Modeling ciliopathies in patient-derived primary cells.

Ciliopathies comprise a group of inherited diseases caused by mutations in genes encoding proteins that localize to cilia or centrosomes. They afflict multiple organs and are one of the most frequent monogenic causes of kidney failure in adults, adolescents and children. Primary cilia play diverse roles in cell signaling, cell cycle regulation, planar cell polarity and mechanosensing. The use of patient-derived cells possessing endogenous disease causing mutations enables the study of these processes and their dysregulation in disease. Here we describe methods to cultivate patient-derived dermal fibroblast and renal epithelial cells isolated from urine. Fibroblasts are highly robust, long-lived, and easy to culture cells in which ciliary assembly can be easily induced. Similarly, the ability to acquire and culture ciliated renal epithelial cells without patient-invasive-intervention holds great potential to further our understanding of ciliopathies. In addition to monolayer cultures, we also detail the formation of three-dimensional renal-epithelial organoids-so-called tubuloids-that demonstrate epithelial-polarization and transepithelial transport activities like those seen in vivo renal-tubules. These in vitro models are powerful tools to investigate the underlying disease mechanisms of human ciliopathies that can be employed without the need for heavy-handed genetic or molecular manipulations.

The COVID-19 pandemic - related psychological distress and job burnout among Serbian pharmacy practitioners: a cross-sectional online study.

OBJECTIVE: The current COVID-19 pandemic has increased occupational stress for all healthcare providers, making job burnout one of the most common and largely unrecognized mental health issues among healthcare professionals. Besides physicians and nurses, pharmacy practitioners were "front-line" healthcare professionals with a critical and unique role in the public health crisis. Considering this, the aim of this study was to examine distress levels and the prevalence of job burnout among Serbian pharmacy practitioners in relation to the COVID-19 pandemic. SUBJECTS AND METHODS: This cross-sectional online study was conducted in April and May of 2020. A total of 176 pharmacy practitioners anonymously and voluntarily completed the two-section questionnaire, consisting of Copenhagen burnout inventory, CBI and 4DSQ Distress subscale. The two-section questionnaire was distributed online, among various social-media groups of pharmacy practitioners, as well as by personal contacts following the "snowball" sampling method. RESULTS: Results revealed moderately high burnout scores in our sample. The majority of the participants showed signs of personal-related job burnout, followed by work-related and client-related burnout. A strongly elevated distress level was obtained in almost two-thirds of study respondents. In addition, a significant and high correlation of all CBI subscales with distress was found, as well as medium correlations with sleep duration shortening as a mediator between distress and job burnout. CONCLUSIONS: Results of our study showed that job burnout significantly increased among pharmacy practitioners during the COVID-19 pandemic. Furthermore, we discovered that stress has an indirect impact on study participants' burnout via insufficient sleep.

Predictors of health-related quality of life in Serbian patients with head and neck cancer.

BACKGROUND: The aim of this study was to identify predictors of the Health-Related Quality of Life (HRQoL) in patients with head and neck cancers (HNCs). MATERIAL AND METHODS: In total, 345 patients with HNCs were interviewed. A self-report questionnaire was administered to collect data about demographic characteristics, health status, smoking, alcohol consumption habits, and HRQoL. It were used the EORTC Instruments - Quality of Life Questionnaire-Core 30-questions (QLQ-C30), Quality of Life Questionnaire - Head and Neck Module 35-questions (QLQ-H&N 35) and OHIP-14 instrument for HRQoL assessments. Clinical information and treatment data were collected from medical records. RESULTS: Five groups of HRQoL predictors were identified: demographic, socioeconomic, behavioral, psychophysical, and clinical/treatment. These HRQoL predictors had a strong (i.e., age, level of social support and social contact, level of education, depression, fatigue, presence of gastrostomy, comorbidities, and use of pain medications and supplements), a moderate (i.e., marital status, smoking, sexuality problems, time since diagnosis, presence of tracheostomy, and side effects outcomes of radio and chemotherapy) and a small impact (i.e., employment/financial difficulties, tumor site and stage, and surgical procedure). CONCLUSIONS: Study identified nineteen predictors that had significant, moderate and small impact on the HRQoL of patients with HNCs. Some of the predictors, like levels of social support and social contact, depression, and comorbidities could be targets for innervations to improve HRQoL.

Validation of the oral health impact profile - 14 in patients with head and neck cancer.

