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Colorectal cancer patients treated with irinotecan (IRI) often manifest acute cholinergic syndrome, which is the reason behind the growing use of legally prescribed cannabis preparations as well as illicit ones that can contain very high Δ9- tetrahydrocannabinol (THC) levels. IRI-induced cholinergic syndrome originates from the inhibition of acetylcholinesterase (AChE), while butyrylcholinesterase (BChE) plays an important role in the hydrolytic metabolism of this prodrug. On the other hand, existing reports of THC effects on cholinesterases are inconclusive. In this study, we aimed to investigate how the concomitant use of a high THC dose with IRI affects cholinesterase activities in blood of mice with experimental colon cancer induced by injection of CT26 cell line. Male BALB/c mice were assigned randomly to five study groups: (i) negative control, (ii) cancer control, (iii) irinotecan (i.p. 60 mg/kg ; 1st and 5th day of the experiment), (iv) THC (p.o. 7 mg/kg ; repeatedly for 7 days), (v) IRI + THC. ChE activity in blood was assayed by the spectrophotometric Ellman method, using acetylthiocholine as a substrate and inhibitors to distinguish between AChE and BChE activities. The results showed a time-dependent increase in total ChE and BChE activities in all experimental groups, including cancer control. In contrast, AChE activities decreased, with some minor fluctuations in the THC and IRI+THC group. Although there was a lack of statistically significant differences, the time course of changes for AChE and BChE activities suggested that these esterases may not only be indicators of metabolic status but also functionally important in neoplastic cell transformation.

Irinotecan (IRI) represents one of the most important antineoplastic drugs used in the chemotherapy of metastatic colorectal cancer. Its use is accompanied by various adverse effects such as diarrhoea, myelosuppression and hepatotoxicity. To diminish side-effects, some cancer patients use Cannabis sativa preparations, rich on delta-9-tetrahydrocannabinol (THC). The potentially harmful interactions resulting from co-administration of chemotherapy with biologically based complementary therapies have not yet been explained well, which motivated us to conduct a study on a Wistar rat model using a THC dose equal to the one found in commonly used illicit preparations. Male rats were concomitantly exposed to IRI (at 100 mg/kg b.w., administered once i.p.) and THC (administered repeatedly for 3 and 7 days per os at 7 mg/kg b.w.). Single IRI, THC and control groups were studied in parallel. We estimated changes in brain weight caused by the treatments and evaluated the level of primary DNA damage in the brain cells of exposed and control rats using alkaline comet assay. Combined treatment slightly diminished brain weights compared to controls after both exposure times. The 3-day treatment resulted in significantly increased levels of DNA damage in brain cells of rats given single IRI, which worsened after concomitant exposure to THC. Single THC caused a minor increase of DNA damage compared to control rats. After the 7-day treatment, levels of DNA damage in rats given single IRI and IRI+THC slightly lowered, but were still significantly higher than in control rats. Prolonged exposure to THC resulted in a further increase of DNA damage. The obtained results speak against the concomitant use of THC with IRI, due to the possible enhancement of brain damage, but these findings have to be proven in forthcoming studies using much wider dose ranges of both compounds.

Normal saline or 0.9% NaCl solution is the most commonly used intravenous fluid worldwide and its composition are 154 mmol Na+ and 154 mmol Cl- per litter with osmolarity of 308 mOsmol/L. That composition is not “normal” because plasma contains sodium in range between 137-146 mmol/L, and chloride in range between 98-106 mmol/L, with plasma osmolality of 280- 295 mOsmol/kg. Recently published data suggested detrimental effects of chloride rich fluids on renal blood flow and glomerular filtration rate, urine output and acute kidney injury. One alternative to saline solution is a buffered, balanced, crystalloid solution with an electrolyte composition similar to plasma and osmolarity between 286-295 mOsmol/L. Someone could indicate that such balanced solutions are not suitable for neurosurgical patients because of a possible impact on the brain oedema development.

During the past decades, several scoring systems have been developed to assess the functional status of patients with spine related problems. Degenerative spine diseases are the most frequent cause of disability and work incapacity, which represents large social and economic problem in developing countries. There are surgical and non-surgical treatments and concurrently growing demand for outcome measurement and quality assurance. According to this we used economic and functional rating score (Prolo scale) and VAS pain scale before and after spine operation, as well as EQ-5D-5L (Euroqol Haelth Questionnaire) after more than 6 months of surgery. This observational study is the first step in creation of national strategy for prevention and treatment such conditions.

We report a case of chronic intra-abdominal hypertension in patient with giant liposarcoma and finally its presentation as inferior vena cava syndrome.

Meningitis in neurosurgical patients is serious postoperative complication that needs early recognition and treatment. Diagnosis of meningitis in the neurosurgical patients might be difficult because of overlapping clinical presentation caused by surgery or trauma. Standard CSF studies are unreliable because of blood caused by subarachnoid haemorrhage (SAH) or surgical trauma. The aim of this retrospective study is to evaluate and compare the diagnostic usefulness of cerebrospinal fluid (CSF) obtained from external ventricular derivation (EVD) and lumbar puncture (LP) for detection of postoperative bacterial meningitis.

