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Antimicrobial resistance (AMR) is an increasing global concern in small animal veterinary practices. The overuse and misuse of antimicrobials in companion animals can contribute to the spread of AMR between animals and humans. This study aimed to assess small animal veterinarians’ knowledge, attitudes, and behaviors towards antimicrobial use (AMU) and AMR in North Macedonia. A web-based questionnaire was developed, including questions addressing demographics, owner influence on antibiotic prescription and prescribing practices. The response rate of the small animal veterinarians was 47.50% (57/120), with most of them (84.20%) having 1-15 years of work experience. Pearson’s chi-square test of independence was used for contingency tables that met the chi-square assumption, and Fisher’s exact test was conducted for contingency tables that did not. Most veterinarians responded that they had perceived pressure and direct demands from pet owners to prescribe antimicrobials (35.09% frequently, 45.61% occasionally). In the survey, 50.77% of the veterinarians reported frequently administering antibiotics without conducting bacteriological or antimicrobial susceptibility testing. They typically resorted to these tests in cases with recurrent infections, such as skin infections (19.63%), or when empirical therapy is ineffective (14.72%). Based on the findings, it can be concluded that veterinarians play a crucial role in tackling the AMR in North Macedonia. The lack of stewardship programs or guidelines for responsible antimicrobial use in companion animals is a critical issue that requires urgent attention, emphasizing the vital role of veterinarians in combating AMR.

Introduction: Tumor budding (TB) is considered to be a morphological and prognostic factor relevant to colon cancer (CC). The aim of our study is to assess the TB and to evaluate its relationship to clinicopathological findings within stage II and III CC patients as a single center experience.

The present study aimed to determinate the effect of external progesterone (P4) supplementation on luteolysis in cows under heat stress. Forty-eight (n=48) dairy cows in the period from July–September 2018 were part of and at day 35±3 postpartum scored for BSC, synchronized using PG-3-G + Ovsynch protocol and randomly allocated into two treatments: PRID group (n=27) treated with external P4 device between G1 and PGF2αand CON group (n=21) left without treatment. Collection of blood samples to assess P4 concentrations was done at Pre-PG, at G1, at PGF2α, at 72 h after PGF2α(at timed artificial insemination TAI) and at d 21 after TAI. The pregnancy diagnosis was done at d 21 and d 30 after TAI by ultrasound. The average temperature-humidity index (THI) was 79.5±0.6. At G1, the P4 was significantly lower in the PRID group (1.84±0.99 ng/mL) in comparison to the CON group (2.97±1.82 ng/mL). In contrast, at PGF2α, there was a tendency (p=0.09) of increased P4 concentration in PRID group compared with the CON group (4.26±1.68 and 3.74±2.39 ng/mL), respectively. At TAI, more PRID cows (p=0.0001) had a lower P4 (0.06±0.03 ng/mL), in comparison to CON (1.28±2.41 ng/mL). At d 21 and d 30 after TAI, more PRID cows were predicted and diagnosed pregnant (16/27 or 59.25% and 13/27 or 48.14%) compared with the CON group (11/21 or 52.38% and 8/21 or 38.08%) respectively, but without any significant differences. Supplementation of the P4 during the Ovsynch protocol increases the P4 before TAI and reduces the incomplete luteolysis in heat stressed dairy cows.

Infectious hematopoietic necrosis (IHN) is a common disease in the intensive production of salmonids. The IHN virus (IHNV) was isolated for the first time in North Macedonia in 2018. This study aimed to investigate the distribution and genotype of IHN in farmed rainbow trout and autochthonous salmonid fish in North Macedonia following the first detection. The samples were collected from 47 trout farms. Trout fry with or without clinical signs of IHN were selected as individual samples. Kidney, spleen, and heart were taken from each fish during the dissection. Three pooled samples were collected from each farm. A total of 141 pooled samples were collected: Rainbow trout (Oncorhynchus mykiss) n=127, Macedonian trout (Salmo macedonicus) n=11, and Ohrid trout (Salmo letnica) n=3. The virus was detected in 43 samples (30.50%): rainbow trout (n=40), Macedonian trout (n=2), and Ohrid trout (n=1). There were 18 (38.30%) positive fish farms. The MAKIHNV1 isolate from 2018 (MN641902) and the newly isolated virus shared a similarity of >99 and were placed in clade E-1 of European genogroup E. The IHN has spread throughout the country and is also present in the autochthonous salmonids.

