Human enteric neurons: morphological, electrophysiological, and neurochemical identification

BACKGROUND: Access to tissue, difficulties with dissection, and poor visibility of enteric ganglia have hampered electrophysiological recordings of human enteric neurons. Here, we report a method to combine intracellular recording with simultaneous morphological identification of neurons in the intact myenteric plexus of human colon ex vivo. METHODS: Specimens of human colon were dissected into flat-sheet preparations with the myenteric plexus exposed. Myenteric neurons were impaled with conventional microelectrodes containing 5% 5,6-carboxyfluorescein in 20 mM Tris buffer and 1 M KC. KEY RESULTS: Electrophysiological recordings identified myenteric neurons with S and AH type properties (n = 13, N = 7) which were dye filled and classified during the recording as Dogiel type I (n = 10), Dogiel type II (n = 2), or filamentous (n = 1) cells. This classification was confirmed after fixation, in combination with immunohistochemical characterization. CONCLUSIONS & INFERENCES: This method allows electrophysiological characterization with simultaneous identification of morphology. It can be used to identify recorded cells immediately after impalement and greatly facilitates recordings of human myenteric neurons in freshly dissected specimens of tissue. It can also be combined with immunohistochemical labeling of recorded cells

Effect of lipid parameters on fetal growth in type 2 diabetes mellitus and gestational diabetes mellitus pregnancies

Background: During pregnancy,complex changes occur in lipid profiles. The aim of the study was to evaluate the effect of lipid parameters on fetal growth in type 2 diabetes mellitus (D.M) and gestational diabetes mellitus (GDM) pregnancies. Material and methods: In forty three type 2D.M. and two hundred GDM women pregnancies were analyzed: age, body mass index (BMI), lipid parameters, HbA1c in first, second and third trimester of pregnancy, preeclampsia, and baby birth weight. Results: D.M. tip 2 and GDM group were statistically significant different in the following variables: total lipids, triglycerides, total cholesterol, BMI, age, baby birth weight, and incidence of SGA (9.4±2,3 vs. 11,0±2,3mmol/L, 2,4±1,4 vs. 3,4±1,6mmol/L, 5,5±1,2 vs. 6,4±1,4mmol/L, 30,6±5,4 vs. 26,9±5,2 kg/m2, 34±7,8 vs. 31,5±5,6 years, 3183±972 vs. 3533±699 g., 20% vs. 7,5%, respectively, p<0,05). Statistically significant correlations were found between triglycerides and HbA1c (r=0,18, p<0,05), HDL-C and HbA1c (r=-0,19, p<0,05), HDL-C and large for gestational age (LGA) (r=-0,17, p<0,05), small for gestational age (SGA) and Hba1c (r=0,29, p<0,05). LinearmultipleregressionanalysisdemonstratedthatLDL-C,triglycerides, and total cholesterol were independent predictors of LGA (p<0,05). Conclusion:LDL-C and triglycerides are predictors for macrosomia in type 2 D.M and GDM pregnancies.Thus, with good regulation of lipid profile we can avoid macrosomia from type 2 D.M and GDM pregnancies. Key words: lipid parameters, gestational diabetes mellitus, type 2 diabetes mellitus, macrosomia

Евалуација на педијатриски пациенти со јувенилен идиопатски артрит третирани со биолошка терапија Тоцилизумаб (Actemra)

