The impact of neoadjuvant systemic treatment on postoperative complications in breast cancer surgery.

PURPOSE: The aim of the study was to analyze the impact of neoadjuvant systemic treatment (NST) on postoperative complications and the beginning of adjuvant treatment. METHODS: This study includes data from a prospectively maintained database including patients with breast cancer (BC) stage I-IV with or without NST undergoing breast cancer surgery between January 2010 and September 2021. RESULTS: Out of 517 enrolled patients, 77 received NST, 440 had primary breast surgery. After NST patients underwent surgery after a meantime of 34 days (26.5-40 days). No statistical significance could be found comparing the complication grading according to the Clavien Dindo classification. The complications were most frequently rated as grade 3b. There were no complications with grade 4 or higher. When differentiating into short and long-term, the overall rate of short-term complications was 20.3% with no significant difference between the two groups (20.8% vs. 20.2%). Regarding long-term complications, there was more impairment of shoulder mobility (26.0% vs. 9.5%, p ≤ 0.001) and chronic pain (42.9% vs. 28.6%, p ≤ 0.016) for patients with NST. The beginning of the administration of the adjuvant treatment was comparable in both groups (46.3 days vs. 50.5 days). CONCLUSION: In our cohort, complications between both groups were comparable according to Clavien Dindo. This study shows that NST has no negative impact on postoperative short-term complications and most importantly did not lead to a delay of the beginning of adjuvant treatment. Therefore, NST can be safely admitted, even when followed by extensive breast reconstruction surgery.

Adhesion of Malassezia pachydermatis of different growth type to canine epithelial cells.

A total of 100 Malassezia pachydermatis strains recovered from skin and mucosal membranes of dogs were evaluated for their adhesive properties. Two types of growth, related to colony morphology on Sabouraud agar, were observed (type I and II). The mean number of fungal cells attaching to canine buccal epithelial cells was found to be 17. The number of adhered cells was greater (statistically significant at the level of p < 0.01) in strains belonging to the type I.

Effect of age and stage of lactation on whey protein content in milk of cows of different breeds.

The aim of the study was to determine the effect of age of cows, i.e. subsequent lactation and stage of lactation, on bioactive whey protein content in milk of dairy cows of six primary breeds kept in Poland. In all cases the significant correlations between lactoferrin concentration in milk and stage of lactation was stated. Its content gradually increased with the course of lactation but the changes in the content of this protein were highly dependent on breed of cows, what was indicated also by high interactions between breed and stage of lactation.

Role of IncP-1ß plasmids pWDL7::rfp and pNB8c in chloroaniline catabolism as determined by genomic and functional analyses.

Broad-host-range catabolic plasmids play an important role in bacterial degradation of man-made compounds. To gain insight into the role of these plasmids in chloroaniline degradation, we determined the first complete nucleotide sequences of an IncP-1 chloroaniline degradation plasmid, pWDL7::rfp and its close relative pNB8c, as well as the expression pattern, function, and bioaugmentation potential of the putative 3-chloroaniline (3-CA) oxidation genes. Based on phylogenetic analysis of backbone proteins, both plasmids are members of a distinct clade within the IncP-1ß subgroup. The plasmids are almost identical, but whereas pWDL7::rfp carries a duplicate inverted catabolic transposon, Tn6063, containing a putative 3-CA oxidation gene cluster, dcaQTA1A2BR, pNB8c contains only a single copy of the transposon. No genes for an aromatic ring cleavage pathway were detected on either plasmid, suggesting that only the upper 3-CA degradation pathway was present. The dcaA1A2B gene products expressed from a high-copy-number vector were shown to convert 3-CA to 4-chlorocatechol in Escherichia coli. Slight differences in the dca promoter region between the plasmids and lack of induction of transcription of the pNB8c dca genes by 3-CA may explain previous findings that pNB8C does not confer 3-CA transformation. Bioaugmentation of activated sludge with pWDL7::rfp accelerated removal of 3-CA, but only in the presence of an additional carbon source. Successful bioaugmentation requires complementation of the upper pathway genes with chlorocatechol cleavage genes in indigenous bacteria. The genome sequences of these plasmids thus help explain the molecular basis of their catabolic activities.