Hyperadrenocorticism (HAC) in dogs is routinely treated with trilostane single-dose (CTG) which is reported to cause adverse reactions. The current retrospective study of several dogs with terminal stage of HAC aimed to compare the clinical, hematological, and biochemical effects of trilostane titration-dose treatment (TTG) with the single-dose treatment (CTG). All clinical cases (n=7) were confirmed on HAC by anamnestic, clinical, hematology, biochemistry, and low-dose dexamethasone suppression test findings, indicative for Cushing’s disease. Two cases were treated with CTG (2.2-6.7 mg/kg, single dose daily) and their treatment was discontinued on the second week due to adverse reactions. The TTG cases were treated for up to 12 weeks (0.5 mg/kg once daily for 7 days, and then with 0.5 mg/kg twice daily for 7 days). Blood samples and clinical checks were performed on 0., 4., and 12. weeks of the treatment. Hemoglobin was non-significantly higher in TTG at 12 weeks. Alanine transaminase was significantly lower in the TTG cases on the 12. week of the treatment (78.04±15.37 U/L) compared to the 0-week (137.81±24.03 U/L), and 4-week samples (131.92±23.36 U/L). No significant differences were observed with the CTG cases. Alkaline phosphatase was significantly lower on 12-week samples in TTG (251.02±93.06) compared to the 4-week (567.94±283.93 U/L), and 0-week samples (1,341.84 U/L). In conclusion, TTG has indicated to have significantly higher tendency to decrease alanine transaminase and alkaline phosphatase, alleviating the negative effects on the liver. The clinical findings were more adverse for the CTG.

Emphysematous cholecystitis is reported to have a low incidence of less than 1% in all cases of acute cholecystitis and yet a high mortality rate of up to 15%. It is most commonly seen in male diabetic patients with advanced age. The diagnosis is established with the presence of gas in the gallbladder lumen and/or within its wall which can be seen on plain abdominal radiography, abdominal ultrasound, and abdominal computerized tomography. The clinical presentation refers to one of acute cholecystitis, but the treatment requires prompt cholecystectomy since the patient’s condition can deteriorate due to the possibility of gallbladder perforation. We present a case of a 71-year-old female diabetic patient with calculous emphysematous cholecystitis treated with emergency open cholecystectomy.

Aim: Lymphovascular Invasion (LVI) and Perineural Invasion (PNI) represent undesirable but still realistic pathological features of rectal cancer, associated with poor prognosis and worse survival. The aim of this study is to assess the incidence of LVI and PNI in patients treated for rectal cancer and the impact of LVI and PNI on patient survival.

Brilliant cresyl blue (BCB) staining is a method used for selection of developmentally competent porcine oocytes, in addition to the standard morphological classification. The aim of the current study was to assess the relationship between BCB staining in porcine oocytes with high and low morphological characteristics and its maturation rate. In the first part of the study, cumulus-oocytes complexes (COCs) (n=434) were aspirated from 60 ovaries. According to the morphological classification and BCB staining, they were divided in 4 groups: COC+/BCB+, COC−/BCB+, COC+/BCB−, and COC−/BCB−. In the second experiment, COCs (n=445) were categorized into 3 groups: control, BCB+, and BCB−. Significant differences in the maturation rate were observed between COC+/BCB+ and COC−/BCB− (66% vs. 23%), and between COC+/BCB+ and COC−/BCB+ (66% vs. 38%) (p<0.0001). Significant differences were also registered between COC−/BCB+ and COC−/BCB-(38% vs. 23%) and between COC+/BCB− and COC−/BCB− (53% vs. 23%) (p<0.01). Results from the second experiment showed that only BCB− oocytes had a significantly lower maturation rate (28%) compared to the control (63%) and BCB+ oocytes (59%) (p<0.001). These findings showed that COCs with high morphological characteristics had higher developmental ability compared to COCs with low morphology. The BCB-staining in high-quality oocytes did not have a significant impact on their maturation ability compared to a control group, but it might be useful for selecting developmentally competent oocytes with low morphology characteristics.

