Quantitative characterization of trace levels of PFOS and PFOA in the Tennessee River.

Although there is evidence of widespread distribution of organic fluorochemicals such as perfluorooctane sulfonate and perfluorooctanoate, in the environment, the versatility of these compounds in industrial and commercial applications complicates characterization of pathways into the environment. A solid-phase extraction method coupled with HPLC-negative-ion electrospray tandem mass spectrometry was developed to quantitatively measure trace levels of organic fluorochemicals in drinking water and surface water. Using this method, certain fluorochemicals can be quantitatively measured in water samples down to 25 ppt, a level well below calculated drinking water advisory levels. To assess fluorochemical distribution in a localized geography and to ascertain whether fluorochemical manufacturing facilities contribute to environmental levels of fluorochemicals, 40 water samples were collected on an 80-mi stretch of the Tennessee River, near a fluorochemical manufacturing site in Decatur, AL. Low levels (ppt) of perfluorooctane sulfonate were determined throughout the stretch of river sampled. Concentrations of the measured fluorochemicals increased downstream of the fluorochemical manufacturing facility, indicating that effluent from manufacturing is one likely source of organic fluorochemicals into the river.

UV resonance Raman spectra reveal a structural basis for diminished proton and CO2 binding to alpha,alpha-cross-linked hemoglobin.

UV resonance Raman difference spectra between ligated and deoxyhemoglobin contain tryptophan and tyrosine signals which arise from quaternary H-bonds in the T state, which are broken in the R state. These H-bonds are unaffected by bis(3,5-dibromosalicyl) fumarate cross-linking at the Lys alpha 99 residues, which prevents dissociation of Hb tetramers to dimers. However, when the pH is lowered from 9.0, or when NaCl is added, intensity is diminished for the tyrosine Y8 and tryptophan W3 bands of cross-linked deoxyHb, but not of native deoxyHb. This effect is attributed to weakening of tertiary H-bonds involving Tyr alpha 140 and Trp alpha 14, when the T state salt bridge between Val alpha 1 and Arg alpha 141 is formed via protonation of the terminal amino group and anion binding. The Tyr alpha 140-Val alpha 93 H-bond connects the Arg alpha 141-bearing H helix with the Lys alpha 99-bearing G helix. Weakening of the H-bond reflects a tension between the fumarate linker and the salt-bridge. This tension inhibits protonation of the Val alpha 1 amino terminus, thus accounting for the diminution of both proton [Bohr effect] and CO2 binding in the T state as a result of cross-linking.

Fourier transform infrared evidence against Asp beta 99 protonation in hemoglobin: nature of the Tyr alpha 42-Asp beta 99 quaternary H-bond.

The Tyr alpha 42-Asp beta 99 intersubunit H-bond stabilizes the T quaternary structure in hemoglobin (Hb) tetramers. We had proposed that Tyr alpha 42 acts as an acceptor in this H-bond, because the tyrosine Y8a/8b and Y7a' UVRR (ultraviolet resonance Raman) bands shift in directions opposite to those expected if tyrosine is an H-bond donor. If Asp beta 99 is the H-bond donor, then it must be protonated in the T state, and would be a previously unrecognized contributor to the Bohr effect. This implication was strengthened by the discovery that an R-minus-T difference FTIR (Fourier transform infrared) band at 1693 cm-1, which might be a signal from protonated carboxylate, is missing in Hb Kempsey, a mutant in which Asp beta 99 is replaced by Asn. However, we now find that this FTIR signal is insensitive to 13C-labeling of the aspartate residues in Hb, and cannot arise from protonated Asp beta 99. There are no other difference signals in the 1700 cm-1 region at a sensitivity of one COOH group. We conclude that Asp beta 99 is not protonated, and that the anomalous UVRR shifts must arise from compensating polarization of the Tyr alpha 42 OH. Candidates for this compensation are the H-bond donated by the Asp beta 94 backbone NH, and the nearby positive charge of Arg beta 40.

Haematology and histopathology of seven-week-old broilers after early food restriction.

The haematology and histopathology of seven-week-old broilers were examined after periods of early food restriction, for six, 10 or 14 days from six days old. After several weeks on an ad libitum diet the birds failed to compensate for the weight lost during early food restriction. Immediately after the periods of food restriction, the birds demonstrated significantly increased heterophil/lymphocyte ratios, reduced eosinophils and slightly raised basophil counts. At seven weeks old, a significant reduction was seen in red and white blood cells and thrombocyte numbers together with significant increases in mean cell haemoglobin and mean cell volume. The haematological profile demonstrated a macrocytic normochromic anaemia caused possibly by a folic acid deficiency as a result of the food restriction. Histopathological lesions were seen in the heart, lungs and liver from birds on all diets but there were more lesions the longer the food had been restricted. Lung disease was more marked where there was inadequate ventilation. Cartilaginous and osseous lung nodules were significantly fewer after food restriction. It was postulated that the increase in pathological lesions in the food-restricted birds may be associated with a stress response.

Ectopic cartilaginous and osseous lung nodules induced in the young broiler by inadequate ventilation.

The study attempted to induce cartilaginous and osseous nodules in the lungs of young broilers by providing them with inadequate ventilation. In experimental birds there were more nodules than in two groups of control birds, with significantly more occurring in the left lung than in the right. A majority of the nodules were fibrous. Histologically, the lungs of the experimental birds had lesions similar to those seen in an ascites syndrome or with induced hypoxia. In another experiment, when the lungs of healthy adult broiler breeders were examined, more osseous-type nodules were present. Very few nodules were seen in the lungs of wild birds. It was suggested that poor ventilation, similar to that likely to be experienced in some bird accommodation, may increase the development of cartilaginous and osseous nodules in the lungs of broiler chickens.

The incidence of ectopic cartilaginous and osseous lung nodules in young broiler fowls with ascites and various other diseases.

The incidence of ectopic cartilaginous and osseous lung nodules in young broiler fowls with ascites and various other diseases has been determined. In all groups of diseased birds there were more nodules in the lungs compared with control birds. The birds that died with an ascitic syndrome had a significantly greater number of nodules than the other groups of diseased birds, with significantly more occurring in the left lung than in the right (P<0.001). Histologically, the fibrous cartilaginous type of nodule was the most numerous and the osseous type the least numerous. This is the first detailed study in fowls in which a relationship has been demonstrated between the presence of cartilaginous and osseous lung nodules and various disease states.

Effect of gamma-radiation on fowl sperm function in vitro and in vivo.

Doses of up to 300 Gy of ionizing radiation had little effect on fowl sperm morphology, ATP content and motility when measured in vitro. Fertility of eggs from hens inseminated with spermatozoa receiving 50 Gy, in terms of post-oviducal development, was less than 4%. However, 35% appeared 'fertile' by macroscopic examination of the germinal disc of unincubated eggs. These contained few embryonic cells, although the vitelline membrane contained many trapped spermatozoa. After doses of 100 Gy or more, inseminated spermatozoa were not found in the vitelline membrane and no fertile or apparently fertile eggs were produced; nor did such spermatozoa enter the utero-vaginal sperm-host glands. Genetic transformation using fowl spermatozoa irradiated with doses in excess of 100 Gy appears to be an unlikely prospect.