Organophosphorus (OP) compound poisoning is an important worldwide clinical and public health problem. Some nerve warfare agents have relatively recently been used in terrorist attacks, while pesticide poisonings represent the leading cause of poisonings reported globally. OPs act primarily as neurotoxins due to the irreversible inhibition of acetylcholinesterase (AChE), but there is increasing evidence for secondary mechanisms of their toxicity. Oximes act as an essential part of antidotal therapy by ensuring the recovery of inhibited AChE. Our recent in vitro and in vivo studies have drawn attention to oxime 1- (4-hydroxyiminomethylpyridinium)-4-(4- carbamoylpyridinium)-butane dibromide (K048) as a very potent reactivator of the nerve agent tabun- and pesticide-inhibited AChE. This study evaluated OP toxicity at molecular level and its potential alleviation by the use of an oxime. For this purpose, as it is one of the most potent AChE-inhibiting compounds available, we selected paraoxon as a model OP and estimated its ability to induce primary DNA damage in two human cell lines known for non- neuromuscular AChE expression. By applying the K048 oxime, we tested its ability to extenuate the effects of paraoxon. Human lung adenocarcinoma epithelial cell line A549 and human keratinocyte cell line HaCaT were cultivated in RPMI growth medium supplemented with glutamine, heat-inactivated fetal bovine serum, and antibiotics. Prior to treatment, cells were seeded in 6-well plates (3x105 cells/mL). The tested concentrations of paraoxon (1 µM), and K048 oxime (0.02 mM) were determined in a preliminary experiment on human erythrocytes. For the genotoxicity testing, we applied the same study design in both cell lines: (1) control group (untreated cells), (2) cells treated with paraoxon (for 10 min), (3) cells treated with K048 (for 30 min), (4) cells treated 10 min with paraoxon followed by a 30- min treatment with K048 oxime. After the treatments, aliquots of cells were used for the preparation of agarose microgels according to standard protocol for the alkaline comet assay. Slides were analyzed under fluorescent microscope using the Comet Assay IV analysis system (Perceptive Instruments Ltd., UK). The level of DNA damage was evaluated based on comet tail length, tail intensity, and tail moment. AChE activity measurements were performed by using the standardized spectrophotometric Ellman method. At the concentration tested, the K048 oxime did not induce a significant increase of primary DNA damage as compared to the negative control. In contrast, treatment with paraoxon, in spite of the relatively short exposure time, resulted in measurable genotoxicity both on A549 and HaCaT cells. At the same concentration, paraoxon caused a 93% erythrocyte AChE inhibition in our preliminary experiment. The most important observation was the lowering of primary DNA damage in cells treated with both compounds. In both cell lines, when applied after paraoxon, K048 caused a significant decrease of all comet assay parameters as compared to single paraoxon treatment. Furthermore, K048 was shown to reactivate about 87% of paraoxon-inhibited erythrocyte AChE. Apart from its high reactivation potency at the concentration tested, the K048 oxime also possesses antioxidative efficacy, which is an important property for a compound intended for use as an antidote.

Aim: Oximes play an important role in the elimination of toxic effects caused by chemical warfare agents, especially in the cases resistant to the atropine treatment alone. K048 oxime as one of the promising antidote in humans recently showed an acceptable genotoxic profile. Present study is focused on potential hepatoprotective effects of K048 effects in vivo. Methods: We selected a model of tabun- poisoned rats, which were given K048 oxime to relieve symptoms of tabun-poisoning. The levels of primary DNA damage in liver cells were studied using the alkaline comet assay 1, 6 and 24 h after the treatments. Sixty male adult Wistar rats were divided in five groups according to the treatment: (1) tabun-poisoned (75% LD50, s.c.), (2) K048-treated (25% LD50 i.p.), (3) atropine-treated (10 mg kg-1, i.p.) after tabun poisoning (75% LD50, s.c.), (4) concomitant treatment with K048 (25% LD50, i.p.) and atropine (10 mg kg-1, i.p.) after tabun poisoning (75% LD50, s.c.), and (5) negative controls given saline only (2 mL kg-1, i.p.). Results: Tabun itself caused the highest level of primary DNA damage in hepatocytes 24 h after its administration. K048 oxime had an acceptable genotoxicity profile at each time point. Its hepatoprotective effects were obvious at each time point studied, and in all cases the levels of primary DNA damage measured in oxime-treated groups were lower or similar as compared to negative control. Minor differences in the genotoxicity observed between the time-points studied could be explained by the metabolism of the compound. Concomitant administration of K048 with atropine changed its genotoxicity profile in the hepatocytes, especially 6 h after treatment. Conclusions: K048 effectively counteracted the poisoning of rats by tabun, both alone and in combination with atropine. Hepatoprotective effects speak in favour of K048 as a promising molecule for therapeutic treatment against fatal poisoning with chemical warfare agents.