The purpose of this in-vitro study was to investigate the fracture resistance and failure mode of porcelain veneers using three preparation designs.

Chronic solitary ulcers over the tongue are challenging to diagnose as they are caused by various etiological factors. Differential diagnoses are chronic traumatic ulcers, ulcers due to squamous cell carcinoma, syphilis, fungal, viral, and tuberculosis (TB).

Electronic components may contain sensitive data, either on the storage chip or on the microprocessor. Deliberate and precise perturbation of their operation or measurement of their activity through auxiliary channels can be used to extract the secrets they contain. For these reasons, it is important that the integrity of the integrated circuit is ensured. It is therefore necessary to implement protection mechanisms against threats such as physical attacks. In this paper, we present a new device for protection against and supervision of fault injection and electromagnetic listening attacks. It allows the detection of several types of physical attacks and provides an appropriate response for each type of threat.

The incidence of clinical mastitis (CM) in small and large-scale dairy cow farms can be highly variable and can be affected by age, parity, post-calving status, and atmospheric conditions. The current study aimed to assess the CM-incidence and its association with the post-calving days, number of lactations, season, and number of affected udder quarters in dairy cows in small-scale dairy farms. The study was conducted within one calendar year in 177 small-scale farms with 864 dairy cows. Clinically confirmed CM cows (n=72) were sampled from each udder quarter and processed for bacteriology examination. The CM-positive samples were grouped according to the season (spring, summer, autumn, and winter), the number of days after calving (<90, 90-180, and >180), the number of lactations (1-st, 2-5-th, and >6-th), and the number of affected udder quarters (one, two, three, and four). The CM-positive samples (n=72, 8.3%) were confirmed on bacteriology examination in significantly lower count (n=56, 6.5%). The 2-5-th lactations cows (n=35, 68.6%) were significantly more compared to the first lactation (n=6, 11.8%), and >6-th lactation cows (n=10, 19.6%). CM cows with infection of one udder quarter (n=40, 78.4%) were significantly more than the cows with two (n=6, 11.8%), and four infected quarters (n=5, 9.8%). The CM-incidence in small-scale dairy cow farms in North Macedonia was 8.3% and 6.5% by clinical and bacteriology examination, respectively. The highest CM susceptibility was observed in the cows between the second and fifth lactations. One udder quarter was most frequently affected in CM-positive cases.

AbstractIntroduction:Poor outcome is still a challenging concern in patients with out-of-hospital cardiac arrest (OHCA) world-wide and there are large differences between European countries regarding not only incidence rates, but survival rates as well. In 2014, Serbian Resuscitation Council initiated regular data collection on epidemiology of OHCA, according to the European Registry of Cardiac Arrest (EuReCa) study protocol.Study Objective:The aim of this study is to analyze the results of the first five-year period after initiation of EuReCa study protocol elements implementation in OHCA epidemiological data collection in Serbia.Methods:The observed period in this study is about the data on OHCA, collected within the observed area of 16 municipalities covering 1,604,015 citizens, during the period from October 1, 2014 – December 31, 2019. The study included data on all-cause OHCA in both adult and pediatric patients, according to the EuReCa One study protocol, of which all segments were observed.Results:Within the study period, 5,196 OHCA patients were observed with annual incidence of 83.60/100,000. Of all registered events, 43.9% were witnessed. The most common collapse location was patient’s residence (88.7%). Within the group of initiated cardiopulmonary resuscitation (CPR), cardiac etiology was observed in 80.5% of cases and shockable rhythm in 21.7%. Return of spontaneous circulation (ROSC) prior to hospital admission was significantly more frequently achieved and maintained on admission in witnessed cases, cases occurring out of patient’s residence, and in cases with shockable initial rhythm (P <.01).Conclusion:The OHCA incidence in Serbia is comparable with the incidence in the majority of European countries, and survival rates are now significantly higher in Utstein events compared to previous results from Serbia. Enrolment of witnessing bystanders in initiating CPR measures remains a concern requiring effort towards understanding of CPR initiation importance and education of general population in administering CPR measures.