BACKGROUND: The Oral Health Impact Profile-14 (OHIP-14) was designed to assess patients' perception of the impact of oral disorders on their quality of life (QoL). Although the OHIP-14 is now frequently used in patients with head and neck cancer, data related to its measurement properties in this population are scarce. The aim of the present study was to evaluate the reliability and validity of the OHIP-14 in a sample of Serbian patients with head and neck cancer. MATERIAL AND METHODS: Data were available for 345 patients (257 [74.5%] males; aged 30-92 years), with head and neck cancer. All patients completed the OHIP-14 and the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C30 and QLQ-H&N35. Factor analyses, internal consistency reliability (i.e., Cronbach's α coefficient), and construct validity were analyzed. RESULTS: The factor analyses confirmed that 14 OHIP items were measuring a single underlying factor. Cronbach's α coefficient was 0.98 and corrected item-total correlations ranged 0.77-0.93. Lower OHIP-14 scores (i.e., lower impacts on oral health) were more frequently present among patients who had only surgery as a therapeutic procedure compared to those who had surgery accompanied with radio- and chemotherapy (p < 0.01). Patients with a tumor stage 0-II also had lower OHIP-14 scores compared to those who had a tumor stage III-IV (p < 0.01). The OHIP-14 correlated significantly with the QLQ-C30 and QLQ-H&N35 scales. CONCLUSIONS: As a unidimensional instrument, the OHIP-14 provides oral QoL assessments with sound internal consistency reliability and construct validity among patients with head and neck cancer.

Accuracy of reported food intake in a sample of 7-10 year-old children in Serbia.

OBJECTIVES: Children's ability to recall and report dietary intake is affected by age and cognitive skills. Dietary intake reporting accuracy in children is associated with age, weight status, cognitive, behavioural, social factors and dietary assessment techniques. This study analysed accuracy of 7-10 year-old children's reported food intake for one day. STUDY DESIGN: Validation study. METHODS: Sample included 94 children aged 7-10 years (median = 9 years) from two elementary schools in a local community in Serbia. 'My meals for one day' questionnaire was a combination of 24-h recall and food recognition form. It included recalls for five meals: breakfast at home; snack at home; lunch at home; snack at school and dinner at home. Parental reports were used as reference information about children's food intake for meals obtained at home and observation was used to gain reference information for school meal. Observed and reported amounts were used to calculate omission rate, intrusion rate, corresponding, over-reported and unreported amounts of energy, correspondence rate and inflation ratio. RESULTS: Overall omission rate (37.5%) was higher than overall intrusion rate (36.7%). The same food item (bread) has been the most often correctly reported and omitted food item for breakfast, lunch and dinner. Snack at school had the greatest mean correspondence rate (79.6%) and snack at home the highest mean inflation ratio (90.7%). CONCLUSION: Most errors in children's recalls were incorrectly reported amounts and not the food items. The questionnaire should be improved to facilitate accurate reports of the amounts.

Dietary influence on breast cancer.

Certain lifestyle and environmental factors play an important role on breast cancer (BC) risk, but data on the influence of nutritional factors are still conflicting. Migrational data have pointed to nutrition as one of the more relevant external factors involved. So far, the only well established nutrition-related risk factors for BC are obesity and alcohol consumption. The evidence that body fatness is a cause of postmenopausal breast cancer is convincing as is the evidence that alcoholic drinks are a cause of BC in all ages. On the other hand, body fatness probably protects against BC diagnosed premenopause. It is more likely that the BC risk is related to life-long dietary habits. The general preventive recommendation often includes a reduction of alcohol, red meat and total dietary fat, and increase in vegetable and fruit consumption. The purpose of this work was to summarize and present current opinions on the influence of diet and nutrition on BC etiology and to suggest possible preventive measures.

Controlled assembly of poly(D,L-lactide-co-glycolide)/hydroxyapatite core-shell nanospheres under ultrasonic irradiation.

An ultrasound field was applied to obtain PLGA/HAp biocomposite nanospheres. Formulation of PLGA/HAp composite revealed significant dependence of the morphology of the obtained composite on synthesis parameters, like the intensity of applied ultrasonic field, polymeric and ceramic parts' wt.% ratio in the composite, temperature of the medium, type of surfactant, and the sequence of steps in the formation of PLGA/HAp. Optimal parameters for the formation of PLGA/HAp included a lower content of the ceramic phase (PLGA/HAp=90:10), higher power of ultrasonic field (P=142.4W), lower temperature of the medium during ultrasonic treatment (T=8 degrees C), dilute solution of PVP as surfactant and dispersion of hydroxyapatite in polymer solution in order to achieve required homogeneity before the formulation of the composite. The morphology of PLGA/HAp particles synthesized under these conditions was highly regular: sphere-like, with particles of very small dimensions (150-320nm), highly uniform particle size distribution and characteristic planar spatial self-organization. These characteristics indicate significant improvements in PLGA/HAp composite resulting from ultrasonic method.

