Changes in microcirculation of small intestine end-to-end anastomoses in an experimental model.

INTRODUCTION: Sufficient perfusion is essential for a safe intestinal anastomosis. Impaired microcirculation may lead to increased bacterial translocation and anastomosis insufficiency. Thus, it is important to estimate well the optimal distance of the anastomosis line from the last mesenterial vessel. However, it is still empiric. In this experiment the aim was to investigate the intestinal microcirculation at various distances from the anastomosis in a pig model. MATERIALS AND METHODS: On 8 anesthetized pigs paramedian laparotomy and end-to-end jejuno-jejunostomy were performed. Using Cytocam-IDF camera, microcirculatory recordings were taken before surgery at the planned suture line, and 1 to 3 mesenterial vessel mural trunk distance from it, and at the same sites 15 and 120 min after anastomosis completion. After the microcirculation monitoring, anastomosed and intact bowel segments were removed to test tensile strength. RESULTS: The proportion and the density of the perfused vessels decreased significantly after anastomosis completion. The perfusion rate increased gradually distal from the anastomosis, and after 120 min these values seemed to be normalized. Anastomosed bowels had significantly lower maximal tensile strength and higher slope of tensile strength curves than intact controls. CONCLUSION: Alterations in microcirculation and tensile strength were observed. After completing the anastomosis, the improvement in perfusion increased gradually away from the wound edge. The IDF device was useful to monitor intestinal microcirculation providing data to estimate better the optimal distance of the anastomosis from the last order mesenteric vessel.

Heparin-like effect of a dual antiplatelet and anticoagulant (APAC) agent on red blood cell deformability and aggregation in an experimental model.

Treatments with different antithrombotic agents can affect micro-rheological variables, such as red blood cell (RBC) deformability and aggregation. Since the effect of dual antiplatelet and anticoagulant (APAC) treatment on micro-rheology is unknown, we aimed to investigate the effect of different intravenous doses of APAC on hematological and micro-rheological variables in a porcine model. Two groups were formed (APAC group, Control group), and blood was collected from the animals at preset intervals. Hematological variables, RBC deformability, and aggregation were measured. We observed an improvement in the RBC deformability measured at a low shear stress range (< 3 Pa). However, after both doses, a decrease in the maximal elongation index of RBC values occurred in the APAC group. RBC aggregation increased after APAC bolus dose, while it gradually and dose-dependently decreased. Supposedly, the improvement in RBC deformability that was observed at a lower shear rate could facilitate aggregation. Administration of APAC and unfractionated heparin (UFH) caused comparable changes in hematological and hemorheological variables. Signs of thrombosis or bleeding did not occur. APAC and UFH had comparable micro-rheological effects.

Lernaea cf. cyprinacea infection in agile frog (Rana dalmatina) tadpoles in Hungary.

The ectoparasitic copepod Lernaea cyprinacea (anchor worm) has more than 100 host species among teleost fishes and affects cyprinids both in fish farms and natural waters. In addition, while L. cyprinacea infection in amphibians has been recorded in Asia, North and South America, there is no data available in the literature on their presence in Europe. In this study, we first reported L. cf. cyprinacea parasitising an anuran tadpole in Europe. Specimens of L. cf. cyprinacea were observed attached to a tadpole of the agile frog (Rana dalmatina), which was caught during fishing of crucian carp fingerlings (Carassius carassius) from a small fish pond in Hungary during the summer of 2012. The infected tadpole was collected from a rearing pond, where juvenile crucian carps were kept. The tadpole was inspected in the laboratory, and digital photos were taken. The parasites were found attached to the body-tail junction and to the leg of the tadpole (at Gosner stage 41). The parasite species was identified as L. cf. cyprinacea based on morphological traits.

Application of a Self-developed, Low-budget Indocyanine Green Camera in Surgical Imaging - a Single Institution's Experiences.