Juvenile idiopathic arthritis (JIA) is the most common chronic disease in childhood. It manifests a heterogenic group of symptoms of arthritis, lasting at least 6 weeks and it appears before the age of 16. Patients who had no good therapeutic response to conventional therapy with Methotrexate were treated with biological therapy. The aim of this paper was to evaluate 9 patients who were receiving Tocilizumab at the Department of Rheumocardiology, University Clinic of Pediatric Diseases in Skopje. Materials and methods: Our study included 9 patients treated at our Department with biological therapy with Tocilizumab. Prior to initiation of the biological therapy, all patients underwent laboratory investigations, purified protein derivative (PPD) skin test for tuberculosis, X ray of the lungs and heart, and analysis of hepatitis markers. All patients were treated with amp. Actemra (tocilizumab) 8 mg/kg/tt i.v. Two of the patients had a severe form of the disease (one with severe systemic form and one with severe oligoarticular form of JIA). All presented patients had clinical remission of the disease. Conclusion: Therapy with tocilizumab in patients with juvenile idiopathic arthritis is a good therapeutic choice. The results obtained in our study have shown a significant therapeutic effect of tocilizumab even in severe forms of the disease. &nbsp;Јувенилниот идиопатски артрит е најчеста форма на хронично заболување во детската возраст, кое се манифестира со&nbsp; хетерогена група на симптоми на артрит коишто траат најмалку 6 недели, а се појавиле пред 16-годишна возраст. Пациентите кои не дадоа добар тераписки одговор на конвенционалната терапија со Метотрексат, беа префрлени на биолошка терапија. Целта на оваа студија беше да изврши евалуација на 9 пациенти кои примаа тоцилизумаб на Одделот за реумокардиологија на Универзитетската клиника за детски болести во Скопје. Материјали и методи: Во нашата студија&nbsp; беа вклучени 9 пациенти третирани на нашиот Оддел со Тоцилизумаб во период од 2010-2020 година. Кај сите пациенти пред започнување на терапијата беа направени лабораториски иследувања, прочистен протеински деривативен кожен тест за туберкулоза (ППД), РДГ на градите, и маркери за хепатит. Сите пациенти беа третирани со амп. Актемра 8мг/кг/тт и.в. Два пациента имаа тешка клиничка презентација на јувенилен идиопатски артрит (првиот со тешка системска форма на болеста, а вториот со тешка олигоартикуларна форма). Кај сите третирани пациенти настана ремисија на клиничката слика по спроведување на терапијата. Заклучок: Биолошката терапија со тоцилизумаб кај пациентите со јувенилен идиопатски артрит е добар тераписки избор. Резултатите покажаа сигнификантен тераписки ефект дури и кај потешките облици на болеста

The Connection of the Level of Estradiol in Serum and Obesity with the Endometrial Bleeding in Postmenopausal Women

BACKGROUND: Postmenopausis is a period that begins one year after the last menstrual period. Abnormal uterine bleeding could be of different origins. AIM: This study aimed to determine the association of serum estrogen hormone levels and obesity with the occurrence of endometrial bleeding in post-menopausal women. MATERIAL AND METHODS: Prospective clinical study involving 120 postmenopausal patients treated at the University Clinic for Gynecology and Obstetrics-Skopje, divided into two groups: control and study. The control group consisted of 40 postmenopausal patients without endometrial bleeding, hospitalised and operated due to urogenital pathology. The study group consisted of 80 patients with endometrial bleeding who were divided into three subgroups according to the thickness of the endometrium: from 5-8 mm, 8-11 mm and above 11 mm. In all subjects, estradiol and BMI was determined. RESULTS: Estradiol levels were statistically higher in the study group compared to control while statistically significant difference among the three subgroups according to the thickness of the endometrium about the levels of estradiol in blood is not found. About BMI, the results showed that there was no statistical significance between the two examined groups. CONCLUSION: Patients with endometrial bleeding have increased levels of estradiol and are at increased risk of endometrial cancer about controls, the likelihood of endometrial cancer significantly increases by 1,108 times

Cor triatriatum sinister - ретка вродена срцева мана- наши искуства во дијагнозата и третманот во детска возраст

Cor triatriatum sinister is a rare congenital heart defect in which left atrium is divided in two parts with the fibromuscular membrane. The proximal atrium accepts the pulmonary veins with their blood flow, the distal or (real) atrium is usually empty and separated from the ventricle by the mitral valve. There is a constant communication between the two parts of the divided atrium by which the blood flow is redirected to the left ventricle. This defect is rare, usually isolated, but it might be combined with other congenital heart defects. We present cases of two children aged 8 and 3 years at the time of establishment the diagnosis. In both cases the main clinical signs were fatigue and heart murmur on auscultation. Anamnesis, clinical symptoms and signs, ECG, chest X ray and the gold standard – echocardiography were used for establishing the diagnosis. Definitive treatment was made with cardiosurgical resection of the fibromuscular membrane using extracorporeal circulation. Regular periodical evaluation is necessary in order to discover late complications of cardiac rhythm disturbancesCor triatriatum sinister е ретка срцева мана кај која левата преткомора е поделена на два дела со фибромускулна мембрана. Проксималната преткомора ги прима пулмоналните вени заедно со крвта која тие ја носат, а дисталната или (вистинска) преткомора е вообичаено празна и таа е поделена од комората со митралната валвула. Има постојана комуникација помеѓу двата дела на поделената преткомора преку која крвта се пренасочува кон левата комора. Маната е многу ретка, најчесто е изолирана, но може да оди во комбинација со други срцеви мани. Презентираме случаи на две деца на возраст од 8 и 3 години во времето кога е поставена дијагнозата. Кај обете причина за дијагнозата бил замор и чуен срцев шум. За поставување на дијагнозата беа користени анамнезата, клиничките знаци и симптоми, ЕКГ, рендген на срце и бели дробови и златниот стандард - ехокардиографијата. Дефинитивно излекување беше направено со изведување на кардиохируршка ресекција на фибромускулната мембрана со користење на екстракорпорална циркулација. Потребно е регуларно периодично следење на децата со цел откривање на доцни компликации од типот на нарушувања на срцевиот&nbsp; ритам