Triple-way therapeutic approach for paraganglioma-dependent erythrocytosis: drugs and surgery plus "multi-manner" apheresis.

Paragangliomas are tumors arising from the extra-adrenal paragangliar neural crest cells. The sympathoadrenal neuroendocrine system consists of extra-adrenal paragangliar cellular layer along the paravertebral and para-aortic axis, and the adrenal medullae. Paraganglioma should be included in the differential diagnosis of secondary erythrocytosis due to its possible ectopic erythropoietin (EPO) secretion. Thus, in this report we present a 24-year-old female patient with onset of unregulated ectopic EPO secretion, and consecutive erythrocytosis followed by hypertension, secondary to paraganglioma of multifocal retroperitoneal localization. Clinical, laboratory, and radiological investigations confirmed both an elevated EPO level and the presence of multiple paraganglioma. This paraneoplastic-mediated medical condition with high risk of cellular hyperviscosity syndrome (CHVS) requires prompt diagnosis and rapid therapeutic interventions. Initially, simple phlebotomy procedures were used; following that, tumors were surgically removed. In the course of the disease, this tumor relapsed, and urgent apheresis, as a treatment of life-threatening state, was used. The therapy performed resulted in a rapid blood viscosity depletion and a significant (P < 0.01) serum EPO reduction, as well as the general clinical benefit. Therefore, we conclude that the use of our own "multi-manner" apheresis (erythrocythapheresis plus plasma exchange), for long-time interval (until further causative therapy), effectively cross-bridged the possible hazards of EPO-dependent CHVS.

[Importance of studying exposure of the population to radon and its daughters]. / Znacaj istrazivanja izlozenosti stanovnistva radonu i njegovim potomcima.

INTRODUCTION: Radon and thoron are naturally-occurring radioactive gases, which are products of uranium and thorium decay series, respectively. Uranium and thorium occur widely in the environment, in rocks, soil, air, water, building materials, humans, etc. Radon daughters in the air are predominantly attached to aerosols. A minor part, normally less than 10%, occurs as unattached atoms or ions. The relative distribution of attached daughters in indoor air and equilibrium factor depend on many variables, such as the decay constant, the concentration and size distribution of aerosols and ventilation rates. Increased ventilation decreases the concentration of radon and daughters in the air. OCCURRENCE: Concentration of radon, thoron and their decay products in the air indoors, in mines or houses, is higher than outdoors. In houses, the level of radon daughters may be enhanced by radon from radium rich building materials, landfill, soil and bedrock under the house, radon rich water and by poor ventilation. CONCLUSION: In recent years, several evaluations of human health risks and estimations have been made in regard to the dose-response relationship and lung cancer risk attributable to inhaled radon daughters. Inhalation of radon and thoron daughters leads to deposition in the human respiratory tract and consequent irradiation. Deposition depends on various factors, such as the size distribution of aerosols to which the daughter products of radon are attached, and fraction of unattached daughters. On average, the dose to the basal cell layer in the lung is about 5 to 8 times higher than the dose in the pulmonary region.

[Lipedema complicated with lymphedema and chyloderma]. / Lipedem komplikovan limfedemom i hilodermijom.

Lipedema never reveals clinical picture of extreme lymphedema-elephantiasis, and skin signs and complications have not been observed. Aim of this paper is to present a case of lipedema with the initial lymphedema in which, after one episode of lymphangiitis and cellulitis, came to the rapid development of lymphedema followed by chyloderma. During the local treatment of extreme chyloderma with excessive exudation, semiocclusive synthetic dressings have been used for moist wound healing. The treatment was completed after 20 weeks with total epithelizsation, without maceration and irritation, without additional spreading of the chyloderma field, without wound infections, with fast and full relief of the pain. Lipedem with extreme lymphedema can be followed by skin complications of lymphedema like chylodermia.

Emergency surgery of acute traumatic arteriovenous fistulas.