Organophosphorus (OP) compounds cause fatal intoxication based on inhibition of acetylcholinesterase (AChE), an essential enzyme in neurotransmission. There is an obvious need for appropriate therapeutic treatment against them due to their application in agriculture or their misuse in terrorist or war attack. Pyridinium oximes are causal antidotes employed for the protection/reactivation of AChE inhibited by OPs. Although their pharmacological profile is well known, possible cyto- and genotoxic effects have not been studied in detail so far. In present in vivo study we wanted to elucidate the toxicity profile of the standard oxime HI-6 and its modification - oxime K048 by simultaneous use of biochemical and genotoxicity assays. The catalytic activities of total cholinesterases (ChE) in plasma and of AChE in brain were determined 30 min, 60 min and 24 h after the rats were administered with single therapeutic dose of oximes (25% of their LD50). In addition, blood and brain tissue were sampled 24 h after treatment and used for the alkaline comet assay to study the levels of primary DNA damage in white blood cells (WBC) and brain cells. Experimental animals did not exhibit any clinical signs of poisoning following application of both oximes. Up to 25.3% decrease in plasma and up to 18.5% decrease in brain enzyme activities was observed in all experimental animals after application of HI-6. A decrease of enzyme activities after application of K048 was noticed only for time-point at 60 min in both plasma (11.9%) and brain (20%). These results indicate higher affinity of intact blood and tissue AChE for HI-6 but there were no significant differences compared to corresponding controls. Treatment with HI-6 and K048 did not induce a significant increase of DNA damage compared to the corresponding controls. Mean tail DNA% in the WBC of HI-6 treated rats was 0.27±0.03% (range: 0-3.90%), and in the brain cells 0.99±0.06% (range: 0-4.87%). Treatment with K048 induced slightly higher levels of primary DNA damage, compared to HI-6, both in the WBC (mean tail DNA% 0.56±0.07, range: 0-8.11%) and in the brain cells (mean tail DNA % 1.14±0.14, range 0-21.27%). Although both oximes had acceptable genotoxicity profiles, HI-6 induced lower level of DNA damage in the brain cells than K048 ; there were 6% of highly damaged comets with respect to the tail DNA% in HI-6 treated rats vs. 10% of highly damaged comets with respect to the tail DNA% in K048 treated rats. Taken together, results obtained by the enzyme assay and the alkaline comet assay, speak in favour of HI-6 as a promising molecule for therapeutic treatment against intoxication with OP compounds.

Congenital hyperinsulinism (CHI) is a major cause of persistent hypoglycemia in the neonatal and early infancy periods. Althought the disease is relatively rare with incidence of about 1:25 000-50 000 live births, the importance of the disease should not be underestimated. Namely, prompt recognition and management of patients with CHI is essential, if permanent neurological impairment is to be avoided. CHI is caused by mutations in one of the 7 genes involved in the regulation of insulin secretion in pancreatic beta-cells. It is important to introduce specific medical therapy as soon as diagnosis is established. Severe, neonatal forms of CHI are often resistant to medications, thus they require surgical procedure. The preoperative genetic testing and scintigraphy are indicated to distinguish histological subtypes of the disease (focal vs. diffuse CHI). Patients with focal disease are usually cured after pancreatic resection, while diffuse disease has much worse prognosis. This manuscript offers novel insights into CHI and emphasizes the role of early diagnosis as crucial for succesful treatment that was recently enriched with novel options.

Kongenitalni hiperinzulinizam (KHI) najčešći je uzrok tvrdokornih hipoglikemija u novorođenačkom i ranome dojenačkom razdoblju. Iako je bolest razmjerno rijetka s pojavnosti od oko 1 : 25 000–50 000 živorođene djece, bolest se ne smije podcijeniti zbog trajnih neuroloških oštećenja do kojih dolazi ako se bolest brzo ne otkrije i ne započne promptno liječiti. Uzrok su mutacije nekog od 7 gena, ključnih u regulaciji inzulinske sekrecije u b-stanicama gušterače. Odmah nakon postavljanja dijagnoze nužno je u terapiju uvesti antihipoglikemike koji djeluju specifično na -stanice gušterače. Kod teških neonatalnih oblika nerijetko postoji rezistencija na antihipoglikemike. Tada preostaje kirurško liječenje prije kojeg je potrebno odrediti histološki oblik bolesti, što se danas uspješno postiže kombinacijom genskog testiranja i scintigrafske pretrage. Kirurškim se zahvatom najčešće postiže izlječenje kod fokalnog KHI, dok difuzni oblik ima lošiju prognozu. Ovaj je tekst pregled novijih spoznaja o KHI, kojim želimo naglasiti važnost što ranijeg postavljanja dijagnoze kao preduvjeta uspješnoga ciljanog liječenja bolesti, koje je posljednjih godina obogaćeno novim mogućnostima.

Pediatric liver transplantation in Croatia

Outcome of children with esophageal atresia – 15 years experience

U ekstrahepatalnoj portalnoj hipertenziji shunt je dugoročno uspješna metoda liječenja

12 bolesnika operiranih zbog tumora jetre, prikaz vrste i ishoda

Outcome of postoperative treatment in newborns with congenital anomalies of digestive system and abdominal wall - 16 years of experience