INTRODUCTION: Indocyanine green is a fluorescent dye, the use of which is becoming more and more widespread in different areas of surgery. Several international studies deal with the dye's usefulness in intraoperative angiography, the localization of tumors, the more precise identification of anatomical structures, the detection of lymph nodes and lymph ducts, etc. The application of the dye is safe, but a suitable equipment park is required for its use, which entails relatively high costs. OBJECTIVES: The aim of our research is to create a detector system on a low budget, to be used safely in everyday practice and to illustrate its operation with practical examples at our own institute. METHODS: By modifying a web camera, using filter lenses and special LEDs, we created a device suitable for exciting and detecting indocyanine green fluorescence. We prove its excellent versatility during the following procedures at our institute: breast tumor surgery, kidney transplantation, bowel resection, parathyroid surgery and liver tumor resection. RESULTS: The finished camera has an LED light source with a peak wavelength of 780 nm, and the incoming light is filtered by a bandpass filter with a center wavelength of 832 nm. A low budget ($112), easy-to-use tool was created, which is suitable for taking advantage of the opportunities provided by indocyanine green.

Molecular evidence of the absence of Metagonimus yokogawai (Katsurada, 1912) in Europe: report of Metagonimus sp. in cyprinoid fish from the River Danube in Hungary.

Trematodes of the genus Metagonimus Katsurada, 1912 (Digenea: Heterophyidae) are zoonotic parasites that cause infections in humans, with most cases reported in Southeast Asia. Larvae from the second intermediate host, called metacercariae, of one of human-infecting species, M. yokogawai (Katsurada, 1912), have been reported from cyprinoid fish in Europe. In the present study, we provided DNA-based evidence that metacercariae of Metagonimus, which are commonly found in the scales of various cyprinoids in Central Europe (Danube River in Hungary) do not belong to M. yokogawai. Sequence analysis of the ITS region, 28S rDNA, and cox1 genes showed that this species is clearly distinct from all Asian species, including M. yokogawai, which probably does not occur in Europe. Metacercariae from cyprinoids might belong to Metagonimus romanicus (Ciurea, 1915), an insufficiently known species described from Romania.

Impact Assessment of Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) and Hemostatic Sponge on Vascular Anastomosis Regeneration in Rats.

The proper regeneration of vessel anastomoses in microvascular surgery is crucial for surgical safety. Pituitary adenylate cyclase-activating polypeptide (PACAP) can aid healing by decreasing inflammation, apoptosis and oxidative stress. In addition to hematological and hemorheological tests, we examined the biomechanical and histological features of vascular anastomoses with or without PACAP addition and/or using a hemostatic sponge (HS). End-to-end anastomoses were established on the right femoral arteries of rats. On the 21st postoperative day, femoral arteries were surgically removed for evaluation of tensile strength and for histological and molecular biological examination. Effects of PACAP were also investigated in tissue culture in vitro to avoid the effects of PACAP degrading enzymes. Surgical trauma and PACAP absorption altered laboratory parameters; most notably, the erythrocyte deformability decreased. Arterial wall thickness showed a reduction in the presence of HS, which was compensated by PACAP in both the tunica media and adventitia in vivo. The administration of PACAP elevated these parameters in vitro. In conclusion, the application of the neuropeptide augmented elastin expression while HS reduced it, but no significant alterations were detected in collagen type I expression. Elasticity and tensile strength increased in the PACAP group, while it decreased in the HS decreased. Their combined use was beneficial for vascular regeneration.

Extreme pathological symptom generated by Dermocystidium koi infection of common carp (Cyprinus carpio L.) in Hungary.

Dermocystidiosis in common carp (Cyprinus carpio L.) as a skin infection in the form of small, pea-sized bulging nodules is well-known in Hungary. Its occurrence is sporadic, and the pathogenic effect is generally moderate. Nonetheless, here we report an unusual case of the infection when a mass of dermocystid spore-filled hyphae-like cysts formed a large tumour-like nodule under the skin intruding deeply in the red muscle on one side occipital region of the fish body. Histological sections showed that the tumour-like nodule was composed of two layers, a pale peripheral part with a high mass of cysts surrounded by some cell debris of muscle cells and a dark central part with a less dense meshwork of tubular cysts which were embedded in the partially damaged red muscle tissue. No infection was found in the white muscle. Sequences of 18S rDNA of spores from the tumour-like nodule showed a complete identity to sequences from typical bulging nodules in skin. They were also highly similar to the sequences of D. anguillae clones (99.5%-99.4%), to some samples of D. salmonis (99.7% and 99.4%), and to the Dermocystidium sp. CM-2002 (99.4%) from European perch.

