POSSIBILITY OF NEW BIOTECHNOLOGY ACHIEVEMENTS IN IMPROVEMENT OF MILK QUALITY

Novija biotehnološka dostignuća ušla su u sve segmente animalne proizvodnje, pa tako i u proces proizvodnje mlijeka. Interes uzgajivača je unaprijediti genetsku osnovu i tehnologiju kako bi što učinkovitije proizveli kvalitetno mlijeko i ostvarili primjeren profit. Primjenom biotehnologija otvaraju se mogućnosti pomoću kojih je moguće napraviti korak naprijed u selekciji, unaprijediti genetsku osnovu, te uz kvalitetna grla proizvesti mlijeko koje će po svome sadržaju i po kvaliteti odgovarati točno određenim zahtjevima u proizvodnji određenog tipa i vrste mliječnih proizvoda. Od biotehnoloških metoda koje se češće koriste su embrio transfer i seksirano sjeme kvalitetnih bikova. Metode se rabe u funkciji unapređenja uzgojnog programa kao i praktične selekcije (unapređenja) stada. U radu je opisana tehnologija embriotransfera, seksiranog sjemena te polimorfizama proteina mlijeka i njihov utjecaj na količinu mlijeka, mliječne masti i proteina kao i na sposobnost mlijeka za preradu.Recent biotechnology advances have entered into all segments of animal production, including the process of milk production. The breeders are interested into improving the genetic base and technology in order to produce quality milk more effectively and make a suitable profit. Applying biotechnology opens the opportunities with which it is possible to move forward in the selection process, to improve the genetic basis, and produce milk which will, due to its content and quality, match the specific requirements in the production of a certain types and sorts of dairy products. Biotechnological methods that are often used are embryo transfer and semen sexation of high-quality bulls. Methods are used in order to improve the breeding program, as well as the practical selection (promotion) of the herds. This paper describes the embryo transfer technology, semen sexation, as well as milk protein polymorphisms and their effect on milk yield, milk fat and protein, including the ability of milk processing

Use of biochemical parameters for non-invasive screening of oesophageal varices in comparison to elastography-based approach in patients with compensated advanced chronic liver disease

Oesophageal varices are routinely diagnosed by esophagogastroduodenoscopy (EGD), and their bleeding has high mortality. We aimed to evaluate diagnostic performance of biochemical tests in comparison to elastography-based approaches, as non-invasive alternatives to EGD, for ruling-out high risk oesophageal varices (HRV). Retrospective analysis of patients (N = 861) who underwent liver stiffness measurement (LSM) by transient elastography (TE) in a single centre over 5-year period, with available results of EGD (within 3 months from LSM). Only patients with suspicion of compensated advanced chronic liver disease (cACLD) defined by LSM ≥ 10 kPa were included comprising the final cohort of 73 subjects. Original and expanded Baveno VI criteria (B6C), controlled attenuation parameter (CAP), platelet count (PLT), aspartate aminotransferase to PLT ratio index (APRI), Fibrosis-4 index (FIB4), model for end stage liver disease (MELD) score were evaluated against the results of EGD that served as the reference method. Analysed patients had median age 62 years, 59/73 (0.81) were males, 54/73 (0.74) had alcoholic/non-alcoholic fatty liver disease, and 21/73 (0.29) had HRV. In multivariate logistic regression analysis only LSM and PLT were independently associated with HRV. The best performing tests for ruling-out HRV (% of spared EGD; % of missed HRV) were respectively: LSM 214x109/L (21.9%; 0%); FIB4 ≤ 1.8 (21.4%; 0%), APRI ≤ 0.34 (12.3%; 0%). CAP, MELD = 6 alone or combined with PLT > 150(x109/L) did not show acceptable performance. The best performing biochemical tests for ruling-out HRV in our cohort of patients were PLT and FIB-4, but they were still outperformed by elastography-based approaches

LiverScreen project: study protocol for screening for liver fibrosis in the general population in European countries