Intraoral scanners have become increasingly popular due to their convenience, accuracy and time-saving capabilities. The aim of this study was to compare the clinical outcomes of zirconia crowns made with two different intraoral scanners.

Dental caries is considered to be the result of the disturbance of the ecological balance of the oral cavity, rather than the result of poor nutrition or insufficient oral hygiene. The aim of our research is to determine the relationship between some salivary parameters and dental caries in a group of respondents with primary and permanent teeth.

Alveolar ridge preservation can be achieved with three types of grafts: soft tissue, hard tissue, or a combination of soft and hard tissue. Nowadays, a technique so called socket shield (SST) which includes a part of the dental root left into the socket, is one of the most popular methods for its esthetic outcome especially in dental implantology.

High-grade serous ovarian cancer (HGSOC) is an archetypal cancer of genomic instability1–4patterned by distinct mutational processes5,6, tumour heterogeneity7–9and intraperitoneal spread7,8,10. Immunotherapies have had limited efficacy in HGSOC11–13, highlighting an unmet need to assess how mutational processes and the anatomical sites of tumour foci determine the immunological states of the tumour microenvironment. Here we carried out an integrative analysis of whole-genome sequencing, single-cell RNA sequencing, digital histopathology and multiplexed immunofluorescence of 160 tumour sites from 42 treatment-naive patients with HGSOC. Homologous recombination-deficient HRD-Dup (BRCA1mutant-like) and HRD-Del (BRCA2mutant-like) tumours harboured inflammatory signalling and ongoing immunoediting, reflected in loss of HLA diversity and tumour infiltration with highly differentiated dysfunctional CD8+T cells. By contrast, foldback-inversion-bearing tumours exhibited elevated immunosuppressive TGFβ signalling and immune exclusion, with predominantly naive/stem-like and memory T cells. Phenotypic state associations were specific to anatomical sites, highlighting compositional, topological and functional differences between adnexal tumours and distal peritoneal foci. Our findings implicate anatomical sites and mutational processes as determinants of evolutionary phenotypic divergence and immune resistance mechanisms in HGSOC. Our study provides a multi-omic cellular phenotype data substrate from which to develop and interpret future personalized immunotherapeutic approaches and early detection research.

Aim: To evaluate the effect of extralevator abdominoperineal excision of the rectum (ELAPE) on the circumferential resection margin (CRM) and overall survival in comparison to standard abdominoperineal excision of the rectum (APE) in patients with advanced rectal cancer.

Artificial insemination (AI) as a part of assisted reproductive technologies represents the oldest and most widespread method used to accelerate genetic progress in all domestic animals. After its first implementation in ovine reproduction and almost 80 years afterward, AI is continuously used for improving the genetic merit, utilizing either fresh or short-time chilled semen. Nevertheless, regardless of the semen used for insemination, the conception rate (CR) is still lower in comparison to natural service. At least two factors are commonly thought to limit the success of the AI and reduce the CR: (1) failure of placing the semen directly into the uterus due to the specific anatomic structure of the ewe’s cervix; (2) lower viability of ram spermatozoa during cryopreservation (<30% progressively motile spermatozoa after thawing). This review elaborates on recent studies that aimed to achieve acceptable CR through the implementation of cervical or intrauterine insemination: deep intracervical, intrauterine trans-cervical, and intracornual. Several hormonal treatments (oxytocin, estrogen, or prostaglandin) were evaluated on inducing cervical dilation that facilitates insemination. A comprehensive analysis was given to the effects of several antioxidants (GSSG, GSH, and cysteine) supplemented in ram semen-freezing media. Sex-sorted ram semen fertility rate results were presented from our studies.