Mycoplasma infections in freshwater carnivorous fishes in Hungary.

The abundance of mycoplasma-like microorganisms seems to be common in the intestinal tract of fish, but their occurrence in other organs can be considered pathological. Isolation of piscine mycoplasmas is essential to prove their virulence. In this study, a total of 100 specimens of nine freshwater fish species collected from natural waters and fish farms were analysed for the prevalence of mycoplasmas in different extraintestinal organs. Our results demonstrate that mycoplasmas are present in carnivorous fish (pike, wels catfish, black bullhead and pike perch) than in cyprinids more frequently. Among the surveyed organs, they colonize mainly the gills, the kidney and the livers. Although mycoplasmas could be detected in one-fifth of samples by molecular assays, the isolation of mycoplasmas belonged to the M. sualvi cluster within the hominis group of Mycoplasmataceae, but was separate from M. mobile, has been successful only from some specimens originated from wels catfish that was feasible only by special preparation steps (prefreezing, filter sterilization and incubation conditions (long culturing period, etc.). The low-efficiency of isolation trials indicates an extensive gap in our knowledge about piscine mycoplasmas, especially in the field of their nutritional requirements.

Molecular detection of a novel cyprinid herpesvirus in roach (Rutilus rutilus) and asp (Leuciscus aspius) showing typical signs of carp pox disease.

In the early spring of 2018, in Lake Balaton (Hungary), a roach (Rutilus rutilus) and an asp (Leuciscus aspius) were found in an fish trap at the outlet of the river Sió showing typical signs of the so-called carp pox disease, such as foci of epidermal hyperplasia on the head and the whole body surface, including the fins. Molecular tests revealed the presence of the DNA of an unknown fish herpesvirus. Three genes encoding the DNA-dependent DNA polymerase, major capsid protein and ATPase subunit of terminase were amplified and sequenced from the alloherpesviral genome. The gene sequences of the viruses obtained from the two different fish species shared 94.4% nucleotide sequence identity (98.1% amino acid sequence identity), suggesting that they belong to the same virus species. Phylogenetic analysis based on the DNA polymerase (and the concatenated sequences of the amplified genes, as well) implied that the detected virus belongs to the genus Cyprinivirus within the family Alloherpesviridae. The sequences of the novel alloherpesvirus diverge from those of the five cyprinivirus species described previously, so it putatively represents the sixth virus species in the genus.

Infection of the Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) with a dermocystid parasite in the Tisza River Basin, Hungary.

The Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) is an endemic protected species of Cephalaspidomorphi in the Carpathian Basin. No parasites have become known from these jawless vertebrates to date. Here, the authors describe an infection from a single specimen manifesting in protuberant skin cysts 7-10 mm in diameter, scattered on the body surface. Similar dermal infection was observed in 25 of the 274 lampreys recorded in the population survey. Skin cysts filled with round spore-like structures of a dermocystid parasite were found. These particles measured 8-14 µm in diameter and had an about 0.5 µm thick wall, and containing mainly a granular mass and a relatively scarce plasma. No hyphae were recorded. Despite conspicuous morphological changes in the skin, no inflammatory reactions were found. The molecular analysis of 18S rDNA showed similarity to dermocystid species of several fish species but differed from them approximately by 2%. This is the first record of a dermocystid parasite infecting a jawless vertebrate.

New record of metacercariae of the North American Posthodiplostomum centrarchi (Digenea, Diplostomidae) in pumpkinseed (Lepomis gibbosus) in Hungary.