Changes of protein content and its fractions in bovine milk from different breeds subject to somatic cell count.

Global milk production is undeniably dominated by 2 dairy breeds recognized worldwide: Holstein-Friesian and Jersey. A third breed, Simmental, serves as a dual-purpose breed. The objective of the present research was to establish potential changes in the fractional components of bovine milk protein (mainly whey) in relation to the health status of a dairy cow's mammary glands, which is closely determined by somatic cell count (SCC). The milk of 3 breeds was studied: Polish Holstein-Friesian (Black and Red-White varieties), Simmental, and Jersey. The cows were housed in freestall barns and fed according to the total mixed ration feeding system for both winter and summer periods. Milk samples were collected individually from each cow twice a year, in the winter and summer seasons. A total of 1,822 milk samples were evaluated (946 in winter and 876 in summer). The milk was examined for SCC, crude protein, casein, and whey fraction proteins (α-lactalbumin, ß-lactoglobulin, lactoferrin, BSA, and lysozyme). The research material for each breed was split into 4 groups based on SCC (group I: ≤100,000 cells/mL; group II: 101,000 to 400,000 cells/mL; group III: 401,000 to 500,000 cells/mL; and group IV: 501,000 to 1,000,000 cells/mL). It was found that an increase of SCC promulgated a progressive decline in the daily yield of milk, which was significantly true for the Polish Holstein-Friesian. The level of crude protein decreased slightly as SCC increased, and casein concentration (r=-0.591) also followed this trend of decline. Elevation of SCC produced a decrease of major albumins (i.e., α-lactalbumin and ß-lactoglobulin). However, SCC increase induced an increase in the immunoactive proteins (lactoferrin and lysozyme) as well as BSA. The interactions of a breed with increased SCC, which can be measured based on the BSA content of the milk, has indicated various levels of susceptibility to the increase in different breeds. This is confirmed by different values of correlation coefficients for these relationships: 0.71 in the Holstein-Friesian, 0.58 in Simmental, and 0.47 in the Jersey cows. Holstein-Friesian cows are more sensitive to mammary gland infections causing a greater decline of their daily milk yields, which, in turn, is reflected in an increase of the negative value of the correlation coefficients between SCC and milk efficiency (-0.24). In the other 2 breeds, the correlations were also negative, but substantially lower (-0.12 and -0.15).

Constant darkness negatively affects the outcome of hormonally induced reproduction in cultured Eurasian perch females.

This study aimed to assess the effect of constant darkness applied to fish during controlled breeding on reproductive traits in domesticated females of Eurasian perch. Based on the assumption that keeping fish in constant darkness during the reproduction operation may reduce stress, suspected to be responsible for variable spawning effectiveness in this species. Two conditions were assessed (16 h light per day [group 16L] and constant darkness [group 0L], two tank replicates per condition). The reproductive protocol involved a 7-day-long adaptation period for group 0L where photoperiod was reduced by 2.3 h a day down to constant darkness. After the adaptation period, two hormone injections (salmon gonadoliberin analogue) were applied to both groups: priming (10 µg/kg) and resolving (25 µg/kg) with a 7-day interval between them. During the study, morphometric indices were recorded and blood, brain, and pituitary samples were collected to assess stress markers and determine hypothalamic-pituitary-gonadal axis functioning via measuring blood plasma hormones, as well as gonadoliberin and gonadotropins (luteinising hormone [LH] and follicle-stimulating hormone [FSH]) transcript abundance (n = 7 for each group at each sampling point). In addition, kinetics of the final oocyte maturation (FOM) process, ovulation rate, and egg quality of each group was monitored (n = 12 for each group). The results indicated that there were no differences in terms of morphometry, FOM kinetics, and most stress indices between groups throughout the experiment, except haematocrit, which increased immediately following the acclimation period in fish kept in darkness. Constant darkness negatively affected plasma levels of 17α,20ß-dihydroxy-4-pregnen-3-one (DHP) and LH transcript expression at the time of the second hormone injection. This indicated that exposure to constant darkness negatively affected priming of the hormonal dose applied, resulted in the disruption of ovulation, and reduced ovulation rates (50%) for group 0L, as compared to 16L (91%). The findings of this study clearly indicate that constant darkness may have significant deleterious effects on reproductive traits throughout out-of-season induced, hormonally supported, controlled reproduction. Therefore, we advise against the use of constant darkness when managing broodstock reproduction in domesticated Eurasian perch.