Applicability of the 'Watershed Habitat Evaluation and Stream Integrity Protocol' (WHEBIP) in assessment of the stream integrity in Bregalnica River Basin

This paper reports the assessment of ecological integrity of streams in Bregalnica River Basin with emphasis on river Bregalnica as the biggest and most important watercourse in Eastern Macedonia. The results have principally been derived from remote sensing data and set up in a model build up on Watershed Habitat Evaluation and Stream Integrity Protocol (WHEBIP). WHEBIP effectiveness in predicting ecological integrity of streams has been assessed by correlation analyses derived upon data on macroinvertebrate biotic indices and physico-chemical parameters on 35 localities throughout the basin. The statistical analyses confirmed the capacity of WHEBIP to predict stream site-specific features with great accuracy in case of Bregalnica. The results obtained in this study contribute towards improvement of the WHEBIP protocol and in general promotes applicability of stream integrity assessment tools in setting priorities for integrated watershed management

The comparative anti-cancer effects of krill oil and oxaliplatin in an orthotopic mouse model of colorectal cancer

Background: Our in vitro studies demonstrated that krill oil (KO) has anti-cancer potential. This study aimed to compare the anti-cancer effects of KO with a commonly used chemotherapeutic drug, oxaliplatin and to identify the molecular mechanisms associated with KO supplementation in a mouse model of colorectal cancer (CRC). Methods: Thirty-six male Balb/c mice were randomly divided into six groups. Five groups received standard chow diet supplemented with KO (150 g/kg)), corn oil (150 g/kg), KO combined with ½ dose of oxaliplatin (1.5 mg/kg body weight/3 times per week), corn oil combined with ½ dose of oxaliplatin (1.5 mg/kg body weight/3 times per week), or a full dose of oxaliplatin (3 mg/kg body weight/3 times per week). The control (sham) group received a standard chow diet. Treatments started three weeks before and continued for three weeks after orthotopic CRC induction. The number of metastases, tumour weight and volume were quantified ex-vivo. The expression of cytochrome c, cleaved caspase-9 and -3, DNA damage, PD-L1, PD-L2 and HSP-70 were determined. Results: A significant reductions in the weight and volume of tumours were observed in mice treated with KO and KO plus a ½ dose of oxaliplatin compared to the sham group, similar to oxaliplatin-treated mice. KO, and KO plus ½ dose of oxaliplatin significantly increased the expression of cytochrome c, cleaved caspase-9 and -3, and DNA damage and decreased expression of PD-L1, PD-L2 and HSP-70 in tumour tissues compared to the sham group. Conclusions: The in vivo anti-cancer effects of KO are comparable with oxaliplatin. Thus, dietary KO supplementation has a great potential as a therapeutic/adjunctive agent for CRC treatment

Ehogenicity of thyroid gland on ultrasonography in primary hypothyroidism

Background. The aim of this study was to investigate the association between echogenicity of thyroid gland and low thyroid function. Methods. The study group included 60 patients, with normal and low thyroid function, who visited the Outpatient Department of the Clinic of Endocrinology, Diabetes and Metabolic Disorders in the period from April 2008 to October 2009. Echogenicity of the thyroid gland in all patients was evaluated by ultrasonography as well as serum concentrations of TSH, fT4 and thyroid autoantibody (anti-TPO). To find out the association between thyroid ehogenicity with mean TSH and anti-TPO in different ages, we divided the patients into five subgroups according to age. Results. Patients with decreased echogenicity had a higher mean TSH compared with patients with normal echogenicity (2.77 mIU/l vs. 1.75 mIU/l) (p=0.04). Differences were more significant in patients with markedly decreased echogenicity (6.34 mIU/l vs. 1.75 mIU/l) (p< 0.0001). Patients with reduced echogenicity had a higher risk of having anti-TPO than patients without normal echogenicity (p<0.001). This association was stronger when echogenicity was markedly decreased. According to age, only younger population (19-29 years) with decreeased and markedly decreased echogenicity had significantly higher mean TSH and anti-TPO values. Conclusions. Thyroid ultrasonography changes can be used as an early sign of low thyroid function, especially in younger population

