Introducing in vivo pancreatic cancer models for the study of new therapeutic regimens

Introduction: Pancreatic cancer is a severe oncological disease with an ever-increasing incidence and a high rate of morbidity and mortality. Therapeutic options are limited and the five-year overall survival rate is 7­20%, depending on the possibility of surgical resection and the earliness of detection. Most patients with this diagnosis die due to the resistance of tumour cells and their microenvironment to the used treatment regimes. Methods: In our study, we focused on the implementation of two in vivo models, which are the cell-line derived xenograft (CDX) and the patient-derived xenograft (PDX). These two models differ significantly from each other methodologically, technically, financially, but also in their achieved results. Results: In a pilot study, we managed to successfully implement the CDX model with a very aggressive and resistant PaCa-44 line of pancreatic cancer in a total of 30 NU/NU strain mice. Furthermore, we created three PDX models with various subtypes of pancreatic cancer from patients operated at the University Hospital Kralovske Vinohrady, Department of General Surgery. These tumours were re-transplanted into subsequent generations of 23 individuals of NOD/SCID strain and 47 NU/NU strain mice. The established CDX and PDX models are then used to compare conventional and experimental chemotherapy regimens. Conclusion: The next steps will be to evaluate the effects of treatment regimens by using imaging and molecular genetic methods and to optimise the entire process for further use in precise personalised medicine for patients with pancreatic cancer. The upcoming goal is to create a library of PDX models of the most common pancreatic ductal adenocarcinoma and other rare subtypes of pancreatic cancer.

T cells and their function in the immune response to viruses.

The development of CD4+ T helper cells is determined by the set of transcription factors and the genes these transcription factors transcribe. In this review, we describe the basic nature of Th1, Th2, Th9, Th17, T-follicular helper (Tfh), gamma delta (γδ) T cells, and T-regulatory (Treg) cells subsets, their master regulator transcription factors and their corresponding signature cytokine production profiles. Cellular immunity plays important role during virus infection. Optimal immune response to viral infections require a gentle balance of effector responses to clear the infected cells and regulatory mechanism to prevent  immunopathology. The behavior of the helper cells differs with each virus - while in some cases, the response is beneficial; in other cases, it is harmful. We discuss the protective and pathological role of T cell immunity against influenza A virus (IAV), respiratory syncytial virus (RSV), immunodeficiency virus type 1 (HIV-1), and hepatitis B virus (HBV) infection. Keywords: T cell; cytokine; influenza virus; respiratory syncytial virus; hepatitis B virus; human immunodeficiency virus type 1.

[Successful treatment of azoospermia diagnosed as "Sertoli cells only syndrome with maturation arrest"--developing a quality embryo]. / Uspesné resení azoospermie s diagnózou "Sertolli cells only syndrome" (SCO sy.) a Maturation Arrest (MA)--získání kvalitního embrya.

OBJECTIVE: The transfer of good quality embryo in the program of assisted reproduction in the case of azoospermia, dg. Sertolli cells only syndrome (SCO sy) + maturation arrest (MA). Testes were assessed and found to have a high occurrence of Sertolli cells and very low occurrence of germinal cells, which were arrested at the round spermatid level. The histological evaluation was hypospermatogenesis gr. 3 (minimum 1 spermatid/sample). DESIGN: Case report. SETTING: Laboratory IVF, Iscare, a. s., Department of Biology and Biochemistry of Fertilization, Institute of Molecular Genetics, Czech Academy of Sciences, Prague. SUBJECT AND METHOD: The successful integration of three methods provides a solution for this case of azoospermia. Immunology and histology can more exactly diagnose the degree of azoospermia. Detection and visualisation of spermatids using monoclonal antibodies against sperm proteins predicts the eventual occurrence of spermatogenesis, and histological evaluation confirms these immunological findings. Using the information of both methods it is possible to use special in vitro cultivation of testicular cells and so obtain injectable spermatozoa, or precursors of sperm, for the ICSI method. CONCLUSION: The probability of acquisition of good-quality embryo in the program of assisted reproduction is higher when these three methods are applied in combination.

[Surgical treatment of male infertility]. / Taktika chirurgické lécby muzské infertility.

OBJECTIVE: In the Czech Republic in the treatment of male infertility the most up-to-date technologies of assisted reproduction have also become the methods of choice: MESA--microsurgical epididymal sperm aspiration, TESE--testicular sperm extraction and ICSI--intracytoplasmic sperm injection. The objective of the present study was an attempt to find a reliable predictor of results of planned TESE as prediction of possible successful sperm extraction can prevent unnecessary ovarian stimulation. DESIGN: Retrospective analysis of a group of 202 men operated in the authors centre on account of azoospermia--65 patients with obstructive azoospermia and 137 patients with testicular azoospermia. SETTING: Andromeda, Biolab, Iscare IVF, Prague. METHOD: The following parameters were evaluated: testicular volume, FSH serum level and histological finding and their values were compared with results of TESE. RESULTS: Elevated FSH levels and low testicular volume do not rule out a positive result of TESE. The most accurate predictor is histological examination. Elongated spermatids were found in the histological material in 59.3% of the examined cases which correlates significantly with the 60.2% TESE yield in the evaluated group. CONCLUSION: The material assembled on operation was divided into three parts. One serves preparation of the perIerative native preparation, the second one is dispatched for histological examination and the largest part is frozen. Simultaneous collection of oocytes and sperm cells is done only in selected cases. The histological finding of elongated spermatids indicates a positive finding of sperm during TESE with an accuracy of 91%. Assessment of the causes of azoospermia, evaluation of the state of spermatogenesis and treatment are thus implemented by a single operation.