Lactoferrin, lysozyme and immunoglobulin G content in milk of four breeds of cows managed under intensive production system.

The aim of the study was to analyze chosen antibacterial protein content, i.e. immunoglobulin G, lactoferrin and lysozyme in bovine milk obtained from cows of four breeds raised and maintained under intensive production technology, that is in the free stall system, and fed total mixed ration diet (TMR system). The studies were conducted on milk from four breeds of dairy cows maintained in Poland, i.e. Polish Holstein-Friesian Black-White and Red-White variety, Jersey and Simental. Milk samples were collected solely from healthy cows, having performed the TOK test with Mastirapid. Further analyses were made only on the milk samples with SCC determined under 2.0 x 10(5) cells/ml, i.e. 423 samples. Each sample was examined for lactoferrin and lysozyme determination using reversed-phase high-performance liquid chromatography (RP-HPLC) with UV-VIS detector. The immunoglobulin G (IgG) levels were established by the aid of radial immunodiffusion technique with Bovine IgG LL tests (The Binding Site, Birmingham, UK). It should be stated that a breed of cow had a significant effect on a content of the antimicrobial proteins analyzed in bovine milk evaluated. Milk obtained from Simental and Jersey cows prove to be their excellent source, i.e. lactoferrin (116.74 and 103.48 mg/l), lysozyme (9.84 and 13.02 microg/l) and immunoglobulin G (579.9 and 508.6 mg/l). The highest content of these protein was stated in milk of multiparous cows at the late stage of lactation.

The value of tumor-stroma ratio as predictor of pathologic response after neoadjuvant chemoradiotherapy in esophageal cancer.

BACKGROUND AND PURPOSE: With currently available techniques, the prediction of pathologic complete response after neoadjuvant chemoradiotherapy is insufficient. The tumor-stroma ratio (TSR) has proven to be a predictor of survival for several types of cancer, including esophageal. The aim of this study was to investigate the value of TSR in predicting pathologic response after neoadjuvant chemoradiotherapy in esophageal cancer patients. MATERIALS AND METHODS: Patients with esophageal adenocarcinoma or squamous cell carcinoma who received neoadjuvant chemoradiotherapy followed by a resection were selected. Haematoxylin and eosin (H&E) stained sections of diagnostic biopsies were collected and TSR was independently assessed by two investigators. Patients were categorized in stroma-low (≤50% stroma) and stroma-high (>50% stroma) groups for further analyses. The tumor regression grade (TRG) was assessed on H&E stained sections of the resected primary tumor to determine pathologic response. RESULTS: A total of 94 patients were included in this study, of which 76 patients were categorized as stroma-low and 18 as stroma-high. Forty-two (45%) patients had a major pathologic response (TRG 1-2), whereas 52 (55%) were considered non-responders. After adjustment for gender, tumor type, cT-status and differentiation grade, patients with a stroma-high tumor showed a higher chance of no response compared to patients with a stroma-low tumor (OR 3.57, 95%CI 1.03-12.31, P = 0.04). CONCLUSION: TSR showed to have the potential to aid in the prediction of pathologic response in esophageal cancer patients receiving neoadjuvant chemoradiotherapy. Larger validation studies are necessary before implementing this method in daily practice.

Activity of base analogues (5-fluorouracil, 5-flucytosine) against planktonic cells and mature biofilm of Candida yeast. Effect of combination with folinic acid.