CT imaging and disease-specific structured reporting are essential to guide primary treatment selection in patients with advanced ovarian cancer.

Aim: To determine and define the advantages of the laparoscopic appendectomy in the treatment of complicated appendicitis by comparing it with the open appendectomy.

Cystic echinococcosis is a cosmopolitan zoonotic infection, commonly seen in areas where livestock and dogs are kept together. The objective of this study was to investigate the prevalence of echinococcosis in slaughtered cattle and sheep in North Macedonia and to evaluate the location and fertility/sterility rates of the cysts. A total of 302 slaughtered animals were examined with visual inspection, palpation, and incision of the internal organs in order to detect cysts. Microscopic examination was carried out to determine the presence of protoscoleces. The overall prevalence of echinococcosis in slaughtered cattle and sheep was 60%. The presence of cysts increased with age in cattle. In both cattle and sheep, the most common affected organs were the liver and lungs infected with more than one cyst. The findings showed higher fertility rate in sheep which confirmed their role as the most important intermediate host. This study shows that North Macedonia is an endemic region, and serious control strategies should be implemented, with special emphasis on safety disposal of infected organs and anthelmintic treatment of the dogs.

The local extent of prostate cancer on MRI is independently associated with clinically relevant long-term oncologic outcomes after radical prostatectomy.

Hepatocellular carcinomas with nuclear factor E2–related factor 2 (NRF2) mutation had rapid progression after transarterial embolization and showed resistance to ischemia, but also became more sensitive to NRF2 inhibition in the setting of ischemia.

Many techniques used and still in usage for solving the problem of islanding detection are intrinsically passive, active, or hybrid of both. Each one of them has its own benefits and drawbacks. In this paper, we propose a method, which takes the advantage of a machine learning (ML)-based algorithm, namely, support vector machine (SVM), in order to produce the results more efficiently. The results of the simulations based on the model and experimentally measured parameters of a real-life practical photovoltaic (PV) plant give much better output than the traditional reported methods. During the tests and simulations, an additional problem, namely, grid fault, emerged, posing new challenges for the proposed method. Occurrences of islanding and grid fault are grouped together with the same kernel dimension and no custom hyperplane bordering. Discrimination between islanding and grid fault events is an essential dilemma, which is handled by the proposed SVM-based algorithm to achieve more precision in islanding detection and simultaneously detect the grid faults authentically. Nondetection zones (NDZs) and detection time (DT) are tested using two dimensions, namely, the generated active energy from PV plant (0%–110% of $P_{n}$ ) and distribution network voltage levels (±10% of $U_{n}$ ). Simulations based on the model and parameters of a real-life practical PV power plant are performed in MATLAB/Simulink environment, and several tests are executed for several scenarios. Finally, comparisons with previously reported techniques prove the effectiveness, authenticity, selectivity, accuracy, and precision of the proposed islanding and grid fault detection strategy with allowable impact on power quality according to UL1741 and its superiority over other methods.