[MESA, TESA, TESE + ICSI: results of the first 50 cases]. / MESA, TESA, TESE + ICSI: výsledky u prvních 50 prípadu.

During the period between February 1996 and August 1997 51 infertile men were operated. The authors evaluate the results from 47 couples--25 cases of testicular azoospermia and 22 cases of obstructive azoospermia. The mean age of the men in the group was 33 years (22-48 years). The follow-up period is 1-18 months. In 17 cases, microsurgical epididymal sperm aspiration (MESA) was made, in five cases testicular sperm aspiration (TESA) and in 25 men, testicular sperm extraction (TESE) was performed. In the group with testicular azoospermia it proved possible to obtain sperm in 12 of 25 cases, i.e. in 48%. In men with obstructive azoospermia all aspirations were successful, i.e. the yield was 100%. In this group five children were born, seven pregnancies are under way. Thus regardless of the etiology of male infertility 12 of 47 cases, i.e. 25.5% were resolved successfully. When the results of the two groups are differentiated, the outcome is as follows: in the group with testicular azoospermia one child was born and two pregnancies are under way, i.e. 12%, in the group with obstructive azoospermia four children were born and five pregnancies are under way, i.e. 40.9%.

[Microsurgery and micromanipulation in the treatment of male infertility: results of a one-year investigation]. / Mikrochirurgie a mikromanipulace v lécbe muzské infertility: výsledky rocního sledování.

A one-year study of 51 infertile couples, 47 couples evaluated--25 cases of testicular azoospermia and 22 cases of obstructive azoospermia. The mean age of the men in the group is 33 years (22-48 years). The follow-up period is 1-18 months. In 17 instances microsurgical epididymal sperm aspiration--MESA was made, five times testicular sperm aspiration--TESA and in 25 men testicular sperm extraction--TESE was used. In the group with testicular azoospermia it proved possible to obtain sperm in 12/25 cases, i.e. in 48%. In men with obstructive azoospermia all aspirations were successful, i.e. the yield was 100%. In this group five infants were born, another seven pregnancies are under way. Thus regardless of the etiology of male infertility 12/47 cases, i.e. 25.5%, were successfully resolved. When using differentiated evaluation of the two groups the results are as follows: in the group with testicular azoospermia one infant were born and five pregnancies are under way, i.e. 40.9%.

[The effect of extracorporeal shock-wave lithotripsy on renal function]. / Vliv litotrypse extrakorporálními rázovými vlnami na renální funkce.

Using the method of phase scintigraphy, the authors examined 48 patients who were treated by means of extracorporeal shock-wave lithotripsy. The authors proved at the 5% of significance that the development or deterioration of renal functions does not depend on the localization of the concrement and the number of shock-waves needed for disintegration. The authors confirmed the significant effect of the size of concrement on the lesion of renal function--all stones larger than 30 mm lead in 100% to a varying deviation, as compared with the examination made before extracorporeal shock-wave lithotripsy. Obesity has also a significant effect on the development or deterioration of lesions of renal function. From the total number of patients where after lithotripsy a lesion of renal function developed or deteriorated 70.6% were obese! After 24 months in two patients deterioration of the condition, as compared with the initial state, was proved.

[Surgical treatment of obstructive azoospermia]. / Chirurgická lécba obstrukcní azoospermie.

In 23 infertile men the diagnosis of azoospermia was established. Bilateral biopsy of the testes was made, in four patients primary deficiency of spermiogenesis was revealed, in another four patients the Sertoli cells only syndrome. In a group of 3 men with hypospermiogenesis revision revealed severe hypoplasia of the epididymis which in context with the bioptic finding did not give hope of successful surgery. In the group of four men with normal spermiogenesis revision revealed in one obliteration of the terminal portion of the d. deferens, treated by transurethral discision of the colliculus and in three its partial or total agenesis. In one of the three latter cases bilaterally artificial spermatoceles were implanted which function satisfactorily; in the aspirated contents, however, even after several months only non-motile sperm cells were found. In seven patients epididymovasoanastomosis was performed by the microscopic technique--five times unilateral and twice bilateral. In one patient a condition after bilateral vasectomy was involved which was resolved by vasovasoanastomosis. In three patients after epididymovasoanastomosis azoospermia persists. In another two patients the check-up spermiogram revealed a positive finding, the wife of one of the patients is pregnant at the moment. In one patient the result of the operation cannot be evaluated yet due to a short time interval after operation. One patient did not come for a check-up examination. The patient after vasovasoanastomosis has a positive spermiogram with 38 million sperm cells per ml. In one patient with a discision of the colliculus the spermiogram is negative. The cumulative index of patient anastomoses reported in the literature is approximately 45%, the percentage of pregnancies only 17-18%.