BACKGROUND: The increasing number of Candida infections, especially those caused by non-C. albicans species and resistant strains, is a serious medical problem. OBJECTIVES: In this study, the antifungal activity of base analogues, 5-flucytosine (5-FC) and 5-fluorouracil (5-FU), was tested against planktonic cells as well as against mature biofilm. METHODS: Tests were performed according the EUCAST methodology. Antibiofilm effectiveness of tested drugs was determined by the crystal violet staining method. The cytotoxicity assays was performed according to the ISO 10993-5 norm. RESULTS: 5-FC and 5-FU were effective against fifteen fluconazole resistant Candida glabrata strains with an average minimal inhibitory concentration (MIC) of 0.152mg/L and 0.39mg/L, respectively. Folinic acid (folinate- e.g., leucovorin) is a common drug used in oncology simultaneously with 5-FU. In our tests folinate was able to lower MIC for 5-FC from 0.152 to 0.058mg/L (P<0.05). In the biofilm assay 5-FU and 5-FC alone did not induce any changes in the biomass of mature biofilm. Addition of folinate to each base analogue resulted in up to 90% reduction of biomass. Viability tests show that a concentration of 64mg/L of 5-FC and 5-FU supplemented with folinate can be fungicidal against mature biofilms of some Candida isolates. No cytotoxic effect was found for combination of FOL and 5-FC. CONCLUSION: Therapy of 5-FU+folinate is well known in cancer treatment, in this study we reveal the beneficial effect of folinate on antifungal activity of 5-FC as well as the antifungal potential of 5-FU+folinate.

The susceptibility of Candida albicans strains to selected anticancer drugs and flucytosine, relevance of the presence of self-splicing intron in 25S rDNA.

BACKGROUND: The presence of intron 25S allows to divide the Candida albicans species into three subclasses (A, B, C). Intronless and intron harboring strains were reported to have different susceptibility to some drugs, for example to flucytosine and bleomycin. OBJECTIVES: In this paper we tested the activity of selected antineoplastic agents, bleomycin, mitomycin C, dactinomycin and fluorouracil as well as antifungal drug flucytosine against 49 C. albicans isolates. Twenty-four strains used in this work contained intron, whereas twenty-five were intronless. METHODS: The minimal inhibitory concentrations were determined by the standard microdilution method according to EUCAST. RESULTS: All of the tested agents showed antifungal activity. Bleomycin was the strongest with an average minimal inhibitory concentration [MIC] of 15.5mg/L (range: 2-32), while the highest MIC was found for dactinomycin: 172.14mg/L (range: 128-256). Intron harboring strains seem to be more susceptible to bleomycin and flucytosine; however, differences were not statistically significant. The only two strains with elevated MICs for flucytosine were intronless. In contrast, the MIC of 5-fluorouracil was more than two times lower in intron harbouring strains comparing to intronless strains (P-value=0.0124). We found that the addition of folinate significantly increased the susceptibility of intronless strains to fluorouracil. MIC of fluorouracil decreased in this group from 58.24 (range: 16-256) to 16,78mg/L (2-64) after the supplementation of folinate. CONCLUSION: The antifungal potential of tested substances, especially the simultaneous action of fluorouracil and folinate (combination used in oncology), is encouraging further research.

Anti-candidal activity of selected analgesic drugs used alone and in combination with fluconazole, itraconazole, voriconazole, posaconazole and isavuconazole.

BACKGROUND: The antifungal activity of some popular analgesic drugs was postulated by several authors. OBJECTIVES: The aim of this study was to verify the antifungal effectiveness of acetylsalicylic acid, ibuprofen, metamizole, meloxicam and paracetamol against triazole-susceptible and triazole-resistant Candida spp isolates. METHODS: The minimal inhibitory concentration as well as fractional inhibitory concentration of selected analgesics and triazole on eighteen Candida spp isolates were determined by the standard microdilution method according to EUCAST. RESULTS: Among five tested compounds, only ibuprofen showed an antifungal effect against all tested isolates. In combination tests of analgesics with triazole, three types of interaction have been observed: indifference, synergism and antagonism. An azole-resistant strain of C. albicans demonstrated synergism with IBU and each of the 5 tested azole. Posaconazole also demonstrated synergism with IBU in two other cases, namely a C. albicans strain and with C. glabrata, which were both azole-resistant. Combination of FLU and IBU in two cases resulted in antagonism: in experiments with C. glabrata and C. tropicalis. This interaction was also observed when metamizole and fluconazole were used in combination on C. krusei. CONCLUSIONS: The results obtained by us confirmed antifungal activity of ibuprofen, what support purposefulness to continue the study on antimicrobial activity of this group of drugs.