In metastatic cancer, the degree of heterogeneity of the tumor microenvironment (TME) and its molecular underpinnings remain largely unstudied. To characterize the tumor–immune interface at baseline and during neoadjuvant chemotherapy (NACT) in high-grade serous ovarian cancer (HGSOC), we performed immunogenomic analysis of treatment-naive and paired samples from before and after treatment with chemotherapy. In treatment-naive HGSOC, we found that immune-cell-excluded and inflammatory microenvironments coexist within the same individuals and within the same tumor sites, indicating ubiquitous variability in immune cell infiltration. Analysis of TME cell composition, DNA copy number, mutations and gene expression showed that immune cell exclusion was associated with amplification of Myc target genes and increased expression of canonical Wnt signaling in treatment-naive HGSOC. Following NACT, increased natural killer (NK) cell infiltration and oligoclonal expansion of T cells were detected. We demonstrate that the tumor–immune microenvironment of advanced HGSOC is intrinsically heterogeneous and that chemotherapy induces local immune activation, suggesting that chemotherapy can potentiate the immunogenicity of immune-excluded HGSOC tumors.

The present study aimed to evaluate whether the induction and the formation of an accessory corpus luteum (CL) after AI might increase the pregnancy per AI (P/AI) in heat stressed dairy cows. Starting at d 50±3 post-partum, 113 lactating Holstein cows from one commercial herd during summer were scored for body condition, blood sampled and examined by ultrasound. Those bearing a CL>25mm and progesterone (P4) level>2ng/mL were synchronized using a double PGF2α injection given 12 h apart and AI-ed at detected estrus. In total 18 cows, there were not any signs of estrus (n=10) nor a P4 level <2ng/mL at the time of enrolment (n=8) and therefore they were excluded from the study, leading to 95 cows finally enrolled.. At d5 post-AI, cows were randomly allocated into 2 groups: control group (CON, n=45) without any additional treatment, and treatment group (GnRH, n=50), treated with 0.008 mg Buserelin – a GnRH agonist. Blood sampling and ultrasound examination were done at d5, d14 and at d21 after AI, whereas the pregnancy diagnosis was done at d21 and d30 after AI. Average daily temperature and relative humidity values were used to calculate the temperature-humidity index (THI). The average THI during the experiment was 79.5±0.6. At d5, no differences were observed neither between the number of the CL nor between the P4 level in both groups. At d14 and d21, 82% of the GnRH-treated cows had more than one CL versus 0% of the CON cows. Both at d14 and 21, GnRH-treated cows had higher P4 levels compared to the CON cows (p<0.05). In addition, P/AI were higher in the GnRH group than in the CON group (65% vs. 48.3%, p<0.05), whereas late embryonic losses were higher in CON in comparison to GnRH cows (10.6 vs. 4%, respectively). The BCS at the moment of insemination did not affect P/AI (p>0.05). In conclusion, the induction of an accessory CL at d 5 after AI might increase P/AI in heat stressed dairy cows.

The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3gene. The two most common gene variants in the HF breed are CSN3*Aand CSN3*Bwhile CSN3*Ehas been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A(0.584), CSN3*B(0.336) and CSN3*E(0.08). The observed genotype frequencies differed significantly (P<0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*Ballele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.

This paper presents a new approach for determining the impact of lightning strike currents on transmission network elements failures, based on fuzzy logic (FL) and expert systems. The location of lightning strike is determined by means of lightning location system (LLS) and failures in the transmission network, sorted by type of the equipment, are obtained from supervisory control and data acquisition (SCADA) system. The input data set includes two sets. The first set consists of lightning strike locations and current values between the cloud and the ground. The second set consists of current values from SCADA system before and after the fault, protection tripping information, and the state and position of the switches. The proposed FL-based solution is based on a fuzzy decision-making system (DMS), including both data sets in order to provide a power system operator (PSO) with a precise and accurate decision needed in time of emergency. The described model has been tested for functionality and correct results have been obtained, which confirms the membership function (MF) assessment and proves the efficiency and authenticity of the proposed DMS.