Two species of the genus Posthodiplostomum (Digenea: Diplostomatidae) (Posthodiplostomum brevicaudatum Nordmann, 1832 and Posthodiplostomum cuticola Nordmann, 1832) are known as parasites of Hungarian native fishes. Metacercariae of P. cuticola are widespread in Europe and cause black spot disease. Several species of Posthodiplostomum were described also from North America but none of them has been isolated in Hungary up to now. Posthodiplostomum centrarchi Hoffman, 1958 has been detected recently in pumpkinseeds (Lepomis gibbosus L., 1758) in several European countries. Posthodiplostomum centrarchi was isolated for the first time in Hungary from pumpkinseeds caught in the Maconka water reservoir in 2015. Thereafter, several natural waters (e.g. the River Danube, Lake Balaton and the Sió channel) were sampled in order to determine its presence and distribution. Only the native species P. cuticola was detected in Lake Balaton on cyprinids but a relatively high infection rate of P. centrarchi was observed in the Sió channel close to the lake. Pathological changes were absent, and metacercariae were mostly attached to the surface of the liver, kidney and heart. The phylogenetic analysis of the ITS and COI sequences of P. centrarchi and P. cuticola clustered into two distinct branches, which was in agreement with the morphological results.

Cross section of gill filaments in histological preparations helps better identification of the location of myxosporean plasmodia in gill tissues.

Location and tissue preference of filamental-type myxosporean plasmodia in histological slides of the gills can be properly identified only in cross sections of the gill filaments. The authors selected three myxosporeans (Myxobolus rutili, M. dispar and Henneguya psorospermica, parasites of the roach, the common carp and the pike, respectively) for studying the problem. The plasmodia of these species studied in longitudinal sections were earlier regarded as developing inside the filamental arteries. Cross sections of the filaments showed that all the three species developed plasmodia in the dense connective tissue constituting the adventitia of gill arteries and covering the cartilaginous gill rays. Myxobolus rutili started its development close to the afferent branchial artery but attached to the cartilaginous gill ray. More developed plasmodia of this species surrounded the rays. Plasmodia of M. dispar were formed on the inner side of the afferent branchial artery, while those of H. psorospermica were located at the external side of the efferent branchial artery.

[Early experience with minimal invasive treatment of thirty-one thoracolumbar injury cases]. / Korai tapasztalataink thoracolumbalis gerincsérülések minimálinvazív mutéti kezelésével.

The authors report on their experience with minimal invasive treatment of thirtyone thoracolumbar injury cases. The medical charts, radiological documentation (preop CT and MRI scans, postop CT scans at 3 and 6 months) were analysed retrospectively. All pedicle screws were inserted appropriately, there was no neurological deterioration or surgical site infection postoperatively. There were three cases of radiological loss of correction, however, these were clinically irrelevant. There were no screw loosening or implant failure observed. According to our experience, which coincedes with literature data, this minimally invasive percutaneous pedicle screw technique can be recommended for the treatment of thoracolumbar injury.

Histological investigation on Ancyrocephalus paradoxus (Dactylogyridea: Ancyrocephalidae) infection causing mortalities in an intensively cultured pikeperch [Sander lucioperca (L.)] stock.

In a cultured pikeperch (Sander lucioperca) stock the monopisthocotylean monogenean gill parasite Ancyrocephalus paradoxus caused heavy infection and mortalities. The gills of the affected fish specimens were infected by 50 to 800 monogenean parasites. Severe pathological changes were found in areas where the worms attached to the gills. At the attachment sites the haptoral discs of the worms formed a deep depression in the epithelium of the filaments, and the anchors pierced into and fixed themselves to the connective tissue of the cartilaginous gill rays. At these attachment sites red blood cells released from injured capillaries were found among the damaged epithelial cells. Around the hooks, anchors and body sections coming into contact with the gill filaments a proliferative tissue developed in which only a remnant of the damaged lamellae was found. Due to the damage caused by the worms the tips of the heavily infected gill filaments fused, formed clubs and were composed of epitheloid-type regeneration tissue lacking respiratory lamellae. In the basal parts of the filaments, where most of the worms attached to the gill, only denuded filaments deprived of lamellae were observed among the cross-sectioned worms in histological sections.

Molecular genetic investigations on Balantidium ctenopharyngodoni Chen, 1955, a parasite of the grass carp (Ctenopharyngodon idella).