Providencia alcalifaciens as the presumptive cause of diarrhoea in dog.

A case of diarrhoea in a four-month-old golden retriever is described. On the basis of anamnesis and bacteriological examination, the diagnosis of bacterial enteritis due to Providencia alcalifaciens was reached. Although there are contradictory opinions about the role of this organism as the enteric pathogen, it seems that Providencia alcalifaciens should also be taken into consideration in the routine bacteriological diagnostics of diarrhoea in dogs.

Comparative lipid analysis of purified plasma membranes and shed extracellular membrane vesicles from normal murine thymocytes and leukemic GRSL cells.

The lipid fluidity in purified plasma membranes (PM) of murine leukemic GRSL cells, as measured by fluorescence polarization, is much higher than in PM of normal thymocytes. This was found to be due to relatively low contents of cholesterol and sphingomyelin and a high amount of unsaturated fatty acyl chains, especially linoleic acid, in the phospholipids. PM from GRSL cells contain markedly more phosphatidylethanolamine than those from thymocytes. For both GRSL cells and thymocytes the detailed lipid composition of isolated PM was compared with that of the corresponding shed extracellular membranes (ECM), which were isolated from the ascites fluid and from thymus cell suspensions, respectively. The somewhat decreased lipid fluidity of thymocyte ECM as compared to their PM, can be ascribed to the increased cholesterol/phospholipid molar ratio (0.88 vs. 0.74). No other major differences were found between the lipid composition of these membranes. In contrast, significant differences were found between PM and ECM from GRSL cells. In this system a much lower lipid fluidity of the shed ECM was found, due to the much increased cholesterol/phospholipid molar ratio (3.5-fold) and sphingomyelin (9-fold) content, as compared to the PM. Further, the ECM contain relatively more lysophosphatidylethanolamine and less phosphatidylcholine and -inositol. ECM contain a higher amount of polyunsaturated fatty acids, especially in the phosphatidylethanolamine and lysophosphatidylethanolamine classes. On the other hand, the fatty acids of phosphatidylcholine and lysophosphatidylcholine are more saturated than in PM. In particular, ECM of GRSL cells contain less oleic and linoleic acid residues and more arachidonic acid and 22:polyunsaturated fatty acid residues than PM. The possible relevance of these differences with respect to the mechanism of shedding of vesicles from the cell surface, is discussed.

Studies on plasma membranes. XXII. Fatty acid profiles of lipid classes in plasma membranes of rat and mouse livers and hepatomas.

Plasma membranes were isolated from rat and mouse livers, a transplanted rat hepatoma, two transplanted mouse hepatomas and spontaneous mouse hepatomas. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, lysophosphatidylcholine, free fatty acids and triglycerides were separated and their fatty acid profiles determined. The various lipid classes of rat and mouse liver plasma membranes each demonstrated more or less specific fatty acid profiles. The number of double bonds decreased in the order: phosphatidylserine greater than or equal to phosphatidylethanolamine greater than phosphatidylinositol greater than phosphatidylcholine greater than or equal to sphingomyelin and lysophosphatidylcholine. Small species differences were noted in most lipid classes. A marked sex difference was observed in sphingomyelin of mouse liver membranes but in none of the phospholipids of rat liver membranes. The increased cholesterol content of all hepatoma versus liver plasma membranes was accompanied by a decrease of fatty acyl poly-unsaturation in most lipid classes of the rat hepatoma but not of the mouse hepatoma membranes. The fatty acid profiles of the mouse hepatoma membranes deviated much less from those of mouse liver than did the pattern of rat hepatoma versus rat liver. The results were discussed in relation to lipid fluidity of which fatty acyl unsaturation and cholesterol are the main parameters.