The purpose of this report was to analyze the clinical presentation, diagnosis, surgical and non-surgical treatment and the outcome of patients with acute AVFs seen during the last 8years. During 1991-98 we treated 47 patients with traumatic AVFs. Among these patients 11 were classified as 'acute' AVF. All the patients were male and they ranged in age from 17 to 64yr (mean 25yr). The time from injury to admission to our two institutions varied from 4h to 17days. Emergency surgery was performed in unstable patients and in those with an expanding haematoma base on clinical assessment alone. The acute surgery group consisted of stable patients that were operated after angiography examination. A ??? murmur and thrill were present in seven patients, and peripheral pulses were absent in four patients. The main blood vessels were reconstructed using various techniques in eight patients and minor blood vessel were ligated in three patients. Two lower limb amputations had to be performed, both after reconstruction of popliteal AVFs. In the remaining nine patients reconstruction of the axial vessels was successful and no signs of ischemia developed in patients after ligation of minor vessels. No neurological deficit developed in two patients whose AVFs were vaporised in the neck. The average hospital stay was 8. 6days. Emergency surgery is safe in unstable patients with traumatic AVFs. A thrill and murmur are characteristic signs even in the acute setting, but the examiner must be persistent in looking for them whenever there is a suspicion of a blood vessel injury. Angiography is a reliable diagnostic tool in stable patients, but whether it is essential when there is an expanding pulsating hematoma remains debatable.

Blast injury from explosive munitions.

OBJECTIVE: To evaluate the effect of blast in common war injuries. METHODS: One thousand three hundred and three patients injured by explosive munitions and demonstrating extremity wounds without other penetrating injuries were admitted to the Military Medical Academy in Belgrade between 1991 and 1994. Of these, 665 patients (51%) had symptoms and physical signs that were compatible with the clinical diagnosis of primary blast injury, whereas the remaining 658 patients did not. RESULTS: Random sampling of 65 patients in the blast group during the early posttraumatic period showed statistically significant elevations in blood thromboxane A2 (TxA2), prostacyclin (PGI2), and sulfidopeptide leukotrienes compared with the random sample of 62 patients in the nonblast group. This difference could not be accounted for by differing injury severity between the groups, because the severity of wounds as measured by both the Injury Severity Score and the Red Cross Wound Classification was similar in both groups. Amongst blast patients, 200 patients (30%) had long-term (1 year) symptoms and signs reflecting central nervous system disorders. These symptoms and signs were only sporadically found in 4% of the nonblast patients. These findings indicate that primary blast injury is more common in war injuries than previously thought and that of those affected by blast, a surprisingly high proportion retain long-term neurologic disability. The elevation in eicosanoids could be used to confirm and monitor blast injury. CONCLUSION: In relation to the immediate management of patients injured by explosive weapons, it follows that particular attention should be paid to the presence and/or development of blast injury. Our findings indicate that blast is more common in war injuries than previously thought. Eicosanoid changes after blast injury suggest that blast injury causes a major physiologic stress. A variety of effects on the central nervous system suggest that blast injury could be responsible for some aspects of what is now considered to be the posttraumatic stress disorder.

[Management of war injuries of blood vessels in prolonged ischemia]. / Zbrinjavanje ratne povrede krvnih sudova kod produzene ishemije.

We present a successful management of a gunshot wound of major blood vessels of the leg, in condition of prolonged ischemia. The patient has been sent to our hospital for the complete vascular management of the complete interruption of the major blood vessels of the left leg that were primarily ligated and marked. Because of the difficulties in evacuation, the patient arrived 20 hours after the wounding. There was no indication for the arteriography. The interruptions of the continuity of the superficial femoral artery in length of approximately 10 cm and of deep femoral vein in length of 8 cm were managed by interposition of autovenous grafts taken from the large subcutaneous vein of the other leg. Standard fasciotomy and partial exclusion of muscles of the left lower leg with relaxing fasciotomy of the left upper leg muscles were performed in the operative procedure. A successful rehabilitation has been completed and five years after the wounding, completely correct vascular and functional condition of the wounded leg was observed.

[Contusion injury of the brachial artery]. / Kontuziona povreda brahijalne arterije.

We present a case of a successful repair of brachial artery after blunt trauma, almost five days after the injury. Autovenous graft (great saphenous vein) was used for the repair. The diagnosis was confirmed by angiography, and this method was indisposable in the postoperative course. Precise surgical technique is compulsory in obtaining optimal surgical revascularization.

[Complications after primary surgical management of war injuries of the colon and rectum]. / Komplikacije posle primarnog hirurskog zbrinjavanja ratnih povreda kolona i rektuma.