Maternal lipids may predict fetal growth in type 2 diabetes mellitus and gestational diabetes mellitus pregnancies

Aim: During diabetic pregnancy, complex metabolic changes occur in the lipid profile. The aim of the study was to determine the predictive values of maternal serum lipid levels on large-for-gestational age newborns during the third trimester in pregnancies of women with type 2 diabetes mellitus (DM2) and gestational diabetes mellitus (GDM). Material and methods: Data of forty-three pregnancies of women with DM2 and two hundred women with GDM were analyzed. The analysis encompassed the following parameters: age, body mass index (BMI), lipid parameters, HbA1c in first, second and third trimester of pregnancy, preeclampsia and baby birth weight. Results: DM2 and GDM groups showed statistically significant differences in the following variables: total lipids, triglycerides, total cholesterol, BMI, age, baby birth weight, incidence of SGA and preterm delivery (9.4 ± 2.3 vs. 11.0 ± 2.3 mmol/L, 2.4 ± 1.4 vs. 3.4 ± 1.6 mmol/L, 5.5 ± 1.2 vs. 6.4 ± 1.4 mmol/L, 30.6 ± 5.4 vs. 26.9 ± 5.2 kg/m2, 34 ± 7.8 vs. 31.5 ± 5.6 years, 3183 ± 972 vs. 3533 ± 699 g., 20% vs. 7.5%, 27.9 vs. 14%, respectively, p < 0.05). Linear multiple regression analysis demonstrated that triglycerides, LDL-C and total cholesterol were independent predictors of LGA (p < 0.05). Conclusion: Triglycerides and LDL-C in the third trimester of pregnancy are independent predictors for fetal macrosomia in DM2 and GDM pregnancies. Thus, the maternal serum triglycerides and LDL-C levels determined in the maternal blood taken in the third trimester of pregnancy may indentify women who will give birth to LGA newborns

Glycosylated Haemoglobin and Adverse Pregnancy Outcomes in Women with Diabetes Mellitus Type 1 Compared to Pregnancies in the General Population

Aim: to compare pregnancy outcomes in type 1 diabetic pregnancies with the general population and the impact of glycemic control (HbA1c) on pregnancy outcomes. Material and methods: The study group included 55 consecutive pregnant women with type 1 diabetes who attended the Outpatient department of Endocrinology, Diabetes and Metabolic disorders in their first trimester of pregnancy. For comparison, data of 22767 deliveries in 2004 year were used, published in “Perinatal outcomes in Republic of Macedonia for year 2004”. The following parameters were studied: age, maternal body mass index, diabetes duration, planning of pregnancy, HbA1c values in first, second and third trimester, pre-eclampsia, birth weight, gestational age at delivery, mode of delivery, and pregnancy outcomes (spontaneous abortions, perinatal mortality, and major congenital malformations). Results: The perinatal mortality rate was 10,9% and congenital malformation rate was 7,3%, in type 1 diabetic pregnancies compared with 1,1% and 1,9%, respectively in the general population. Macrocosmic babies were born in 9,1% of type 1 diabetic pregnancies compared with 1,01% in the general population. The caesarean section rate was 61,8% and 16,1%, respectively, and the preterm delivery rate was 25,5% and 6,1%, respectively. Pregnancies with serious adverse outcomes (spontaneous abortion, perinatal mortality and congenital malformations) were characterized by higher Values of HbA1c in the first trimester of diabetes type 1 pregnancies Conclusion: Type 1 diabetic pregnancies have worse outcomes then general population. Planning of pregnancy and rigorous glycaemic control using glucose monitoring before and during early pregnancy are crucial for better pregnancy outcomes. Key words: pregnancy outcomes, diabetes mellitus type 1, glycated haemoglobin, congenital malformations