Molecular characterization of pssCDE genes of Rhizobium leguminosarum bv. trifolii strain TA1: pssD mutant is affected in exopolysaccharide synthesis and endocytosis of bacteria.

We have identified the three genes pssCDE in Rhizobium leguminosarum bv. trifolii TA1. Even though they were almost identical to earlier identified pssCDE genes of R. leguminosarum, they differed in gene lengths and gene overlaps. The predicted gene products of pssCDE genes shared significant homology to prokaryotic glycosyl transferases involved in exopolysaccharide synthesis. The Tn5 insertion in pssD created the nonmucoid mutant that induced non-nitrogen-fixing nodules. The microscopic analysis of the nodules, induced on Trifolium pratense by the pssD133 mutant, showed abnormally enlarged infection threads densely packed with bacteria, which were released from the infection threads in an unusual way. The symbiosomes were observed very rarely and the nodule remained almost empty. Symbiotic phenotype of the pssD133 suggested a correlation between this mutation and defective endocytosis of bacteria into nodule cells.

Rapidly progressive septic shock, associated with necrotizing fasciitis.

A case is presented of a 59-year-old male with rapidly progressive septic shock and necrotizing fasciitis. The patient was admitted in shock with an extensive skin lesion on the anterior chest wall. The history was relatively short and there was only a questionable history of preceding trauma. Necrotizing fasciitis was suspected from the appearance of the lesion. Antibiotics and anti-shock therapy were given but despite this, his condition deteriorated and he died from septic shock. At autopsy, the diagnosis of necrotizing fasciitis was confirmed. The importance of rapid diagnosis and primary surgical therapy is emphasized.

The pssB gene product of Rhizobium leguminosarum bv. trifolii is homologous to a family of inositol monophosphatases.

The Rhizobium leguminosarum bv. trifolii region encoding pssA and pssB genes was cloned. The pssB gene located upstream of the pssA encoded a 28.36-kDa protein which displayed 97.5% identity with the PssB of R. leguminosarum bv. viciae. Inactivation of the pssB gene by insertion of the lacZ-Gmr cassette resulted in the significant increased production of exopolysaccharide in comparison to the wild-type level. A mutant strain was also defective in nitrogen fixation suggesting a regulatory role of pssB in symbiosis with clover.

Effect of superior colliculus lesions on sensory unit responses in the intralaminar thalamus of the rat.

The effects of kainic acid lesions of the intermediate and deep layers of the superior colliculus on the sensory input to the intralaminar thalamus of the rat were determined. Ipsiversive circling and contralateral sensory neglect were consistently seen after lesion placement. Two to 7 days later, the intralaminar thalamus was systematically explored for extracellular mechanoreceptive unit responses to high threshold and low threshold stimuli. On the side ipsilateral to the lesion the number of responsive units was reduced by 51%. The loss was particularly marked for nociceptive units (80%), and low threshold and complex units with orofacial receptive fields (73%). This effect may involve a partial deafferentation of the intralaminar thalamus as well as altered excitatory thresholds of thalamic neurons. It is suggested that the functionally distinct direct tectothalamic projection as well as the indirect tecto-reticulo-thalamic pathway are implicated.

Molecular characterization and symbiotic importance of prsD gene of Rhizobium leguminosarum bv. trifolii TA1.

The prsD, prsE and orf3 genes of Rhizobium leguminosarum bv. trifolii strain TA1 encode the proteins which are significantly related to the family of bacterial ABC transporters type I secretion systems. The prsD:Km(r) mutant of strain TA1 induced non-nitrogen-fixing nodules on Trifolium pratense. Microscopic analysis of the nodules induced by prsD mutant did not reveal major abberations in the bacteroid appearance. The exopolysaccharide of prsD mutant was produced in increased amount and its level of polymerization was changed. SDS/PAGE of the proteins from the culture supernatants showed a lack of the 47-kDa protein in the culture of prsD mutant. Thus, PrsD may play a role in the export of this protein.