Background: The development of liver cirrhosis is usually an asymptomatic process until late stages when complications occur. The potential reversibility of the disease is dependent on early diagnosis of liver fibrosis and timely targeted treatment. Recently, the use of non-invasive tools has been suggested for screening of liver fibrosis, especially in subjects with risk factors for chronic liver disease. Nevertheless, large population-based studies with cost-effectiveness analyses are still lacking to support the widespread use of such tools. The aim of this study is to investigate whether non-invasive liver stiffness measurement in the general population is useful to identify subjects with asymptomatic, advanced chronic liver disease. Methods: This study aims to include 30,000 subjects from eight European countries. Subjects from the general population aged ≥ 40 years without known liver disease will be invited to participate in the study either through phone calls/letters or through their primary care center. In the first study visit, subjects will undergo bloodwork as well as hepatic fat quantification and liver stiffness measurement (LSM) by vibration-controlled transient elastography. If LSM is ≥ 8 kPa and/or if ALT levels are ≥1.5 x upper limit of normal, subjects will be referred to hospital for further evaluation and consideration of liver biopsy. The primary outcome is the percentage of subjects with LSM ≥ 8kPa. In addition, a health economic evaluation will be performed to assess the cost-effectiveness and budget impact of such an intervention. The project is funded by the European Commission H2020 program. Discussion: This study comes at an especially important time, as the burden of chronic liver diseases is expected to increase in the coming years. There is consequently an urgent need to change our current approach, from diagnosing the disease late when the impact of interventions may be limited to diagnosing the disease earlier, when the patient is asymptomatic and free of complications, and the disease potentially reversible. Ultimately, the LiverScreen study will serve as a basis from which diagnostic pathways can be developed and adapted to the specific socio-economic and healthcare conditions in each country

Characteristics of patients with primary biliary cholangitis in a tertiary hospital facility

Uvod: Primarni bilijarni kolangitis (PBC) je progresivna, autoimuna bolest jetre, kroničnog tijeka i nedovoljno poznate etiologije. Bolest se aktivira kada osobe s genetskom predispozicijom budu izložene štetnim čimbenicima iz okoliša. U zadnje vrijeme, opisuje se porast prevalencije i incidencije primarnog bilijarnog kolangitisa u cijelom svijetu. Epidemiološki gradijent pokazuje pad prevalencije u smjeru zapad-istok i sjever-jug. Trenutno nema objavljenih podataka o incidenciji, prevalenciji ili demografskim značajkama oboljelih od PBC-a u Hrvatskoj. Glavni cilj ovog istraživanja bio je prikazati epidemiološke, kliničke i demografske karakteristike PBC bolesnika liječenih u Kliničkoj bolnici Dubrava. ----- Metode: Provedeno je retrospektivno istraživanje bolničke baze podataka za period od 2007. do 2017. godine. Identificirano je 46 PBC pacijenata s dostupnom medicinskom dokumentacijom. Analizirani su demografski, klinički i epidemiološki podatci, odgovor na terapiju nakon 12 mjeseci korištenja, te klinički ishod bolesti. Lošim kliničkim ishodom smatrani su: smrt, razvoj raka jetre, potreba za transplantacijom jetre ili jetrena