia-reperfusion causes changes in erythrocyte deformability and aggregation, showing local versus systemic differences in venous blood during the first hour of reperfusion.

Spleen autotransplantation. Morphological and functional follow-up after spleen autotransplantation in mice: a research summary.

In 1986, we started the research on spleen surgery aimed at saving the splenic mass after its traumatic injury, with elaboration of special resection and autotransplantation techniques. The researches started on mongrel dogs and were continued on inbred mice and beagle dogs with complex histological, imaging, and laboratory investigations, following-up the function and the regeneration of autotransplanted spleen chips. Performing research on mice provided more immunological methods, such as lymphocyte subsets, immunoglobulin levels, and monitoring the phagocytic functions. Researches showed evidence also on the presence of apoptosis, furthermore, stem cell studies on regeneration and functional restoration of the spleen chips are in progress. Our results contributed to two multidisciplinary guidelines in Hungary: (1) One of them is under preparation and underlines the importance of spleen saving methods after traumatic splenic injury; (2) The second guideline shows that hemorheological changes can be early indicators of the increased sensitivity to postsplenectomy infections.

Functional sphincter ani externus reconstruction for treatment of fecal stress incontinence using free latissimus dorsi muscle transfer with coaptation to the pudendal nerve: preliminary experimental study in dogs.

The external anal sphincter (EAS) is a skeletal muscle capable of voluntary contraction to prevent accidental defecation. Current reconstructive options for a severely damaged EAS using local muscle flaps are not always adequate for functional repair. The present preliminary experimental model was designed to assess the feasibility of a neuromicrovascular latissimus dorsi muscle transfer for functional external spincter muscle reconstruction. In nine mongrel dogs, the anal sphincter muscles were totally resected, leaving a mucosal canal in place. A segmental latissimus dorsi muscle was shaped around the anal canal in a circular fashion, with coaptation to the pudendal nerve, and vessel anastomosis at the ischiorectal fossa. Functional evaluation was performed using electromyogram, sphincter manometry, video documentation, and histologic examination with standard and immunohistochemical staining. After 8 months, the remaining three eligible dogs were continent. Muscle function was verified by means of electromyogram, sphincter manometry, and a video record. Histologic and immunohistochemical examination confirmed the functional results, showing only minor zones of fatty and fibrous degeneration. Transplantation of a segmental latissimus dorsi muscle with vascular anastomosis and coaptation to the pudendal nerve has proved to be successful in restoring (voluntary) anal continence experimentally in dogs. Its feasibility for perfect orientation as a neosphincter seems to be superior to any pedicled muscle flap. However, these preliminary results deserve further investigation prior to considering application in humans.

[Conceptions about microsurgical education. What were 5,460 hours of microsurgical basic education enough for?]. / Gondolatok a mikrosebészeti oktatásról. Mire volt elég az 5460 óra a mikrosebészeti alapoktatásban?

Microsurgical education has an involuting role in the respects both of the effective clinical and successful research work. In Hungarian literature little number of paper can be found that deal with microsurgical educational methods, their results and efficacy. In our department for several decades we have taught the basics of microsurgery at different education levels (graduate and postgraduate), giving the possibility for practicing and developing as well. In this paper we summarize our educational objective and subjective experiences of more than 10 years and 5460 education hours in microsurgical basic courses. Besides the numeric summaries it became clear that the teacher's or tutor's and the candidates' experience and their ability to attain the basics of microsurgery are very important. According to our experience the most effective method is the education in small groups: one tutor and 2-3 candidates can reach the final goal, the basic microsurgical skill.

Cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted mice.