[Problems with indications for extracorporeal lithotripsy in urolithiasis]. / Indikacní problémy extrakorporální litotrypse urolitiázy.

Extracorporeal lithotripsy is a modern noninvasive method for the treatment of urolithiasis. The morphological and functional state of the urinary pathways, the localization of concrement and their size and number call in 35% of the patients for additive and auxiliary operations. Such comprehensive treatment can be provided by departments where all methods for the treatment of lithiasis are readily available. Repeated lithotripsy and combined therapeutic methods of more complicated conditions protract hospitalization and reduce the number of patients. The authors tested in more than 2500 lithotripsies the possibility to apply this treatment in the great majority of lithiases. However, because of the limited capacity they had to reduce the spectrum of indications to a minimum. Retardation of treatment by a protracted waiting period could damage the patient and it is then better to indicate another therapeutic approach; or to provide adequate treatment. We hope that patients who will need treatment after a year's time, will have the opportunity to be treated sooner by lithotriptors which should be installed before long in other departments.

[Nephrolithotomy using Doppler and dynamic sonography]. / Nefrolitotomie pod kontrolou dopplerovské a dynamické sonografie.

In open operations of coralliform or multiple small nephrolithiasis frequently the problem of peroperative localization of residual concrements arises. A simple, reliable, accurate and non-aggressive peroperative detection of concrements in the kidney is made possible by ultrasonography. The method was used in 25 cases, a probe with a frequency of 7.5 MHz makes it possible to detect even minute fragments. On the screen we can see not only the size of concrements but also their distance from the surface, and their position can be accurately assessed three-dimensionally. The surgeon thus can make nephrotomy aimed directly on the concrement. The course of major blood vessels can be detected by means of a Dopplers probe. This procedure makes it possible to make the incision in a relatively avascular zone without the necessity of the application of cooling or ischaemia of the kidney. In the investigated group no false positive finding was obtained, a false negative finding can result from non-systematic examination.

[Acceleration of the chemolysis of kidney stones using ultrasound. Experimental study in vitro]. / Urychlení chemolýzy ledvinových kamenu pusobením ultrazvuku. Experimentální studie in vitro.

The dissolution of infectious renal concrements by means of percutaneous nephrostomy or ureteral catheter often requires lavage lasting several weeks. Using an experiment in vitro, the authors tried to reduce this interval by means of ultrasound. By combination of the physical effect of ultrasound with that of chemolytic dissolution of the infectious concrement was significantly accelerated. Thus, simultaneous intermittent ultrasound application to the body's surface during dissolution could considerably reduce the time needed for the concrement to dissolve in vivo.

[Extracorporeal shock-wave lithotripsy]. / Litotrypse extrakorporálními rázovými vlnami.

Concrement lithotripsy in the kidney and ureter using extracorporeal shock waves (ESWL) was performed with the aid of a 2nd-generation lithotriptor--Lithostar Siemens. Over a period of 11 months ESWL was applied in 526 patients aged 7 to 80 years, 10 of these were children. 19 patients had bilateral lithiasis so that the treatment was applied to a total of 545 renoureteral units and 698 concrements were disintegrated. In the ureter 34 concrements were disintegrated, the remainder being in the kidney. 12 patients had solitary kidney lithiasis; 62 subjects prior to ESWL, had been operated on the same kidney for lithiasis or on the ipsilateral ureter. Most patients had only local infiltration anaesthesia of the relevant intercostal nerves sometimes supplemented with mild opioid analgesia. Children were operated on in general anaesthesia. More than one ESWL application was needed in 33% cases, 54 disintegrated concrements measured more than 30 mm. Cystine concrements in 2 patients remained unaffected, in 1 woman with apatite lithiasis the disintegration was unsatisfactory. Auxiliary interventions had to be performed in 22% cases, prior to ESWL, and in 13% patients after ESWL, in 1 case following ESWL, open ureteromy was performed. The patient's exposure to radiation is negligibly low, the paramedical personnel is exposed to none. The noise level both for the patient and personnel is within the hygienic limits and is lower than in the case of 1st--generation lithotriptors. The average hospitalization period in patients with unilateral lithiasis lasted 7 days, 8 patients were treated in out-patient departments. When released from the clinic, 39.5% patients had no concrements, the remainder had only minute residual concrements. Three months upon the release, 77.6% patients were found with no demonstrable signs of concrements.(ABSTRACT TRUNCATED AT 250 WORDS)