dekompenzacija. Za procjenu incidencije i prevalencije uzeti su u obzir samo bolesnici s prebivalištem na geografskom području odgovornosti KB Dubrava. ----- Rezultati: Uključeni bolesnici bili su pretežito ženskog spola (87%) s medijanom starosti od 57,5 godina, a 68,8% ih je bilo u ranom kliničkom stadiju bolesti. Analizom bolesnika s prebivalištem na području odgovornosti KBD (N=38) izračunata je prevalencija od 11,47 bolesnika na 100,000 stanovnika i godišnja incidencija od 0,54 na 100,000 stanovnika. Svi bolesnici liječeni su ursodeoksikolnom kiselinom (UDCA). Među 32 bolesnika za koje su bili dostupni podatci nakon 12 mjeseci terapije, povoljan biokemijski odgovor zabilježen je u njih 25 (78%). Čimbenici povezani s uspjehom liječenja bili su redovito uzimanje terapije (P = 0,007) i niska inicijalna koncentracija IgM-a (P = 0,009). Loš klinički ishod zabilježen je u 5 bolesnika. U multivarijatnoj analizi osteoporoza je bila jedini čimbenik neovisno povezan s lošim ishodom bolesti (HR = 9,27; P = 0,031). ----- Zaključak: PBC je rijetka bolest, koja u opisanoj kohorti pogađa pretežito žene, srednje dobi, a većina ih ima blagi klinički stadij i dobru prognozu. Iako zemljopisno ograničeno, ovo istraživanje pruža prve epidemiološke podatke o PBC-u u Hrvatskoj i time nadopunjuje PBC kartu Europe.Introduction: Primary biliary cholangitis (PBC) is a chronic and progressive cholestatic liver disease of unknown etiology. Nowadays, PBC is considered an autoimmune disease activated by environmental factors in individuals with a genetic predisposition. Prevalence and incidence rates of PBC appear to increase worldwide. The epidemiological gradient shows a decline in prevalence from West to East and from North to South. There are no published data on incidence, prevalence or demographic characteristics of patients with primary biliary cholangitis (PBC) in Croatia. The aim of this study was to show the epidemiological and demographic characteristics of PBC patients treated at Dubrava Clinical Hospital. ----- Methods: A retrospective search of the hospital´s database was conducted. Data from 2007 to 2017 were reviewed during the study. 46 PBC patients with retrievable medical records were identified. Epidemiologic data were restricted to patients from the hospital´s catchment area. Statistical analysis involving the entire cohort included descriptive data as well as response to therapy after 12 months and a survival analysis. Risk factors for an adverse outcome were identified. An adverse outcome was defined as a composite event (ascites, variceal bleeding, encephalopathy), or HCC, liver transplantation and death. ----- Results: Most patients were female (87%) with a median age of disease onset of 57.5 years. The cohort included 38 patients (83%) from the hospital´s catchment area resulting in the prevalence of 11.47/100.000 and incidence of 0.54/100.000 inhabitants per year. From 32 patients analyzed concerning the response to therapy, 78% responded well. Adherence to therapy (P=0.007) and low IgM concentration at diagnosis (P=0.009) were associated with therapy success. An adverse outcome was reported in five patients. Among factors related to shorter event-free survival, osteoporosis remained relevant in multivariate analysis after adjusting for age (HR=9.27; P=0.031). ----- Conclusion: Despite some drawbacks, this study gives pioneer insights into the epidemiological data of PBC in the southeast corner of Europe, therefore complementing the PBC map of Europe