War injuries of colon and rectum are real challenge for war surgeons. The aim of the study was to point out the necessity of war surgical doctrine application, considering the risk factors in the choice of primary surgical procedure and surgeon's experience in managing this kind of injuries. Postoperative complications and indications for reoperations in 216 wounded after primary surgical management of colonic and rectal war injuries in the period 1991-1995 were retrospectively analyzed. A total of 183 wounded (84.7%) were primarily surgically managed in war hospitals. Combined injuries of colon and/or rectum or more than 2 abdominal organs were found in 197 wounded (91.2%), and 29.1% of wounded were also with the combined extra-abdominal injuries. Postoperative complications were observed in 58 wounded (26.8%) and were associated with the injury and primary management of colon and rectum, with septic complications in 87.9% of cases. Reoperation was performed in 44 wounded (75.8%) and the most frequent indications, such as diffuse peritonitis and stercoral fistula, were the sequlae of dehiscence of suture or anastomosis, overlooked lesions and secondary perforations (blast) of colon or rectum. Principle of complete diversion of stool/intestinal content was used in reoperation. Mortality rate in reoperated group was 20.4% (9 wounded). Cause of death was diffuse peritonitis followed by consecutive sepsis and MOFS. The significance of surgical war doctrine and risk factors in the choice of primary surgical procedures were emphasized.

[The Miles-Thompson-Bricker method of total pelvic exenteration in malignant tumors of the rectum with urinary bladder infiltration]. / Totalna pelvicka egzenteracija po Miles-Thompson-Brickeru kod malignih tumora rektuma koji infiltruju mokracnu besiku.

The radical method of surgical treatment of malignant rectal neoplasms, total pelvic exenteration by Miles-Thompson-Bricker, was introduced in the Clinic for General and Vascular Surgery of Military Medical Academy in October, 1993, 45 years after the first operation of that kind in the world. This method of surgical treatment was indicated in 3% of our patients with malignant neoplasms of the distal part of rectum that infiltrated the urinary bladder. The life of patients with two stomas though not easy, was prolonged.

Glucose as an adjunct triage tool to the Red Cross Wound Classification.

The plasma concentrations of glucose, adrenaline, noradrenaline, insulin, and cortisol were measured in 59 patients within 18 hours of military gunshot/missile (MG/M) wound. The wounds were categorized by the Red Cross Wound Classification (RCWC) and assessed by the Injury Severity Score (ISS) method. The majority of the measured biochemical parameters, except insulin, were significantly increased after MG/M wounds, compared with control values. Plasma glucose concentration in wounded patients was positively related to ISS over the whole severity range. Plasma insulin concentration increased with glucose. Noradrenaline and cortisol were positively related to glucose. Because hemorrhage is the most common cause of general response to MG/M wound, we concluded that glucose measurement could be a useful adjunct tool to the RCWC in rapid and accurate assessment of severely wounded patients, especially those with occult thoraco-abdominal wounds.

Treatment of wounded in the combat zone.

Authors presented their own experiences in treating 735 wounded in high-intensity combat zones in the territories of former Yugoslavia during 1991 to 1992. The mobile field hospital with surgical crews was situated 5 to 10 km from the front line, and its basic task had been continuous triage, immediate resuscitation with vital surgical aid, as well as organization of adequate primary and secondary air evacuation. At the field hospital level, fresh wounds were explored according to principles of war surgery, and major surgical interventions were performed in 3.3% of the wounded. Patients with massive hematothorax were treated with autotransfusion. Mortality at this primary level, field hospital was 0.75% with primary immediate resuscitation and 1.9% with immediate evacuation. We concluded that immediate resuscitation with delayed transport had advantages, compared with fast evacuation of only the wounded.

Colon and rectal war injuries.

The results of retrospective analysis in the treatment of 189 wounded with colorectal lesions treated at the Military Medical Academy from July 1991 to December 1993 were presented. Primary surgical management was performed in 33 (17.5%) wounded. The others were transported into this hospital for further treatment after primary surgical management in war hospitals in the combat zone. Colorectal lesions (46.0%) were primarily inflicted by bullets of various calibers and high kinetic energy. The time interval between wounding and surgery was less than 6 hours for 39.6% of the wounded, whereas 80.1% were operated on within 12 hours. Multiple or combined colonic and rectal injuries were found in 91.5% of the wounded. Different surgical procedures were performed in accordance with the surgical war doctrine. Postoperative complications (colorectal cause) were found in 40 (21.2%) wounded. Reoperation was performed in 35 (18.5%) wounded. Total mortality rate was 10.1% (19 wounded).