Balantidium ctenopharyngodoni is a common ciliate in Hungary, infecting the hindgut of grass carp (Ctenopharyngodon idella), a cyprinid fish of Chinese origin. Although data have already been presented on its occasional pathogenic effect on the endothelium of the host, generally it is a harmless inhabitant of the gut. Phylogenetic analysis of the 18S rDNA and ITS fragments of this protozoan proved that it is in the closest phylogenetic relationship with endocommensalist and symbiont ciliates of mammals feeding on large volumes of green forage, in a similar way as Balantidium spp. known from algae-eating marine fishes.

[Our innovations for the reduction of postoperative pancreatic fistula output]. / A posztoperatív pancreasfistula csökkentésére bevezetett innovációink.

Bevezetés: A posztoperatív pancreasfistula mind proximalis, mind distalis pancreatectomia után a legjelentosebb sebészi szövodménynek számít. A szakirodalomban nincs egyértelmuen ajánlott, megbízható módszer ezen probléma kiküszöbölésére, emiatt történnek újítások szerte a világon. Jelen közleményünkben a technikai innovációinkról számolunk be. Anyag és módszerek: 2013. január 1-jétol 2023. november 30-ig terjedo idoszakban 205 Whipple-mutétet végeztünk nyitottan, mely során a pancreatojejunalis anastomosist az általunk módosított dohányzacskó-öltéses módszerrel készítettük el. 2019. január 1. és 2023. november 30. között pedig 30 betegnél történt nyitott distalis pancreatectomia, amikor a pancreascsonkot az általunk kifejlesztett technikával, szabad rectus fascia-peritoneum grafttal fedtük, majd azt cirkuláris öltéssel rögzítettük. Közleményünkben ezen két módszerrel elért eredményeket ismertetjük. Eredmények: a demográfiai adatok megfeleltek a betegségnél szokásosnak. A posztoperatív ápolási ido és a transzfúzió igény terén észlelt különbségek tükrözték a kétféle beavatkozás eltéro invazivitását. A releváns pancreasfistula kialakulási rátája kedvezo képet mutatott, Whipple-mutét után 7,3% volt, míg distalis pancreatectomát követoen nem fejlodött ki. A reoperációs és a halálozási arányok megfeleltek az elvártaknak és korreláltak a mutétek kiterjedtségével. Következtetés: pancreas resectiók utáni komplikációk csökkentésére tett törekvéseink során a módosított dohányzacskó-öltéses pancreatojejunostomia és a pancreascsonk fedésére kidolgozott módszerünk egyaránt kedvezo eredményekkel járt.

Interpretation of LDH Values after Kidney Transplantation.

Kidney transplantation is the gold-standard therapy for end-stage renal disease. However, in the early postoperative period following allograft kidney transplantation, insufficient graft function presents a diagnostic challenge to clinicians. Ischemic damage to the graft and/or an early autoimmune rejection may cause a decrease in function. Ischemic damage is a benign and transient condition, while acute immune rejection requires immediate therapy. A kidney graft ultrasound may produce a false negative result, and graft biopsy is invasive and slow to return results. Serum lactate dehydrogenase (LDH) is under examination as a possible tool for differential diagnosis between ischemic damage and immune rejection. Herein, we analyze the continuous lab results of four patients in the early post-transplantation period, showing patterns correlating with different clinical outcomes and prognoses. In our experience, a persistent elevated LDH accompanies ischemic damage. Immune rejection was, however, associated with a decrease in LDH. Hemodialysis was not a confounding factor, while packed red blood cell transfusion caused severe diagnostic problems.

Controlled lumbar cerebrospinal fluid drainage effectively decreases the need for second and third tier interventions for intracranial hypertension in severe traumatic brain injury patients.