Characteristics of patients with primary biliary cholangitis in a tertiary hospital facility

Uvod: Primarni bilijarni kolangitis (PBC) je progresivna, autoimuna bolest jetre, kroničnog tijeka i nedovoljno poznate etiologije. Bolest se aktivira kada osobe s genetskom predispozicijom budu izložene štetnim čimbenicima iz okoliša. U zadnje vrijeme, opisuje se porast prevalencije i incidencije primarnog bilijarnog kolangitisa u cijelom svijetu. Epidemiološki gradijent pokazuje pad prevalencije u smjeru zapad-istok i sjever-jug. Trenutno nema objavljenih podataka o incidenciji, prevalenciji ili demografskim značajkama oboljelih od PBC-a u Hrvatskoj. Glavni cilj ovog istraživanja bio je prikazati epidemiološke, kliničke i demografske karakteristike PBC bolesnika liječenih u Kliničkoj bolnici Dubrava. ----- Metode: Provedeno je retrospektivno istraživanje bolničke baze podataka za period od 2007. do 2017. godine. Identificirano je 46 PBC pacijenata s dostupnom medicinskom dokumentacijom. Analizirani su demografski, klinički i epidemiološki podatci, odgovor na terapiju nakon 12 mjeseci korištenja, te klinički ishod bolesti. Lošim kliničkim ishodom smatrani su: smrt, razvoj raka jetre, potreba za transplantacijom jetre ili jetrena dekompenzacija. Za procjenu incidencije i prevalencije uzeti su u obzir samo bolesnici s prebivalištem na geografskom području odgovornosti KB Dubrava. ----- Rezultati: Uključeni bolesnici bili su pretežito ženskog spola (87%) s medijanom starosti od 57,5 godina, a 68,8% ih je bilo u ranom kliničkom stadiju bolesti. Analizom bolesnika s prebivalištem na području odgovornosti KBD (N=38) izračunata je prevalencija od 11,47 bolesnika na 100,000 stanovnika i godišnja incidencija od 0,54 na 100,000 stanovnika. Svi bolesnici liječeni su ursodeoksikolnom kiselinom (UDCA). Među 32 bolesnika za koje su bili dostupni podatci nakon 12 mjeseci terapije, povoljan biokemijski odgovor zabilježen je u njih 25 (78%). Čimbenici povezani s uspjehom liječenja bili su redovito uzimanje terapije (P = 0,007) i niska inicijalna koncentracija IgM-a (P = 0,009). Loš klinički ishod zabilježen je u 5 bolesnika. U multivarijatnoj analizi osteoporoza je bila jedini čimbenik neovisno povezan s lošim ishodom bolesti (HR = 9,27; P = 0,031). ----- Zaključak: PBC je rijetka bolest, koja u opisanoj kohorti pogađa pretežito žene, srednje dobi, a većina ih ima blagi klinički stadij i dobru prognozu. Iako zemljopisno ograničeno, ovo istraživanje pruža prve epidemiološke podatke o PBC-u u Hrvatskoj i time nadopunjuje PBC kartu Europe.Introduction: Primary biliary cholangitis (PBC) is a chronic and progressive cholestatic liver disease of unknown etiology. Nowadays, PBC is considered an autoimmune disease activated by environmental factors in individuals with a genetic predisposition. Prevalence and incidence rates of PBC appear to increase worldwide. The epidemiological gradient shows a decline in prevalence from West to East and from North to South. There are no published data on incidence, prevalence or demographic characteristics of patients with primary biliary cholangitis (PBC) in Croatia. The aim of this study was to show the epidemiological and demographic characteristics of PBC patients treated at Dubrava Clinical Hospital. ----- Methods: A retrospective search of the hospital´s database was conducted. Data from 2007 to 2017 were reviewed during the study. 46 PBC patients with retrievable medical records were identified. Epidemiologic data were restricted to patients from the hospital´s catchment area. Statistical analysis involving the entire cohort included descriptive data as well as response to therapy after 12 months and a survival analysis. Risk factors for an adverse outcome were identified. An adverse outcome was defined as a composite event (ascites, variceal bleeding, encephalopathy), or HCC, liver transplantation and death. ----- Results: Most patients were female (87%) with a median age of disease onset of 57.5 years. The cohort included 38 patients (83%) from the hospital´s catchment area resulting in the prevalence of 11.47/100.000 and incidence of 0.54/100.000 inhabitants per year. From 32 patients analyzed concerning the response to therapy, 78% responded well. Adherence to therapy (P=0.007) and low IgM concentration at diagnosis (P=0.009) were associated with therapy success. An adverse outcome was reported in five patients. Among factors related to shorter event-free survival, osteoporosis remained relevant in multivariate analysis after adjusting for age (HR=9.27; P=0.031). ----- Conclusion: Despite some drawbacks, this study gives pioneer insights into the epidemiological data of PBC in the southeast corner of Europe, therefore complementing the PBC map of Europe