BACKGROUND: Our department worked out a modified surgical form of spleen autotransplantation earlier, named "spleen apron method" introduced already into the clinical practice. Recently we tested the immunological changes in a group of patients autotransplanted with about 10-15% of their spleen, what was the at least always implantable amount after the severe splenic injuries. In the current work we aimed at measuring some cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted inbred mice two and eight months after the operations in order to get more unambiguous results than that we could obtain in our patients with this technique. MATERIALS AND METHODS: We divided 96 two months old Balb/c female mice into eight groups (n = 12/group). The group of controls, sham operated, splenectomized and autotransplanted animals with two and eight months of survival time after the operations. During the autotransplantation we inserted the same amount of spleen, five slices, "chips," about 10-15% of total mass of spleen, into the greater omentum similarly as it was used in the patients. The concentration of serum proteins were measured by laser nephelometry. The lymphocyte subsets were analyzed by flow cytometry. RESULTS: We found that two months after the operations the number of CD 19+ B-cells increased in the splenectomized but decreased in the autotransplanted animals. Eight months after the operations the number of both CD3+ T and CD19+ B lymphocytes decreased both in the splenectomized and autotransplanted animals compared to the controls and sham operated mice. However, the numbers of T and B cells were slightly but not significantly higher in the autotransplanted than in the splenectomized mice. The serum level of IgM was also decreased in the splenectomized and autotransplanted mice at both time points, however, eight months after the operations the concentration of IgM was significantly higher in the autotransplanted group than in the splenectomized animals. CONCLUSION: The effects of autotransplanted "chips" were different at the various ages of the animals. Additionally, they showed some immunological benefit being quantitatively in accordance to the amount of the transplanted spleen. The elevated level of serum IgM what we found in the autotransplanted mice even with this amount of transplanted spleen eight months after the operations, however, might have the potentially greatest importance compared to splenectomy. These experiments can prove that the attempts for autotransplantation may have real perspectives but their efficacy depends on the amount of the successfully transplanted (saved) mass of spleen.

Learning microsurgical suturing and knotting techniques: comparative data.

Correctly performed surgical sutures are the basis of surgical safety. This retrospective survey was conducted among participants (n = 263) taking graduate and postgraduate courses between 2000-2004. Placement of sutures, time to perform knotting, safety of knots, and quality of knot stability were tested. None of the students had been previously instructed in microsurgical techniques. At the beginning of the training program, 90-95% of participants damaged the suturing thread at several places. By the end of the course, knotting times significantly decreased in both groups. Graduates decreased their time from 6.8 +/- 2.34 min to 3.28 +/- 0.71 min (mean +/- standard error of the mean), and postgraduates decreased their time from 5.02 +/- 3.25 min to 1.54 +/- 0.54 min (mean +/- standard error of the mean). In our opinion, "mass training" to teach the basics of microsurgery is not a good approach. Instead, individual training should be provided, as tutors offer invaluable advice, and adjust almost each stitch and knot during teaching sessions.

Distribution of peripheral blood cells in mice after splenectomy or autotransplantation.

Our aim was to compare the distribution changes of peripheral leukocytes and erythrocytes in splenectomized and spleen-autotransplanted BALB/c female mice (n = 96), 2 and 8 months after surgery. In total, there were eight groups of animals: splenectomy, autotransplantation, sham, and untreated controls at both time points. We used the spleen-apron method of Furka et al. (Khirurgiia (Mosk) 1989;9:125-127), inserting five spleen chips into the greater omentum, for autotransplantation. Quantitative and qualitative blood cell counts and the phagocytic activity of cells (stimulated with zymosan) were determined. In splenectomized animals, the number of neutrophils significantly increased 8 months after surgery. The greatest phagocytic activity of neutrophils, however, was observed in autotransplanted animals of the same age. In splenectomized animals, erythrocyte volumes were significantly higher in the second postoperative month, but normalized by the eighth month. In conclusion, spleen autotransplantation has some beneficial effects, including clearing erythrocytes and preserving the phagocytic activity of neutrophils in peripheral blood.

Hemorheological follow-up after splenectomy and spleen autotransplantation in mice.

We previously reported on a spleen autotransplantation model in mice, with spleen function studies at 6 weeks. The present study was undertaken to investigate long-term hemorheological functions. A/J and BALB/c inbred mice were divided into four groups: control, sham surgery (SH), splenectomy (SE), and spleen autotransplantation (AU). Hematological and hemorheological parameters were determined. Leukocyte counts in the SE and AU groups were significantly higher than in controls, while hematocrit levels were markedly lower. Mean erythrocyte volume did not change significantly. Platelet counts in the AU group were significantly lower, and red blood cell deformability was significantly worse in the SE group. The AU group also had increased cell transit time, but it was less than that in the SE group. The SE group showed the highest fibrinogen levels. We conclude that there are some consistent differences in hematological parameters between splenectomy and spleen autotransplantation. These data suggest that spleen autotransplantation may partially restore hemorheological functions following splenectomy.