INTRODUCTION: Early treatment of elevated intracranial pressure (ICP) is a cornerstone of the therapy in severe traumatic brain injury (TBI) patients. Treatment of refractory high ICP however, remain challenging as only limited and risky third-tier therapeutic interventions are available. Controlled lumbar cerebrospinal fluid (CSF) drainage has been known as an efficient method of ICP reduction after TBI for decades, but it is not recommended in international guidelines because of low evidence background and safety issues. Our centre has a long-standing experience using this intervention for more than 15 years. Here we present our data about the safety and efficacy of controlled lumbar drainage to avoid further second- and third tier ICP lowering therapies and beneficially influence functional outcome. METHODS: Observational (retrospective and prospective) analysis was performed using demographic, clinical and outcome data of severe TBI patients admitted to our centre. Analysis was retrospective between 2008 and 2013 and prospective from 2014 to 2019. Only severe TBI patients (GCS<9) with ICP monitoring were enrolled. Lumbar drainage (LD) was used as a second-tier therapy to control intracranial hypertension in salvageable patients with normal haemostasis and discernible basal cisterns on pre-interventional CT scan. RESULTS: Data of 45 patients were analysed. Patients were young, comatose and with severe injuries (median age: 29, GMS: 4, ISS: 25). Lumbar drain was inserted mainly on the first week and maintained for further 5 days. Episodes of intracranial hypertension (ICP>20 Hgmm) within one day (10 vs 2) were reduced. The need of additional second- and third-line therapies (deep sedation, hyperventilation, barbiturate administration, decompressive craniectomy) also significantly decreased (60 vs 25 interventions, p<0.001). The in-hospital mortality and 6-month functional outcome were more favourable than the whole TBI population and as predicted by prognostic calculations (mortality: 16% vs. 48 %; GOSE 1-4: 49% vs. 65% vs CRASH: 87% vs. IMPACT: 51 %) in this period. CONCLUSIONS: Our results support the view that controlled lumbar drainage is a highly efficient method to manage intracranial hypertension and significantly decreases the need of further harmful ICP lowering therapies without altering functional outcome of severe TBI patients.

Effect of Bile on Hemodynamics and Blood Micro-Rheological Parameters in Experimental Models of Bilhemia.

As a rare complication of liver injury and certain interventions, bile can enter the bloodstream depending on the pressure gradient, resulting in bilhemia. Its micro-rheological and hemodynamic effects are still unclear. We aimed to study these parameters in experimental bilhemia models. Under general anesthesia, via laparotomy, bile was obtained by gallbladder puncture from pigs and by choledochal duct cannulation from rats. In vitro, 1 µL and 5 µL of bile were mixed with 500 µL of anticoagulated autologous blood. The systemic effect was also assessed (i.v. bile, 200 µL/bwkg). Hemodynamic and hematological parameters were monitored, and red blood cell (RBC) deformability and aggregation were determined. RBC deformability significantly decreased with the increasing bile concentration in vitro (1 µL: p = 0.033; 5 µL: p < 0.001) in both species. The RBC aggregation index values were concomitantly worsened (1 µL: p < 0.001; 5 µL: p < 0.001). The mean arterial pressure and heart rate decreased by 15.2 ± 6.9% and 4.6 ± 2.1% in rats (in 10.6 ± 2.6 s) and by 32.1 ± 14% and 25.2 ± 11.63% in pigs (in 48.3 ± 18.9 s). Restoration of the values was observed in 45 ± 9.5 s (rats) and 130 ± 20 s (pigs). Bilhemia directly affected the hemodynamic parameters and caused micro-rheological deterioration. The magnitude and dynamics of the changes were different for the two species.

Removal of dead fish eggs by Asellus aquaticus as a potential biological control in aquaculture.

The objective of the present experimental study was to gain a better understanding of the foraging activity of Asellus aquaticus during fish egg incubation. A. aquaticus were introduced into experimental setups of dead eggs, viable eggs and hatched larvae of zebrafish (Danio rerio), a commonly used model organism. The amount of A. aquaticus and the duration of their exposure to the eggs significantly affected the proportion of consumed dead eggs in each experimental cycle. A. aquaticus belongs to the group of aquatic detritivores, and no predatory behavior was observed during the experiments. These crustaceans could distinguish between the dead eggs and those containing living embryos. Furthermore, zebrafish larvae remained unharmed by A. aquaticus, even in the absence of alternative food source. These findings underscore the potential sanitary role of these crustaceans in natural waters and offer new perspectives on their possible use as biological control organisms in aquaculture hatcheries. Additionally, our results suggest a potential application of A. aquaticus in combating pathogens by reducing the growth substrates for bacteria and fungi.