POSSIBILITY OF NEW BIOTECHNOLOGY ACHIEVEMENTS IN IMPROVEMENT OF MILK QUALITY

Novija biotehnološka dostignuća ušla su u sve segmente animalne proizvodnje, pa tako i u proces proizvodnje mlijeka. Interes uzgajivača je unaprijediti genetsku osnovu i tehnologiju kako bi što učinkovitije proizveli kvalitetno mlijeko i ostvarili primjeren profit. Primjenom biotehnologija otvaraju se mogućnosti pomoću kojih je moguće napraviti korak naprijed u selekciji, unaprijediti genetsku osnovu, te uz kvalitetna grla proizvesti mlijeko koje će po svome sadržaju i po kvaliteti odgovarati točno određenim zahtjevima u proizvodnji određenog tipa i vrste mliječnih proizvoda. Od biotehnoloških metoda koje se češće koriste su embrio transfer i seksirano sjeme kvalitetnih bikova. Metode se rabe u funkciji unapređenja uzgojnog programa kao i praktične selekcije (unapređenja) stada. U radu je opisana tehnologija embriotransfera, seksiranog sjemena te polimorfizama proteina mlijeka i njihov utjecaj na količinu mlijeka, mliječne masti i proteina kao i na sposobnost mlijeka za preradu.Recent biotechnology advances have entered into all segments of animal production, including the process of milk production. The breeders are interested into improving the genetic base and technology in order to produce quality milk more effectively and make a suitable profit. Applying biotechnology opens the opportunities with which it is possible to move forward in the selection process, to improve the genetic basis, and produce milk which will, due to its content and quality, match the specific requirements in the production of a certain types and sorts of dairy products. Biotechnological methods that are often used are embryo transfer and semen sexation of high-quality bulls. Methods are used in order to improve the breeding program, as well as the practical selection (promotion) of the herds. This paper describes the embryo transfer technology, semen sexation, as well as milk protein polymorphisms and their effect on milk yield, milk fat and protein, including the ability of milk processing

Natural History of Nonalcoholic Fatty Liver Disease: Implications for Clinical Practice and an Individualized Approach

Nonalcoholic fatty liver disease (NAFLD) is becoming the most prevalent liver disease worldwide, associated with epidemics of overweight and resulting metabolic syndrome (MetS). Around 20–30% of patients with NAFLD develop progressive liver fibrosis, which is the most important predictor of liver-related and overall morbidity and mortality. In contrast to classical understanding, no significant association has been demonstrated between the inflammatory component of NAFLD, i.e., nonalcoholic steatohepatitis (NASH), and the adverse clinical outcomes. Older age (>50 years) and presence of type 2 diabetes mellitus, in addition to some genetic variants, are most consistently reported indicators of increased risk of having liver fibrosis. However, critical driving force for the progression of fibrosis and risk factors for this have still not been fully elucidated. Apart from the genetic profile, gut dysbiosis, weight gain, worsening of insulin resistance, and worsening of liver steatosis represent candidate factors associated with unfavourable development of liver disease. Cardiovascular events, extrahepatic malignancies, and liver-related deaths are the leading causes of mortality in NAFLD. As patients with advanced fibrosis are under highest risk of adverse clinical outcomes, efforts should be made to recognize individuals under risk and rule out the presence of this stage of fibrosis, preferably by using simple noninvasive tools. This process should start at the primary care level by using validated biochemical tests, followed by direct serum tests for fibrosis or elastography in the remaining patients. Patients with advanced fibrosis should be referred to hepatologists for aggressive lifestyle modification and correction of the components of MetS, and cirrhotic patients should be screened for hepatocellular carcinoma and oesophageal varices

Validation of the New Diagnostic Criteria for Clinically Significant Portal Hypertension by Platelets and Elastography

Background and aims: We aimed to validate newly proposed noninvasive criteria for diagnosing clinically significant portal hypertension (CSPH) using liver stiffness measurements (LSM) by transient elastography (TE) and platelet count. ----- Methods: Diagnostic performance of these new criteria for CSPH (LSM ≥ 25 kPa to rule in and Plt ≥ 150 × 109/L + LSM ≤ 15 kPa to rule out CSPH) were retrospectively tested in an independent cohort of consecutive patients who underwent hepatic venous pressure gradient (HVPG) measurements and liver biopsy due to suspicion of compensated advanced chronic liver disease. Suspicion of cACLD was based on LSM ≥ 10 kPa by TE or results of liver imaging, without overt signs of CSPH. Patients with conditions known to affect results of LSM (ALT > 5 × ULN, liver congestion, extrahepatic biliary obstruction, infiltrative liver neoplasms) were excluded. ----- Results: Seventy six (76) patients were included: 78.9% males, mean age 62 years, 36.8% suffered from alcoholic, 30.3% nonalcoholic fatty liver disease, 14.5% chronic viral hepatitis, 30.3% were obese, 52.6% had HVPG ≥ 10 mmHg, 56.6% had platelet count ≥ 150 × 109/L. LSM ≥ 25 kPa had 88.9% specificity (95% CI 73.9-96.9) to rule in, whereas Plt ≥ 150 + LSM ≤ 15 kPa had 100% sensitivity (95% CI 91.1-100) to rule out CSPH. ----- Conclusion: By using these simple noninvasive criteria 49/76 (64.5%) patients could be classified correctly for the presence/absence of CSPH, thus obviating the need for HVPG measurements