The question of the islanding operation of distributed generations (DGs) and that of the current inadmissible networking policy of islanding operations sets a demanding task for an efficient islanding detection, preferably from a single place, using either passive or active islanding-detection method. Islanding detection shall be faster than the fast automatic reclosing of the connected transmission line connected to the DG unit. This paper deals with the detection method of islanded operation mode for the large power synchronous generator-based DGs in bio-mass power plants. The proposed method has been applied to a practical 10-MW bio-mass power plant and experimentally tested to verify the simulation results. The rate of change of reactive power (dQ/dt >) criterion is derived as a yardstick for islanding detection. The practical power plant model has been implemented to be simulated in DIgSILENT PowerFactory and MATLAB/Simulink software environments. Offline digital time-domain simulations and experimental tests on a practical bio-mass power plant reveals that the proposed method efficiently, and authentically detects islanding phenomena with high accuracy, low detection time, very small number of non-detected zones and, with almost zero power balance (ΔP = 0).

Thermal ablation of small renal masses is increasingly accepted as an alternative to partial nephrectomy, and an understanding of preablation planning, postablation imaging findings, and potential complications is essential for successful ablation.

Reduced glutathione (GSH) and homologous ram seminal plasma (HSP), used as additives in cryopreserving (CP) media prior to freezing, showed conflicting results in retaining structural integrity and progressive motility in post-thawed ram spermatozoa. The aims of this research were: (1) to assess the effect of GSH and/or HSP supplementation via soybean-lecithin CP extender on cryopreserved ram spermatozoa viability, morphology and motility pattern; and (2) to assess the effect of incubation in the context of the previous aim. Quantitatively and qualitatively, homogenized and pooled ram ejaculates (N=10) were extended with one of the following extenders: control (C) – tris-based, GSH and HSP-free, experimental-1 (E1) – C + GSH 5 mM, experimental-2 (E2) – C + HSP 20 % and experimental-3 (E3) - GSH 5 mM + HSP 20 %. Following thawing, samples were taken at 0- and 3-hours from each group (n=10) and were assessed for spermatozoa viability, morphology, and motility pattern. C-0h samples yielded a spermatozoa population with low viability, altered head morphology and highly deviated motility pattern. E3-3h samples yielded spermatozoa with unaffected viability, head morphology and high progressive motility. In conclusion, E3 extender added to cryopreserved-thawed ram spermatozoa is most efficient in obtaining high viability, unaltered head morphology, and progressive motility.

Masses in the female pelvis can present a diagnostic challenge, but a systematic anatomy-based approach to MR image interpretation can help one navigate the spaces of the female pelvis and commonly encountered lesions in those spaces with greater confidence.

We have constructed and structurally characterized a Pseudomonas aeruginosaazurin mutant Re126WWCuI, where two adjacent tryptophan residues (W124 and W122, indole separation 3.6–4.1 Å) are inserted between the CuIcenter and a Re photosensitizer coordinated to the imidazole of H126 (ReI(H126)(CO)3(4,7-dimethyl-1,10-phenanthroline)+). CuIoxidation by the photoexcited Re label (*Re) 22.9 Å away proceeds with a ∼70 ns time constant, similar to that of a single-tryptophan mutant (∼40 ns) with a 19.4 Å Re–Cu distance. Time-resolved spectroscopy (luminescence, visible and IR absorption) revealed two rapid reversible electron transfer steps, W124 → *Re (400–475 ps, K1≅ 3.5–4) and W122 → W124•+(7–9 ns, K2≅ 0.55–0.75), followed by a rate-determining (70–90 ns) CuIoxidation by W122•+ca. 11 Å away. The photocycle is completed by 120 μs recombination. No photochemical CuIoxidation was observed in Re126FWCuI, whereas in Re126WFCuI, the photocycle is restricted to the ReH126W124 unit and CuIremains isolated. QM/MM/MD simulations of Re126WWCuIindicate that indole solvation changes through the hopping process and W124 → *Re electron transfer is accompanied by water fluctuations that tighten W124 solvation. Our finding that multistep tunneling (hopping) confers a ∼9000-fold advantage over single-step tunneling in the double-tryptophan protein supports the proposal that hole-hopping through tryptophan/tyrosine chains protects enzymes from oxidative damage.