Infection as a predictor of mortality in decompensated liver cirrhosis: exploring the relationship to severity of liver failure

Background: Infections are common in patients with liver cirrhosis and increase mortality. We explored the relationship between infection and liver dysfunction in their effects on mortality. ----- Methods: Single-center data on decompensated liver cirrhosis patients hospitalized between March 2014 and December 2017 (index period) were reviewed until death, liver transplantation or 31 December 2018. Infections were classified as community-acquired infection (CAi) or hospital/healthcare associated infection (HCAi). Child-Pugh, model for the end-stage liver disease (MELD) and chronic liver failure-organ failure (CLiF-OF) scores indicated liver (dys)function. ----- Results: We enrolled 155 patients (85% alcoholic liver disease), 65 without infection at first hospitalization, 48 with CAi and 42 with HCAi. Multidrug resistant agents were confirmed in 2/48 (4.2%) CAi and 10/42 (23.8%) HCAi patients. At first hospitalization, infection was independently associated with worse liver dysfunction and vice versa, and with higher 30-day mortality [odds ratio (OR) = 2.73, 95% confidence interval (CI) 1.07-6.94]. The association was reduced with adjustment for MELD/CLiF-OF scores, but mediation analysis detected an indirect (via liver dysfunction) association. Twenty-eight patients were repeatedly hospitalized, 11 with new HCAi. HCAi was independently associated with twice higher risk of medium-term mortality and added an additional risk to any level of liver dysfunction, considering all or patients who survived the first 30 days. In those repeatedly hospitalized, HCAi appeared independently associated with a higher probability of infection and higher MELD scores at subsequent hospitalizations. ----- Conclusion: Infection (particularly HCAi) adds mortality risk to any level of liver dysfunction in decompensated liver cirrhosis patients. Mechanisms of long(er)-term effects (in acute episode survivors) seemingly include enhanced deterioration of liver function

Assessment of clinically significant portal hypertension by two-dimensional shear wave elastography

BACKGROUND AND AIMS: To evaluate two-dimensional shear wave elastography (2DSWE) in parallel with transient elastography (TE) for diagnosing clinically significant portal hypertension (CSPH) and high-risk varices (HRV) in patients with chronic liver disease. PATIENTS AND METHODS: Consecutive patients with suspicion of compensated advanced chronic liver disease (cACLD) [liver stiffness measurement (LSM)≥10 kPa by TE, or morphological signs suggestive of cACLD on imaging], with no history of liver decompensation, underwent hepatic venous pressure gradient (HVPG) measurement, transjugular liver biopsy and esophagogastroduodenoscopy which served as the reference methods for diagnosing CSPH, cACLD and HRV. All patients underwent LSM and spleen stiffness measurements (SSM) by 2DSWE and TE. RESULTS: Seventy-six (76) patients were included (78% men, mean age 62 years, body mass index 28.3 kg/m2 , 36.8% alcoholic, 30.3% non-alcoholic fatty liver disease, 14.5% viral hepatitis). Of them, 80.3%, 69.7%, 52.6% and 22.4% had cACLD, cirrhosis, CSPH and HRV, respectively. LSM performed better than SSM in diagnosing CSPH and HRV. For CSPH, AUROCs (0.926 vs. 0.866), optimal cut-offs (20.1 vs. 20.2 kPa) and sensitivity/specificity (80.5%/94.3% vs. 77.5% /86.1%) were comparable for 2DSWE and TE. Ruling-out of CSPH by 2DSWE (LSM at cut-off with ≥90% sensitivity (13.5 kPa) and platelets ≥150x109 /L) performed comparably to TE, with 1/24 cases falsely classified as negative. For HRV, AUROCs were similar (0.875 2DSWE, 0.851 TE) with similar optimal LSM cut-offs enabling 100% sensitivity and ruling-out HRV. CONCLUSION: LSM by 2DSWE appears to perform equally well as TE for diagnosing CSPH and ruling-out HRV in compensated chronic liver disease